WO2025079065A1 - Fertilizer production using nitrogen fixing cyanobacteria - Google Patents

Abstract

A method and system produce fertilizer products by growing cyanobacteria in a photobioreactor, the cyanobacteria performing nitrogen fixation using Nitrogen from the ambient air brought into the photobioreactor. The grown cyanobacteria is harvested from the photobioreactor as biomass, which is subject to anaerobic digestion in an anaerobic digestion reactor, to produce Ammonium, which is optionally subjected to nitrification, in a nitrification bioreactor, to produce the resultant fertilizer product.

Description

Fertilizer Production Using Nitrogen Fixing Cyanobacteria

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is related to and claims priority from commonly owned US Provisional Patent Application Serial No. 63/588,724, entitled: Fertilizer Production Using Nitrogen Fixing Cyanobacteria, filed on October 8, 2023, which is related to commonly owned patent application WO 2020/044279 Al (PCT/IB2019/057284), entitled: Process For Biological Ammonia Production By Nitrogen Fixing Cyanobacteria, the disclosures of both patent applications are incorporated by reference herein in their entirety.

TECHNICAL FIELD

This application is directed to methods for producing nitrogen-based fertilizers.

BACKGROUND

Nitrogen fertilizer is a major pillar in securing food, as it supports highly efficient intensive agriculture. The invention of synthetic ammonia production from nitrogen and hydrogen from water, by Haber and Bosch was a major breakthrough over 100 years ago, and as of today, is the leading practical way to feed the world's population. However, the process is energy intensive, causing large amounts of CO₂ emissions, and is not considered organic.

Furthermore, organic farm products are gaining growing importance providing healthier, and less polluting alternatives to intensive classical agriculture. Modern organic precision agriculture uses fertigation such as hydroponics, drip irrigation, and sprinklers, and depends on steady supplies of high quality available nitrogen rich liquid fertilizer, often not achieved by current products that are based on extracts from waste biomass and can encompass undesired contaminations such as hormones, pharmaceuticals, antidepression drugs, or antibiotic residues, and offer often variable and poorly exploitable available nitrogen, the available nitrogen being Ammonia or Nitrate, as opposed to unavailable nitrogen, which is found in most current products that are based on extracts from waste.

SUMMARY OF THE DISCLOSURE

The disclosed subject matter is directed to a method for producing atmospheric nitrogen-based fertilizers. The method comprises: obtaining hydrogen from water; sourcing Nitrogen and Oxygen from ambient air; sourcing Carbon Dioxide from a gas source and/or from the air, and, feeding the water, ambient nitrogen and Oxygen, and Carbon Dioxide, and typically media, such as nutrients, into a bioreactor, such as a photobioreactor, the bioreactor populated with an effective amount of one or more strains of: bacteria, for example, cyanobacteria, also sometimes referred to as microalgae, and/or microorganisms, to maintain stability in the bioreactor, to facilitate photosynthesis, that performs carbon capture, and nitrogen fixation by the cyanobacteria, to grow biomass, which includes the cyanobacteria, for the production of Biofertilizer and/or protein-rich food.

BRIEF DESCRIPTION OF THE DRAWINGS

Non-limiting examples of embodiments are described below with reference to figures attached hereto that are listed following this paragraph. Identical structures, elements or parts that appear in more than one figure are generally labeled with the same numeral in all the figures in which they appear. Dimensions of components and features shown in the figures are chosen for convenience and clarity of presentation and are not necessarily shown to scale.

In the drawings:

FIG. 1 is a block diagram of the disclosed system in an example operation;

FIG. 2A is a block diagram of the Photobioreactor System of FIG. 1;

FIG. 2B is a block diagram of the Fertilizer System of FIG. 1;

FIG. 2C is a block diagram for the controller of FIGs. 1, 2A and 2B;

FIG. 3 is a diagram of an example Photobioreactor of FIG. 2A;

FIG. 4 is another diagram of a system of the Photobioreactor of FIG. 2A;

FIG. 5 is a block diagram of the Cleaning In Place System of FIG. 2A;

FIG. 6 is a screen shot of a diagram of the control system for the Photobioreactor System disclosed herein;

FIG. 7 is an overview screen associated with the screen shot of FIG. 6

FIG. 8 is a diagram of the hierarchy of the modules of the control system for the Photobioreactor System;

FIG. 9 is screen shot showing triggers;

FIG. 10 is a diagram of the relationship between various components of the software for running the control electronics of FIG. 6 FIGs. HA and 11B are a flow diagram of an example process performed by the Photobioreactor System disclosed herein; and

FIG. 12 is a flow diagram of an example process performed by the Fertilizer System disclosed herein.

DETAILED DESCRIPTION OF THE DISCLOSED SUBJECT MATTER

The present disclosure provides for methods for producing nitrogen-based fertilizers with the nitrogen source being from ambient air.

The present disclosure provides a process for manufacturing fertilizer, that has sufficient quantities of available Nitrogen in the form of Ammonium (NH4) and/or Nitrates (NO₂). The process involves the production of biomass, from a culture of cyanobacteria (e.g., cyanobacteria cells), minerals, gases, and water, provided with energy from sunlight, to grow the cyanobacteria, e.g., cyanobacteria culture, in a photobioreactor (PBR). The “biomass” as referenced, used, and described, herein, is the product produced in, and harvested from, the photobioreactor, and includes, at least cyanobacteria, and nutrients, in a liquid, e.g., water, solution. The cyanobacteria culture may be introduced with an inhibitor. When this is the case, the harvested biomass will also include Ammonium (NH4) released from the cyanobacteria cells and dissolved in the medium.

Alternatively, by changing the pH (of the liquid, e.g., water, solution/the culture), the Ammonium (NH4) converts to Ammonia (NH3) which is released as gas from the medium and can be captured using an Ammonia trap.

Initially, cyanobacteria, for example, a cyanobacteria culture (of cyanobacteria cells filaments including heterocysts (heterocyst cells) and other cells, which are non -heterocysts (nonheterocyst cells), are grown in a photobioreactor (PBR) or bioreactor. The terms, “PBR”, “photobioreactor”, and “bioreactor”, are used interchangeably herein. The cyanobacteria, e.g., cyanobacteria cells, which are heterocysts (heterocyst cells) perform nitrogen fixation (the process of converting nitrogen gas (N2) in its heterocyst cells to Ammonia (NH3) and/or Ammonium (NH4) as it grows in the photobioreactor.

At the same time, other cyanobacteria cells (e.g., the non-heterocyst cells) perform photosynthesis, by capturing carbon, and converting the sun's energy (e.g., sunlight) into chemical energy, such as sugar, which is used by the heterocysts as its energy source for the nitrogen-fixing and the making of Ammonia and/or Ammonium.

The cyanobacteria grown in the photobioreactor, is harvested as biomass (e.g., wet biomass), using for example a centrifuge as a harvesting unit or harvester. The harvested biomass is then subjected to anaerobic digestion (AD), in an anaerobic digester or Anaerobic Digestion Bioreactor. In the anaerobic digester, the cyanobacteria of the biomass is converted to Ammonium (NH4) by anaerobic digestion with anaerobic bacteria, resulting in Methane Gas (CH4) and an Ammonium (NH4) rich nutrient effluent. Next, a Nitrification Process in a Nitrification Bioreactor occurs as the Ammonium of the nutrient effluent mixes with, for example, Calcium Carbonate or other carbonate, and, for example Magnesium or Potassium. Aerobic bacteria in the Nitrification Bioreactor convert the Ammonium (NH4) into Nitrates (NO₂), such as Nitrate -based salts, such as Calcium Nitrate, Potassium Nitrate, or Magnesium Nitrate, which are high-value fertilizers.

The present disclosure provides methods and systems for producing fertilizer products by growing cyanobacteria in a photobioreactor, the cyanobacteria performing nitrogen fixation using Nitrogen from the ambient air brought into the photobioreactor. The grown cyanobacteria is harvested from the photobioreactor as biomass, which is subject to anaerobic digestion in an anaerobic digestion reactor, to produce Ammonium, which is optionally subjected to nitrification, in a nitrification bioreactor, to produce the resultant fertilizer product.

The present disclosure also provides processes and systems for the production of liquid nitrogen rich organic fertilizer by photosynthetic production of nitrogen fixing cyanobacteria in biomass, the cyanobacteria, for example, of the genus Anabaena or other nitrogen fixing cyanobacteria in photobioreactors (PBRs). The disclosed process performed in the photobioreactor (PBR) to refine the cyanobacteria cells, for example, in a culture, which when provided energy from sunlight activated photosynthesis, perform nitrogen fixation, a biological process, reliant on sunlight or light and cyanobacteria and, for example, also optionally, inhibitors.

The photobioreactor process is followed by anaerobic digestion (fermentation) of the resulting biomass in an Up Flow Anaerobic Sludge Blanket (UASB) or other type of anaerobic digester (AD) or anaerobic digestion bioreactor. The anaerobic digestion in the anaerobic digestion bioreactor yields both biogas, that is being used for electric and heat energy generation, and a steady flow of nutrient rich Ammonium effluent (nutrient effluent) suitable for use, for example, as liquid organic fertilizer in hydroponics, dripirrigation, or other precision agriculture.

The process employs photobioreactor (PBR) systems, for example, having airlift columns or other similar design, geared to optimal performance via appropriate sensors and control technology, adapted to climate variations by proprietary algorithms and machine learning to control growth medium regeneration and supply, CO₂ supply, temperature, and harvesting volumes. The process allows production of renewable nitrogen fertilizer, with Nitrogen captured from ambient air by cyanobacteria, in a circular zero emission Green House Gases (GHG) neutral, virtually emission free process. Biomass is harvested using a centrifuge and/or an advanced microfiltration cross-membrane device (for example, from SANI Membranes A/S of Denmark) and concentrated (dewatered), for example, approximately 20 to 40 fold, so as to be suitable for feeding the Anaerobic Digestion (AD) process.

Harvested biomass is fed into an anaerobic digester or anaerobic digestion bioreactor, operated, for example, as an anaerobic membrane bioreactor, as an Up Flow Anaerobic Sludge Blanket (Reactor) (UASB), or in an Anaerobic Digestion (AD) mode. Resulting biogas is combusted for electricity generation, with both electricity and resulting heat being used to improve process sustainability. Air or Carbon Dioxide (CO₂) rich air is fed into the photobioreactor. Resulting Ammonium in a nutrient rich effluent (nutrient effluent) is sterile filtered to be used as organic liquid fertilizer. The nutrient effluent, for example, with approximately 2000 to 2500 ppm nitrogen content, can be applied directly to hydroponics or drip irrigation in greenhouses with the Photobioreactor (PBR) system, for example, as a standalone addition to the greenhouse technology, shown as a rotating field system for growing plants, as disclosed in, for example, commonly owned US Patent No. 10,980,198, entitled: Automatic Hydroponic Greenhouse Factory, the disclosure of which is incorporated by reference in its entirety herein.

In a second embodiment, the nutrient effluent (from Anaerobic Digestion) can be concentrated using reverse osmosis to reach up to 50 gram Nitrogen/liter (N/l). The fertilizer composition is, for example, adjusted by optimizing micro- or oligo-elements concentrationsusing dissolved reactor precipitate according to analysis results, to provide for a high quality organic liquid fertilizer formulation, as disclosed herein.

FIG. 1 shows the disclosed system 100 for cyanobacteria growth in biomass and fertilizer production, in an example operation with a table of a rotating field system 102, such as that of US Patent No. 10,980,198. The system includes a biomass Photobioreactor (PBR) System 110 and a Fertilizer System 120. The Fertilizer System 120 feeds the rotating field system 102, for example, by feeding fertilizer into the liquid solution of the water bed of the rotating field table, which holds trays with the growing plants in a circulation in the liquid solution of the rotating field table.

The system 100 includes the Photobioreactor (PBR) System 110 and the Fertilizer System 120. The PBR system 110 and Fertilizer System 120 may be together or separate and both systems 110, 120 may be automatically controlled by a single controller 130 (FIG. 1), or by separate controllers 140, 141 (FIGs. 2A and 2B), the controllers 130, 140, 141 all including a computer 150 (FIG. 2C). The photobioreactor system 110 and fertilizer system 120 may also be manually controlled, or manual control in combination with the controllers 130, 140, 141.

When together, the Photobioreactor System 110 and Fertilizer System 120 may be in fluid communication with each other, connected by lines, through which product (e.g., biomass, such as wet biomass including bacteria cultures grown in a Photobioreactor 200 of the system 110) can be pumped (by pumps valves and the like) or otherwise automatically transferred, from components of the Photobioreactor System 110 to the Fertilizer System 120 (e.g., the Anaerobic Digestion Bioreactor 300). The components of both systems 110, 120 including components for transferring product from the Photobioreactor System 110 to the Fertilizer System 120 are, for example controlled by one or more of the controllers 130, 140, 141.

As shown in FIG. 2C, the controllers 130, 140, 141 may include one or more processors, for example, in the form of a Central Processing Unit (CPU) 150 (so as to include a “computer”) and one or more memories 154 (some or all with associated storage), for example, which store machine executable instructions for execution by the CPU 150, for executing the processes, computer programs, algorithms, and the like disclosed herein and detailed below, to operate the components, models, and the like, disclosed herein. The processors of the CPU 150 execute various programs stored in the memory 154, and process various data in the various programs, and other functions, of the system 100 and/or systems 110, 120, as disclosed herein. The controllers 130, 140, 141, for example, control the pumps (P) 160, the valves (V) 161, the sensors 162 (detailed below), other functionalities 163 and the manifold components 164, including, for example, pumps, valves, sensors, bubblers/gas injectors (for airlift in the columns 352), taps, drains, heaters and coolers, and the like.

The memory 154 may store operation data of various programs for operation of the respective controller 130, 140, 141 and may be in single or plural units as needed. The memory may be volatile memory or non-volatile memory. Volatile memory may include RAM, DRAM, SRAM, and the like, while non-volatile memory may include ROM, PROM, EAROM, EPROM, EEPROM, Flash Memory, and the like. The memory 154 may also store one or more machine learning (ML) models 156, which typically include trained models, with data added to the training data upon each triggering event. This allows models to become familiarized with their specific location, and thus, can adjust for various local climate conditions, local times, and seasons.

For example, a machine learning (ML) model 156, for triggering discharge, for example, in determined amounts, for the photobioreactor 200, e.g., photobioreactor columns 352, for example, for harvesting by the harvesting unit (HU) 270, depends on biomass density, which is typically determined based any combination of parameters, such as light, sensed by light sensors, colors, sensed by color sensors, images from cameras, pH (by a pH sensor), temperature (by temperature sensors, such as thermometers), ambient humidity (by a humidity sensor), ambient air pressure (by a pressure sensor including a barometer), and, air quality (by an air quality sensor). These parameters are also usable in a computer program for harvesting, times and amounts to harvest.

The Photobioreactor System 110 includes a photobioreactor (PBR) 200, as a bioreactor, where cyanobacteria, e.g., cyanobacteria cultures of cyanobacteria cells, are grown. The photobioreactor 200, is for example, operative with light, such as natural sunlight 202, or other light sources, the light which provides the energy to drive the disclosed process of photosynthesis, which provides energy for the nitrogen fixation of ambient Nitrogen in the photobioreactor 200 by the cyanobacteria cells, in the photobioreactor 200. The photobioreactor 200 is formed, for example, of one or more columns 352, as arranged in one or more rows, as detailed below, and shown, for example, in FIG. 3. The columns 352 are, for example, transparent and/or translucent to light, such as sunlight 202, and are where gas exchange occurs between Nitrogen (N₂) gas, from the ambient air, e.g., atmospheric Nitrogen, Caron Dioxide (CO₂) gas from the ambient or from a CO₂ emitting source, and Oxygen (O₂) gas, also from the ambient air or an Oxygen source, such as or similar to, an anaerobic digester, and Hydrogen (H₂) from the water in a liquid solution (medium). These gases feed the cyanobacteria, i.e., cyanobacteria cells (e.g., heterocyst and the non-heterocyst cells) in the medium, resulting in Nitrogen fixation in the cyanobacteria heterocyst cells and photosynthesis in the other filaments cells (non-heterocyst cells). Inhibitors may optionally be applied into the respective columns 352, and if so, Ammonium (NH₄) will be released from the cyanobacteria cells in the column 352.

The cyanobacteria, i.e., cyanobacteria culture, includes bacteria, typically in the form of a heterocyst or heterocyst cell, the heterocyst cells having the ability to fix Nitrogen (N2), and microalgae. The cyanobacteria culture, for example, includes cyanobacteria from Anabaena, and, for example, includes Anabaena flos aqua, Anabaena siamensis, Anabaena azollae, Anabaena variabilis, and mutant strains thereof, the Anabaena a genie of the family Nostocacae,

The cyanobacteria is in a liquid, e.g., water, solution, in an effective amount, in the photobioreactor 200. In the photobioreactor 200, there are typically nutrients, macro and micro nutrients, and optionally inhibitors. Sunlight provides energy to the bacteria, i.e., cyanobacteria, and the algae (e.g., microalgae), which activates photosynthesis in the photobioreactor 200 producing sugar (e.g., C₆H₁₂O₆,) from the Carbon Dioxide gas (in an effective amount, brought into the photobioreactor 200), Oxygen gas (in an effective amount, from the ambient air brought into the photobioreactor 200), and Hydrogen, in an effective amount, from the water of the liquid solution. The sugar provides the energy for the cyanobacteria heterocysts, i.e., cyanobacteria heterocyst cells, to fix Nitrogen (from an effective amount of Nitrogen (N2) in the ambient air) brought into the photobioreactor 200 and make Ammonium (NH4) in the cells. The now grown cyanobacteria culture is harvested, with the nutrients in the waster solution, from the photobioreactor 200 as biomass (e.g., wet biomass). The cyanobacteria in the biomass is, for example, in concentrations about 0.5 to 2 milligrams/liter, with a density of approximately 1-2 grams/liter.

Electricity for operating the components of the photobioreactor system 110 and the fertilizer system 120 is generated from a Natural Gas source 210, which feeds natural gas into an energy generator 212. A photovoltaic (PV) system 214 generates additional energy, and is, for example, a supplemental system to the energy generator 210. The generated energy is sent to an Electricity Microgrid, on site or otherwise local with the system 100 or each of the systems 110, 120, to power all of the components, including all pumps and valves used by the systems 110, 120, as detailed below.

Carbon Dioxide (CO₂), from a source 220, such as a tank, and controlled by a valve (V) 221a along delivery line 221b feeds into the photobioreactor 200. For example, the Carbon Dioxide (CO₂) may be used for growing the cyanobacteria in the photobioreactor 200, while controlling pH in the photobioreactor 200, by controlling CO₂ injections with the valve 221a. For example, the Carbon Dioxide flow into the photobioreactor 200 may be controlled, for example, in a gas stream, to pressures of approximately 0.25 bar, or any other sufficient pressure to cause bubbling (e.g., airlift) of the liquid in the photobioreactor 200, sufficient for agitation (also, for example, stirring) of the liquid in the photobioreactor columns 352.

Ambient air, or treated air, which includes nitrogen (N2) and typically Oxygen (O₂), and may also be enriched with Nitrogen and/or Oxygen, is fed into the photobioreactor 200, from an intake 222, such as a manifold, over a line 225b. The line 225b and the feed rate over the line 225b, may be valve/pump (V/P) 225a controlled, so as to be controlled by a computer and/or controller 130, 140.

Water, from a tank 226 or other water source, is fed into an optional treatment unit (TMT) 228 via a pump (P) 229a, along a line 229b. The treatment unit 228 may be one or more of an ultra violet (UV) light apparatus, a reverse osmosis (RO) apparatus, and the like. From the treatment unit 228, the treated water is sent to a medium storage tank 230, via a pump (P) 231b along a line 231a. Water from the tank 226, if the treatment unit 228 is not present, is pumped via the pump (P) 229a over line 229x for cleaning by the Cleaning In Place (CIP) system 262. Should the treatment unit 228 be present, water from the treatment unit 228 is pumped by the pump (P) 231b over line 23 lx (which merges into line 229x), so that water is delivered to the CIP System 262.

Micro and macro nutrients 240, such as macro nutrients, for example, Magnesium, Potassium, Calcium, and the like, and micronutrients, for example, Boron, Copper, Sodium, Phosphorous, and the like, may be pumped from a source 240, via the pump (P) 241a and over line 241b into the medium storage tank 230.

An auxiliary (Aux.) source 256 may provide optional inhibitors, for example, inhibitors, such as MSX (L-methionine-DL-sulfoximine), MSO (L-methionine-sulfone), phosphinothricin ((RS)-2-Amino-4-(hydroxy(methyl)phosphonoyl)butanoic acid), or, Bialaphos (L-Alanyl-L- alanyl-phosphinothricin) or Glyphosate (N-(phosphonomethyl)glycine). The auxiliary source 256 may also provide optional growth medium, including, for example, Gibco® BG-11 (from Thermo-Fisher Scientific), a blue green algae medium, or a nitrogen free blue green algae medium. The components from the auxiliary source 256, may be pumped by a pump (P) 257a into the medium storage tank 230 over a line 257b, or directly into the PBR 200, via the line 257c.

Medium from the medium storage tank 230, in a liquid, e.g., water, solution, is pumped via a pump (P) 233a over a line 233b into the photobioreactor 200.

An Inoculum Reactor 250, fed with inoculum charge material from an inoculum charge source 252, such as a source of cyanobacteria (e.g., in a culture of cells), for example, as a starter culture, in accordance with the PBR Startup procedure detailed below, provides initial cyanobacteria to the photobioreactor 200. The inoculum reactor 250 may be, for example, nursery columns or other reactors, using an electric nursery valve (not shown) and dosing pumps (not shown), and function to inject the cyanobacterial and other starter cultures, and other components, such as algae, microalgae inoculum, into the photobioreactor 200. The photobioreactor 200 typically already has cyanobacteria therein, as it is initially or previously populated with cyanobacteria. Optionally the cyanobacteria can be grown in suspension on carriers, such as foams, fibers, or other materials capable of holding the cyanobacteria in place, or it receives regenerated biomass with cyanobacteria from the harvesting unit 270 (pumped in by a pump (P) 268a over line 268b), as detailed below in FIG. 4.

Hypochlorite, or other cleaning solution, from a source 260 is pumped into a Cleaning In Place (CIP) system 262, via a pump (P) 263a, and over a line 263b, to clean columns 352, tanks 404, 514, 516, and the like. Water, from the water tank 226 or treatment unit 228 is pumped (via the respective pumps (P) 229a, 231b) into the CIP system 262 as well, via line 229x (should the treatment unit 228) not be present, over line 23 lx (if the treatment unit 228 is present). The CIP system 262 is in fluid communication with the photobioreactor 200 (shown by double headed arrow 265) for cleaning and/or maintenance, as described below in FIG. 5. The CIP system 262 also removes waste water.

From the photobioreactor 200, biomass is pumped, via a pump (P) 267a over line 267b, or otherwise sent to a harvesting unit (HU) 270. For example, the process of cyanobacteria growth in the photobioreactor 200 (for example, in cycles) and its harvesting, is a continuous process of potentially an infinite number of cycles.

The biomass is, for example, such that the cyanobacteria has a density of about 1-2 grams/liter, in the medium prior to harvesting. The harvesting unit 270 may be, for example, a centrifuge (e.g., which provides up to approximately 20x concentrated biomass) or an ultrafiltration unit, such as an ultrafiltration membrane unit (for example, a VIBRO system from SANI Membranes A/S of Denmark), or a cross-membrane system. The harvesting unit 270 functions, for example, to filter or dewater or reduce density and/or concentrate the culture (for example, reducing cyanobacteria density in the culture from approximately 2 grams/liter to approximately 0.5 - 1 gram/liter), The biomass extracted is an algal slurry (e.g., a liquid) including cyanobacteria biomass (which includes protein, about 52% protein), and remove cyanobacteria biomass from the medium (liquid). The filtered culture, after the cyanobacteria biomass has been extracted from it, may then be sent back to the photobioreactor 200 over the line 268b (also described below and shown in FIG. 4), as regenerated medium. The biomass from the photobioreactor 200 is, for example, periodically harvested, for example, in accordance with a computer program or a machine learning model algorithm.

From the harvesting unit 270, the now harvested or dewatered biomass, which also includes cyanobacteria, and optionally, Ammonium in solution, if an inhibitor was in the photobioreactor 200 when the cyanobacteria was grown, for example, an algae concentrate, is sent, via a pump (P) 271a over a line 27 lb as wet biomass 274, typically for further processing. This wet biomass includes, for example, protein, i.e., the cyanobacteria, which makes the wet biomass Nitrogen rich.

Optionally, the harvested or dewatered biomass may be further treated by being pumped, via a pump (P) 275a over a line 275b into a biomass drying system 278. Biomass dryers include, for example, air dryers or freeze dryers. The same content as the wet biomass without 80% to 95% of the water. The biomass drying system 278 produces dry biomass 279, for example, for packaging and/or offsite use. The wet biomass 274 or dry biomass 279 is suitable for further processing into fertilizer, such as Nitrate (NO ), and salts thereof, such as Calcium Nitrate (Ca(NOs)2), by the Fertilizer system 120.

The photobioreactor system 110 and the fertilizer system 120 typically include sensors 162 at various locations therein, to detect conditions such as flow, volume, temperature, mass, color, pH, light, brightness, and including cameras, ambient pressure, ambient humidity, ambient air quality, feedback indicators, and the like, as well as heaters/coolers, drains, taps, timers, triggers (e.g., computer modules for triggering events), and the like, collectively element 163, may be placed with the components, along the lines and at other locations, and in communication with the controllers 130, 140, 141, to control the processes performed by the photobioreactor system 110 and the fertilizer system 120. All of the aforementioned components may be powered by energy from the electricity microgrid of the System(s) 100, 110, 120.

In the system 110, the energy used to power the components, controllers 130, 140, 141, pumps (P), valves (V), taps, sensors, heaters/coolers, drains, timers and computer modules for triggering events, is from the energy microgrid.

The wet or dry biomass, typically wet biomass, may now enter the fertilizer system 120. Initially, the dry biomass is sent, pumped, or otherwise placed into an anaerobic digester (AD) 300 or Anaerobic Digestion Bioreactor, these terms “Anaerobic Digester” and Anaerobic Digestion Bioreactor” are used interchangeably herein. The anaerobic digester (AD) 300 may be, for example, an Up-Flow Anaerobic Sludge Blanket ((UASB) or an anaerobic membrane bioreactor. Anaerobic bacteria include, for example, protozoa, and fungi, such as, for example, Cellulolytic Bacteria, Starch-Digesting Bacteria, Proteolytic Bacteria, Methanogenic Archaea, Lactic Acid Bacteria, and Sulfate-Reducing Bacteria. The anaerobic bacteria was, for example, previously populated in the anaerobic digester reactor 300 as a starter.

In this anaerobic digestion bioreactor 300, the wet biomass is further processed, such that the biomass, having cyanobacteria cells as protein is broken into two components, Methane gas (CH4) or biogas, and Ammonium (NH4) and water, in a nutrient effluent. The methane gas (biogas) (arrow 301) is used to power the energy generator 304.

The energy generator 304 provides heat (arrow 302) to the Anaerobic Digestion Bioreactor 300, for example, to enhance process sustainability creating a circular economy system. The energy generated from the energy generator 304 is sent to the energy microgrid to power the fertilizer system 120 and may also be used to power the Photobioreactor System 110 components, such as pumps (P).

Once a processing cycle in the Anaerobic Digester bioreactor 300 is complete (the process in the anaerobic digester bioreactor 300 is a continuous process, e.g., a continuous flow process, of potentially an infinite number of cycles), the nutrient effluent is sent or pumped, via a pump (P) 305 over a line 306a to a tank 308 of the like for collecting the product, for example, an intermediate product, which is, for example a liquid solution (e.g., Algae slurry) of Ammonium (NFL) and other minerals. This Ammonium is an intermediate product, suitable as fertilizer, also known herein as Bio Liquid Fertilizer (BLF). This intermediate product may also be made into Ammonium Nitrate (NH4NO3), also suitable as a fertilizer.

Alternately the nutrient effluent, which includes Ammonium (NH4), may be pumped, via the pump (P) 305 via line 306b into an optional nitrification bioreactor 310. In the nitrification bioreactor 310, for example, there is included, typically aerobic bacteria, such as nitrification bacteria, including, for example, aerobic bacteria of the family Nitrobacteraceae and bacteria, which convert Ammonium to Nitrates, such as, for example, Nitrosomonas, Nitrosospira, Nitrosococcus, and Nitrosolobus. These bacteria convert the Ammonium (NH4), in the nutrient effluent, into Nitrates and/or Calcium/Potassium/Magnesium Nitrate, as a final product, such as fertilizer (i.e., the product sent over the line 316, via the pump (P) 315).

Alternately, the product, i.e., a nitrate -based product, may optionally be further treated (concentrated/dewatered) for packing in containers for shipping by sending it over a line 317, via the pump (P) 315, into a Reverse Osmosis Unit 320. In the RO unit 320, the product effluent (Nitride effluent) is processed, for example, by being dewatered (to concentrate the product), into a concentrated liquid fertilizer, to reach up to 5% total Nitrogen (N2)-content as a product, concentrated BLF. The product may be packaged or provided to a local or remote rotating field system hydroponic system, or any other fertigation system, such as the rotating field system 102, as shown in FIG. 1.

In the system 120, the energy used to power the components, controller 141, pumps (P) and valves (V), taps, sensors, heaters/coolers, drains, timers, and computer modules for triggering events/responding to events, is from the energy microgrid, and the controller 141 provides computer control of all of the components, pumps (P) and valves (V) of the system 120.

FIG. 3 is a diagram of the photobioreactor (PBR) 200. The PBR 200 is formed of rows 350, e.g., one row 350 shown, of columns 352. For example, one row 350 includes sixteen columns 352, but this number may vary from one to greater than sixteen. The row 350 includes a manifold 354, which holds the columns 352.

The columns 352 are, for example, each cylindrical and circular in cross section, to receive materials in water or other liquid solution, and are transparent and/or translucent to light, including sunlight (e.g., to provide energy for photosynthesis). For example, the columns 352, are approximately 2.5 meters in height, and of an inner diameter of approximately 10 or 11 cm, and made of a material such as glass or polymethyl methacrylate (PMMA). The columns 352 are typically open to the ambient environment at their upper ends 353, but may be closed by lids or other closure structures.

The columns 352 are individually connected at the bottom to one or more ports 356, which serve functions, including, for example, as a liquid inlet for adding or removing culture volume (from medium storage tank 230), and to a gas pipes for feeding an air (ambient or purified from the intake 222, including from the source 220, at a flow

adequate for culture mixing. The ports 356 may also include valves and/or pumps, taps, drains, airlifts (bubblers and/or gas injectors, for example, for CO₂ and/or other gases), to lift air and other gases into the respective column 352, for example, to agitate or stir the liquid in the column 352. The valves and/or pumps, taps, and/or drains (as well as timers and/or computer modules for triggering events/responding to events, in some cases) function together, for example, to move the various components into and other components, including cyanobacteria cultures for harvesting, out of the respective column 352. The manifold 354 valves, pumps, taps drains and/or airlifts, may be controlled fully or partially by the controller 130, 140. Each port 356 receives an internal line(s) (not shown) from the manifold 354 (extending through the manifold 354), through which gases, liquid solution and/or medium enters, and cultures with cyanobacteria (cyanobacteria (cell) culture), leaves the respective column 352.

The manifold 354 includes an inlet 357 for receiving electrical power, from the electricity microgrid, and is appropriately wired for electricity distribution throughout the row, for proper operation of the photobioreactor columns 352. The manifold 354 also includes plural inlets 358, for example, for receiving lines from the Inoculum reactors 250, lines for CO₂ 221b, ambient and/or purified/filtered air 225b, and medium (e.g., in liquid/water solution) 233b, respectively, as well as outlets 359, through which processed biomass including cyanobacteria, is sent out from the photobioreactor 200, to additional processing, or to the Harvesting Unit 270, via line 267b.

The manifold 354 is, for example, a 5-zoll pipe with low cups holding each column with a thin seal. Air (e.g., ambient air as a source of Nitrogen (N2) and Oxygen (O₂)), as well as Carbon Dioxide (CO₂) is, for example, being fed into each column 352. The manifold 354 forms fluid connections with each port 356, for example, through pipes, for example, of 1/2 zoll.

The operations of the manifold 354 and its components, including pumps, valves, taps, sensors, heaters/coolers, drains, timers, and computer modules, for triggering events/responding to events, are controlled by one of the controllers 130, 140, and are powered, for example, by the electricity microgrid.

FIG. 4 shows the photobioreactor 200 in a set up for cultivation process management. The columns 352 (of a row 350) are in communication with pump heads (H1-H3) (pumps (P)) 402 (via the manifold 354, which is not shown) that move cyanobacteria cultures (e.g., in medium and solution) from the respective column 352 into culture night keep (CNK) tanks 404, when storage at selected temperatures and atmospheric conditions is needed, for example, temperature regulation. From the columns 352, the pumps H1-H3 402, collectively pump 267a (FIG. 2A), can move selected volumes of cultures (e.g., in medium and solution) into the harvesting unit 270, while maintaining desired cyanobacteria culture amounts and at desired densities in the photobioreactor 200 (e.g., columns 352).

The harvesting unit 270, for example, receives a biomass concentration in a storage tank 410 (from the photobioreactor 200/photobioreactor column(s) 352), and filters the biomass in the filtration unit 412, for example, a centrifuge, an ultrafiltration unit, or a cross -membrane system. The time of biomass harvesting and the amount of biomass to be harvested may be determined by a machine learning (ML) model, as detailed above. The filtration unit 412 produces a product, the biomass, for example, as an algae concentration 414, and an effluent including sterile media including cyanobacteria, which is sent to a storage tank 416, over line 368x for recovery and reuse, which is then returned to the respective columns 352 (of the photobioreactor 200). over line 368b (pumped by the pump (p) 368a (FIG. 2A)) for reuse.

A cleaning solution may also be fed through the respective pump heads 402 into the columns 352, or the CNK tanks 404 whenever required. This allows for the cleaning of all surfaces (e.g., internal surfaces) of the columns 352, CNK tanks 404, and the like.

A CIP procedure is described in detail in FIG. 5, to which attention is directed. In this procedure, all piping (lines), valves, tanks, and equipment accessing or accessible by the cyanobacteria culture in solutions needs to be accessible to the CIP solution and washing water, with all washing and treatment solutions recovered, filtered, reconstituted, and re-used, as long as quality as monitored by in line salinity and turbidity measures remains acceptable. Fresh culture or medium will be provided through the same pump head, typically in only one direction.

The CIP procedure is, for example, initiated automatically either after harvesting of the whole contaminated culture, whereby the entire system of the photobioreactor columns 352 and Culture Night Keep (CNK) tanks 404 and all connecting pipes are flooded with a hypochlorite solution from the CIP tank 514, and subsequently washed with water, from the water tank 226. In case only biofilm needs to be removed, and cyanobacteria cultures (e.g., of cyanobacteria cells) can be moved to the CNK tank 404 while the photobioreactor 200 (e.g., one or more columns 352) is being treated by CIP, and the CNK tank 404 can be subject to CIP while the culture (biomass) is in the photobioreactor 200. Both the CIP solution and the washing water are beingrecovered in recovery tanks 514a (CIP regeneration) 516a (water recovery), ultra-filtered and reconstituted for re-use, to minimize water use and environmental pollution. If salinity reaches a critical threshold, chlorite is neutralized and the spent water (QC blowdown or waste water in FIG. 2A) is discarded or used for other purposes, not related to the disclosed procedure.

FIG. 5 is also a schematic representation of a PBR operational column(s) 352 and procedures with key peripherals: a column 352 of a defined volume is connected via a pump head (Hl) 502 to a Culture night keep (CNK) tank 404. Fresh starter culture and fresh growth medium can be introduced via the same head 502 (not shown), from the Inoculum reactors 250. The total culture volume can be moved from the reactor column 352 into the CNK tank 404 for temperature regulation. Whenever the whole volume is in one or the other unit, the empty unit and connected piping (lines 510 and 518a which form a cleaning in place pathway, lines 512 and 518b are the pathway of a water washing cycle) are subject to CIP (Cleaning In Place) and washed with solutions from a CIP tank 514 and a washing water tank 226. Those solutions are being regenerated from respective tanks 514a, 516a by processes such as ultrafiltration (arrows 518a, 518b), quality control (QC Blowdown) and reconstitution tothe original concentrations. CIP solution, in tank 514, and washing water solutions, in tank 226 are, for example, reconstituted and filtered by ultrafiltration and reconstituted to their original concentrations, in respective tanks 514, 226, for reuse. In the case of the water recovery tank 516a, water is periodically disposed of when qualityis deteriorating. This entire procedure will be repeated as required to avoid formation of biofilm, and accumulation of contaminations in the cultures. Every production unit will be composed of multiple independent reactors 200, whereby the CIP tank 514 and the washing water tank 226 may be used for several PBR columns 352/CNK tanks 404.

In general, all piping (to, from, and internal to, the manifold 354) and columns 352 are subject to CIP routinely and in regular intervals, with every tap, pipe, and corner, requiring regular periodic treatment before any biofilm becomes inert.

EXAMPLES

The following Examples describe various compositions, components and or procedures used in the operation of the various components disclosed for the Photobioreactor System 110 and the Fertilizer System 120, for example, as shown in FIGs. 2A, 2B and 3, as described above.

Anabaena Cultivation Procedures For The Photobioreactor 200

Axenic backup cultures

Axenic backup and starter cultures were maintained independently in three different photobioreactor 200 arrangements, e.g., independent columns, and renewed from stock cultures in Erlenmeyer flasks in the incubator shaker.

Starter cultures were maintained in single glass or Polymethylmethacrylate (PMMA) columns (column units) 352 diluted with sterile medium under sterile conditions and illuminated by Eight Emitting Diode (LED) lights at intensities of, for example, 50-120 watts/square meter, under increasing light intensity with increasing cell density. Those cultures were operated strictly separately and covered with a sterile cap to maintain maximum sterility.

Outdoor starter columns of 20% reactor volume were managed individually and were covered with anti-dust and insect netting. Direct feed lines from the starter cultures to the reactors 352 were maintained sterile, and medium supply was provided from separate piping and pumping systems.

Microscopic Supervision

A healthy culture of Anabaena, for example, was formed from Anabaenaflos aqua, Anabaena siamensis, Anabaena azollae, Anabaena variabilis, and mutant strains thereof, the Anabaena a genie of the family Nostocacae, which included predominantly of Anabaena filaments that include heterocyst cells.

It was believed that large percentages of broken filaments were an indication of a serious stress event and possibly imminent collapse of the culture, i.e., cyanobacteria culture.

It was believed that such breakage is often associated with release of organic nitrogen and organic matter into the growth medium, favoring the growth of green algae, fungi, bacteria, and predatory protozoa. If a substantial proportion of those phenomena is observed, it was believed that the culture may be collapsing.

The remediation to avoid the aforementioned collapsing included harvesting such cultures completely and feeding into anaerobic digestion, with subsequent Cleaning In Place (CIP) treatment of the reactor (reactor column 352) affected, and restarting from the high quality stock (culture) in the reactor, or in the LED columns 352.

Photobioreactor (PBR) 200 Operation

Medium including water and dissolved minerals was provided from a medium mixer, which mixes, for example, three stock solutions (A, B, C, 1 ml/1 each) directly with the desired volume of fresh, for example, reverse osmosis (RO) water, as shown below in Table 1. Table 1

Photobioreactor (PBR) Start-up

Fresh reactor columns 352 were started with an adequate volume of 20 to 30 liters of liquid, sent from the Inoculum Reactor 250, either outdoors, or indoors, including cyanobacteria cultures. The cyanobacteria cultures were, for example, grown in glass or PMMA columns 352.

For example, cyanobacteria (cell) cultures were started in a cleaned CIP treated column 352, as per FIG. 3, with adequate bubbling speed that depended on the culture height in the respective column 352. Starter culture was be added from the reactor 250 or another reactor (automatic transfer), or added from indoors through the Culture Night Keep (CNK) tank 404.

The cyanobacteria culture volume was increased by adding fresh medium slowly, for example, 10% in winter to 30% in summer of culture volume until the column (reactor) 206 is full. Automatization was available to perform this procedure remotely. Once the column (reactor) 352 was full, culture dry weight was be determined;

When dry weight reached 1.5 grams per liter in summer, or 1 - 1.2 gram per liter in winter, harvestingand dilution cycles were initiated to remove between 10% of culture volume in winter and 30 - 40% of culture volume in summer, refilling the reactors 200 (columns 352) with fresh medium.

Culture quality supervision control and monitoring Cyanobacteria culture quality, for example, was assessed visually by judging the color of the culture, which needed to be blue-green and not yellow-green. This could also be done by color sensors, which analyze colors, and include cameras, such as spectral cameras, hyperspectral cameras, or spectrum analyzers.

Excessive foam was a sign of an unhealthy culture. Excessive biofilm formation indicated starting problems with the cyanobacteria culture quality. Microscopic examination was used to reveal critical contamination or quality problems.

Cleaning In Place (CIP)

In general, all piping (to, from, and internal to, the manifold 354) and columns 352 were subject to CIP routinely and in regular intervals, with everytap, pipe and corner requiring regular periodic treatment before biofilm becomes inert.

To manage biofilm formation and biological contaminations, the procedures were employed, as disclosed above with reference to FIG. 5.

Contamination may occur when cultures are brought to high biomass concentrations, and when subject to excessive physical or chemical stress. While excessive physical and/or chemical stress was brought under control to a substantial extent, high culture density was a requirement for efficient cultivation, and thus, a balance between this and maintaining high culture integrity was defined.

Procedure for determining an amount of algae biomass for harvesting

1: Determine the amount of algae biomass (e.g., biomass) that was harvested:

Weigh the centrifuge cylinder empty before harvesting - We

Weigh centrifuge cylinder full after harvesting - Wf Difference Wf - We is the amount of algae biomass harvested M in kg or g;

Determine the volume of algae culture harvested V in liters; M/V gives wet biomass concentration in the culture in g/1; 2: Determine the dry weight of algae harvested:

Prepare an aluminum paper dish; weigh in the analytical balance.

Add 1 tablespoon of algae paste, spread equally, and weigh;

Weight of aluminum with biomass - aluminum dish empty = weight of algae biomass; Dry dish with paste in the BioloMix and weigh repeatedly until constant weight.

3. Determination of culture dry weight:

Filter 5 ml culture onto a 4.7 cm Grade C glass fiber filter weighed dry beforehand.

Dry the filter in the microwave at 30% power for 8 minutes;

Weigh the full filter, take the difference in milligrams, and divide by 5 to get the Dry weight in -milligrams per ml.

Process engineering

A 3000 liter photobioreactor facility with an associated anaerobic digester 300 was designed to supply fertilizer to a 120 square meter hydroponics unit, e.g., a rotating field system 102. No differences in the growth of lettuce and other crops fertilized with this fertilizer and synthetic control fertilizer were observed.

Photobioreactor (PBR) engineering

Biomass productivity and suitability of Anabaena cultivation outdoors under Israeli climatic conditions was tested in a 2.5 liter, 8 cm glass column approach in Sede Boker, Israel (Fig. 2A). Biomass productivity in Month 1 (August) was 0.35 - 0.4 grams/liter (g/1) per day, in Month 2 (September) 0.3 g/1 day, confirming productivity projections of around 0.3 g/1 day in the annual average under efficient light exploitation and cultivation conditions.

A scalable PBR

An airlift column photobioreactor design was chosen for cyanobacteria (CB) (e.g., cells) cultivation at scale based on commercial availability of 10 and 11 cm diameter, 2.5 m high PMMA columns 352 under attractive conditions. A reactor manifold 354, allowing to connect 16 to 19 single columns into a single PBR unit (column 352), such as that shown in FIG. 3, of 300 volume was designed allowing for simultaneous filling or harvesting ofthe reactor. Each such reactor is connected separately or in groups to a management manifold where culture can be withdrawn to harvesting, and refilled with fresh medium, activities that were controlled automatically.

The guiding principle for photobioreactor 200 engineering was the ready availability of industrially produced low cost standard parts providing a high illuminated reactor surface together with easy automatic management of reactor initiation, supervision, and CIP. The reactor columns 352 are connected to the necessary manifolds 354, operated by automatic taps, pumps, valves, and connections, to allow automatic culture transfer between tanks and reactors, to the harvesting equipment, and refilling of the columns 352 with prepared growth medium.

In case of observable culture deterioration, e.g., easily visible foam or biofilm formation or microscopically detectable culture contamination, the entire photobioreactor 200 content was harvested and transferred to Anaerobic Digestion (AD) 300, and the photobioreactor 200 content was subject to CIP (Cleaning In Place) using diluted hypochlorite solution overnight. Subsequently the reactor was washed with fresh water and re-initiated with fresh culture medium and inoculated with a partial volume of an inoculum reactor 250 of adequate quality, or from a sterile indoors starter culture. The full process from contamination detection, CIP procedure, washing and reactor column 352 re -population was a pre-programmed automatic procedure, and with the system robot capable of visually identifying critical culture quality parameters such as foam or biofilm formation, and was initiated both autonomously and automatically, thus dramatically reducing the necessary presence time of human expert system operators.

Based on a year-long operational experience, a manifold 354 was designed to function specifically by removing corners affected by biofilm formationand facilitating cleaning and maintenance, available in series at significantly lower cost. Equal resource supply into all columns 352, by varying medium supply hole diameter, and improved ease of assembly and reliability (Fig 3). Connection caps (not shown) between the manifold supply pipes for culture-medium and air-CCh, and the reactor tubes were designed and were installed into the manifold assembly 354, and provided more precise flows of liquid and air throughout the whole photobioreactor system 110, for enhanced reliability and reproducibility of the cultivation process.

Contamination Control and Temperature Control

The system 110 may also undergo stress factors, which may affect culture stability, which may lead to complete and rapid culture collapse of whole reactor units. Specific modifications to the facility engineering, process flows, and management were introduced and tested successfully to overcome most of those problems. These three factors, individually, or in combination may cause lysis of the Anabaena filaments under lysis and release of cellular content, permitting growth of symbiotic and parasiticmicrobial contaminants or complete lysis of the whole culture content. The following stress factors were observed: a) Chemical stress: Chemical stress due to occasionally inadequate water quality, culture stability apparently caused by fluctuating water quality. This problem was overcome by introducing a regular manifold rinsing procedure via pumping a partial culture volume out into a storage tank (e.g., CNK tank 404) and returning the mixed volume back into the reactor 352. An alternative procedure was successfully applied as pumping defined volumes of fresh medium through the manifold 354 into the columns 352, resulting in maintaining a steady culture concentration while refreshing mineral content and compensating for evaporative losses. Procedures for both cases were programmed into the process management software (discussed below), which can be remotely activated and controlled, and adjusted, as required or necessary. b) Physical stress: Physical stress due to excess radiation and temperature stress: Excess radiation can be detrimental to algae cultures. However, in the reactor 200, adequate dilution speeds and critical culture densities were identified. Temperature was a more critical factor to manage and control. A procedure called a ‘night keep’ has therefore been developed, consisting of removing the whole reactor content intoan insulated tank 404 for temporary storage until adequate temperature is restored or external conditions become favorable again. During cool winter days this becomes critical as cyanobacteria of the cyanobacteria cultures are subject to photo-oxidative stress when they are exposed to light when at iboptimal temperature. Therefore, the warm afternoon cultures are pumped into the insulated night keep tank 404 where temperatures of around 25 degrees can be maintained overnight, so that in the morning after sunrise, temperate cultures were returned into the columns 352 for immediate onset of maximal photosynthetic efficiency. During days of excessive cold, the cultures in the night keep tanks 404 were warmed by heat exchange using warm water, created by solar water heaters. The same procedure was applied to cool down the cultures in summer, when cultures approaching the critical limit of 40 degrees C were pumped into the insulated tank 404, which were cooled by a mantle with circulating cooler water until cooled sufficiently (to about 35 degrees C) to be returned into the columns 352 . This procedure can be repeated as required to prevent cultures from exceeding 40 degrees centigrade. The necessary cooling water was generated by evaporative effect while circulating over cooling matrices as commonly used in greenhouses, but avoiding excessive evaporation and recovering the cooled water. A further note of importance is temperature stability of medium added, Anabaena can react sensitively to dilution with growth medium provided at a temperature strongly differing from the culture temperature, and growth medium must therefore be temperate to the range of the culture temperature before addition. A further problem observed was precipitation of medium salts in stock solutions under storage. Thus, all of the stock solutions were mixed routinely before supply to the cultures. c) Biological stress: Biological stress occurred due to competition, predation, and specific microbial pathogens Optimization of cultivation procedures:

It was desirable to prevent and otherwise inhibit any biofilm formation in the columns 352. Biofilms can rapidly build up in the columns 352. The biofilms are, for example, composed of fungal contaminants associated with Anabaena, bacteria, protozoa, and filamentous green algae that affect productivity negatively and act as breeding ground for parasitic fungi and protozoa. One solution involved frequent CIP and restarting the affected columns 352 with clean inoculum from other columns 352, all possible by means of automated pre-programmed operation using a CIP solution of 5000 ppm hypochlorite stored and reconstituted in a CIP storage tank 516, and washing with water from a washing water tank 226. Induction of the CIP and washing procedure was being programmed into the function of the system’s control system that was visually supervising the status of the cyanobacteria cultures in all of the columns 352 and transparencyof the columns 352 while harvesting.

As disclosed above, CIP solution and washing water solutions were reconstituted and filtered by ultrafiltration to their original concentrations for reuse, with periodic disposal thereof, when quality had deteriorated beyond acceptable limits.

Contamination was a potential problem when cyanobacteria cultures were brought to high biomass concentrations, and when subject to excessive physical or chemical stress. Accordingly, it is believed that high quality culture density is a requirement for efficient cyanobacteria cultivation.

Anaerobic Digestion process for fertilizer production

Biomass from the Photobioreactor system 110 was periodically harvested, by the Harvesting Unit (HU) 270, either using a flow through centrifuge or a Vibro ultrafiltration unit. The harvested algal biomass slurry (10 - 20 liters at about 20 g/1 biomass concentration) is fed into two units of two 120 liter UASB anaerobic digesters 300. The anaerobic digester 300 was, for example, operated as an Up-Flow Anaerobic Sludge Blanket (USAB) Reactor, with biomass sludge fed into the bottom of the reactor 300, and excess liquid drained from the top directly feeding a 120 square meter hydroponics unit, such as the rotating field system 102.

The anaerobic digestion bioreactor 300 effluent released from the top during transfer of algae sludge into the AD is directed into a hydroponics unit of 120 square meters, to test the effluent’s suitability as liquid fertilizer for hydroponics. The crops cultivated with this effluent were observed over a period of 12 months, included lettuce, red leaf lettuce, and basil, which performed equally well in the effluent, when compared to the same size sample, using the product from the Fertilizer System 120.

Specifically, in the rotating field system 102, no microbial contamination of the hydroponics water was observed, and the crop plants developed equally well as in the control plot, and did not show any signs of additional stress.

Ultrafiltration, concentration, and packaging

For concentration and commercial marketing, the nutrient effluent from the anerobic digestion bioreactor 300 was subjected to ultrafiltration, to remove particles, and then subjected to nitrification in the Nitrification Bioreactor 310, and then concentrated by three cycles of reverse osmosis (RO), in a Reverse Osmosis unit 320, to achieve a total Nitrogen concentration of approximately 5%. For example, a content of Ammonium (NH4) Nitrogen (N2) in the Anaerobic Digestion Bioreactor was 2200 parts per million (ppm) measured using Nessler’s reagent, in the average of three different sampling dates from two different AD reactors 300. For example, one cycle of threefold concentration was achieved to reach 0.6% of Nitrogen. Three cycles of concentration by RO created the required nitrogen concentration for successful commercialization. With 5% of ammonia N, the product was superior to any current organic nitrogen fertilizer solution, based on higher Nitrogen concentration, with higher purity and higher reproducibility, and free of any pathogens.

Product- Liquid Nitrogen-rich organic fertilizer based on renewable Nitrogen fixed by cyanobacteria

Composition of the Fertilizer Product

The cyanobacteria (CB) biomass (e.g., as wet biomass) from the Photobioreactor System 110 was transferred into the Fertilizer System 120 at 15 to 20 grams/liter into an UASB anaerobic digester (Anaerobic Digestion Bioreactor 300), whereby all of the biomass was converted into Ammonium (NH4) and biogas (CH4 and CO₂) and most Nitrogen was accumulated in the liquid phase, predominantly as Ammonium dissolved in the medium together with PO4, heavy metals, and the essential oligo-elements Ca, Mg, and S (Table 2.1). Nutrient effluent was released from the top volume of the Anaerobic Digestion Bioreactor 300 each time a volume of harvested CB slurry was inserted at the bottom of the UASB reactor (Anaerobic Digestion Bioreactor 300). That nutrient effluent was analyzed and found to be a highly suitable organic liquid fertilizer.

Table 2.1: ICP results samples, filtered, in ppm

Table 2.1

Table 2.1 provides concentration ranges of Nitrogen, and key element concentration in the anaerobic reactor effluent. Significant amounts of heavy metal and oligo-element ions are missing from the mass balance, apparently precipitating under the conditions of the anaerobic reactor. Such precipitate was recovered, dissolved, and used as fertilizer supplements as required.

Upon further anaerobic digestion (fermentation) of the biomass in the anaerobic digestion bioreactor 300, Nitrogen concentrations were in the Nitrification Bioreactor 310, up to 0.25 to 0.3% (see Table 2.2) to present an in line organic liquid fertilizer.

Due to the highly controlled automated production process, a well-defined fertilizer composition was obtained, that was field tested as the sole fertilizer supply for a hydroponics unit, which, for example, produced various lettuce species and different herbs.

A number of elements, specifically after clarification by Nitrification and, filtration or centrifugation, were underrepresented and were found in the precipitate as struvite, or insoluble heavy-metal compounds. All precipitated and filtered residues were recovered, dissolved in organic acids (e.g., citric acid) and added to the liquid fertilizer, found as a separate organic oligo-element supplement for agriculture. Table 2.2: Nitrogen determination of AD effluent

Table 2.2

Effluent (nutrient effluent, also referred to here as AD effluent) from the anerobic digester 300 was determined using the Nessler’s Reagent (K^HgE) testing method, after different pretreatments: In the average of three independent samples, an average Nitrogen (N) content of 0.22% measured as ammonia was determined. Those samples were concentrated three-fold by reverse osmosis and analyzed at the Neve Ya’ar experimental station of the Ministry of Agriculture of Israel.

Table 2.3

Table 2.3 provides an analysis of 3-fold concentrated AD effluent. The sample contained 0.6% N, 70% of which as free ammonia, with 30% organic amines and without nitrate. Phosphate content as well as Ca and Mg were adequate for a liquid fertilizer. Considering the overall material flow through the system, harvesting 750 gram biomass per day (the expected daily output from the system (Alpha Prototype from Growponics Ltd. of Kyriat Bialik Israel)), containing about 60 grams of Nitrogen, yielded about 30 liters per day of liquid fertilizer at 2 grams of Nitrogen per liter.

In situ test of fertilizer quality

Field testing of the full value chain including continuous fertilizer production and application to actual greenhouse hydroponics cultivation as compared to conventional fertilizer application was performed. No difference in growth and production of lettuce and other crops evaluated as observed during the whole test phase of 18 months, between the hydroponics unit fed with commercial synthetic fertilizer, as compared to a unit fed directly with the AD effluent derived from cyanobacteria (CB) biomass digestion.

All of the sensors and actuators (pumps, valves, taps, sensors) in this process were controlled with a system 600 (FIG. 6), which controlled the processes performed by the Photobioreactor System 110, and, for example, is operated by a LabVIEW application that is now in its first and most basic version REV 4.0. This system 600, for example, permitted autonomous automatic process operation for most of the duration, with the majority of possible disturbances handled, and also allowed for remote control of all processes.

An integrated robot visually supervised the status of the photobioreactor 200. i.e., the photobioreactor columns 352, supervising biomass and fertilizer development and biofilm formation towards initiation of critical maintenance steps such as initiating complete harvesting, CIP, and restarting of photobioreactor columns 352, and performing other process control functions.

Photobioreactor (PBR) System 110 Artificial Intelligence (Al) Control Software

The Photobioreactor 200 included an advanced Artificial intelligence (Al) control and management software, shown in FIGs. 6-10, to which attention is directed. The software was, for example, written in LabVIEW. It utilized object-oriented programming focused on various types of Input-Output (IO) modules. User-programmable procedures interacted with the modules to provide high flexibility and functionality to the photobioreactor 200 operation. Communication with a Remote Input Output (RIO) was conducted over a serial Universal Asynchronous Receiver-Transmitter (UART) protocol in a dedicated RIO Engine sub- VI (virtual instrument). The software functioned as standalone software or as a nested application within comprehensive greenhouse management software.

FIG. 6 is a screen shot of an overview screen 602, which displays all active modules in units selected by the user. From this screen, modules can be added, deleted, or edited using a dedicated faceplate, as seen in FIG. 7. Modules are shown, for example, associated with the medium tank 230, the row 350 of photobioreactor columns 352 of the manifold 354, and for various processes of the individual photobioreactor columns 352. Other stand-alone modules operate to perform, for example, harvesting, pumping, and cleaning in place.

Each Input Output (IO) unit was represented by a module, either analog or digital, as shown for example, in FIG. 8. The modules were, for example, input modules, such as pH or temperature sensors, color sensors, which analyze colors, such as cameras, including spectral cameras, hyperspectral cameras, or spectrum analyzers, or output modules like valves and pumps (pump actuators). There were also function modules for advanced calculations and functionalities. Modules were uniquely routed to a Remote IO (RIO) address and port. They maintained their process parameters, which were saved in the module list files. The modules' process parameters were recorded in data files.

The software (algorithm), for example, supported up to 100 RIO units, each with its own modules. FIG. 8 is a diagram of the modules' hierarchy.

Procedures and Al

Procedures included action items initiated upon certain triggers and as calculated from previous procedures ( i.e., machine learning (ML)). When the required number of triggers was met, either at the beginning or end of the procedure, a list of action items was engaged for the execution of process actuators. The Procedure Engine 710 (FIG. 10) was a sub VI responsible for collecting triggersand initiating action items. FIG. 9 illustrates an example of one of the triggering process procedures. Each procedure is saved as an XML file and can be loadedand edited by the user.

Timing Handling

The Time Engine (timer) 712 (FIG. 10) handled time -related triggers and constantly checked whether they are due.

Triggers and Cascades

Triggers can be time-related (defined by the hour or daytime indicator) or activated by a sensor or function modules. They can also activate a specific time after a procedure has started or ended.

Every module and procedure had a cascade list that stored all potential triggers that might initiate upon a certain cue. When a module detects the appropriate cue, such as rising above a specific value, it sent a command to the procedure engine 710 to engage the correct trigger, as shown, for example, in FIG. 9.

Relationships scheme

FIG. 10 shows the main PBR VI 702 (a screen oriented text editor) hosts the Graphical User Interface (GUI) and managed user commands. Modules, procedures, and settings, for example, an event handler 704, a command handler 705, a Remote Input and Output (RIO) server 706., and a drag and drop functionality 707 for the GUI, can be viewed and edited from this VI. Two process engines ran in parallel: the procedure engine 710, which handled procedure triggering and action item operations, and the time engine 712, which oversees all time-related processes and triggers. The Remote Input Output (RIO) Engine 720 managed communication with the RIO.

The wide variety of modules can represent any process sensor or actuator that can connect to the RIO Engine 720, and the procedures can adapt to any operation standard and scheme, for example, data analysis 722, Al analysis an event handler 723, and a command handler 724. A read/write module 726 is, for example, internal or external to the RIO Engine 720, to engage a correct trigger. The software (algorithm) already logged process parameters and supported user selection, alarms, and other essential features for GoodManufacturing Process (GMP) operation.

SYSTEM OPERATION

Attention is now directed to FIGs. 11A and 11B, collectively, FIG. 11, and FIG. 12. These drawing figures are flow diagrams of example operations performed by the Photobioreactor System 110 (FIG. 11) and the Fertilizer System 120 (FIG. 12). Reference is also made in the descriptions below to components in FIGs. 2A, 2B and 3. Both systems 110, 120 and their respective pumps 160, valves 161, sensors 162, components 163, such as taps, drains, heaters, coolers, timers, triggers, and others, and manifold components 164 used for their operation are controlled by the respective controller 130, 140, 141.

Turning to FIGs. 11A and 1 IB, there is shown an example operation for the Photobioreactor System 110. The process begins at a START 1102, where the Photobioreactor System 110 begins to prepare the photobioreactor 200 (representative of the bioreactor columns 352), for example, to grow cyanobacteria, for example, which while growing performs Nitrogen fixation in its cells.

The process moves to block 1104, where an inoculum charge 252, for example, a cyanobacteria culture, is injected, for example, as a starter culture, into the photobioreactor 200, from the inoculum reactor 250. The process moves to block 1106, where a determined amount of new medium, from the medium tank 230 is populated, e.g., by pumping, into the photobioreactor 200. The medium includes, for example, nutrients, such as macro and micro nutrients, in a liquid, e.g., water, solution. Carbon Dioxide (CO₂) is also brought into the photobioreactor 200, at block 1108, for example, at pressures sufficient to bubble or airlift the liquid solution, so as to agitate or stir it in the photobioreactor 200. Ambient air, as a source of Nitrogen gas (N2) and Oxygen gas (O₂), is brought into the photobioreactor, at block 1110. The order of blocks 1106, 1108, 1110 may be as shown or different, and may occur in any order simultaneously or contemporaneously.

The photobioreactor 200 is exposed to light, such as sunlight 202, at block 1112. The now populated photobioreactor 200, is such that the exposure to sunlight 202, at block 1112, provides energy to activate and drive photosynthesis in the cells of the cyanobacteria culture. The photosynthesis results in carbon capture and sugars being formed, which provide the energy for cyanobacteria cells in the culture to perform nitrogen fixation, resulting in Ammonium in the cyanobacteria cells as the cyanobacteria culture grows.

The process moves to block 1114, where it is determined whether there is a trigger to harvest biomass from the cyanobacteria culture and nutrients in the liquid solution in the photobioreactor 200. Triggers may be based on culture density detected inside the photobioreactor 200, by sensors for color, temperature (internal/external), pressure, volume, pH, internal/external humidity, and others. For example, a threshold density for the cyanobacteria culture may be about 1 to 2 grams/liter, this range being a trigger to harvest the culture as biomass, removing a determined mount of biomass from the photobioreactor. The amount of biomass for removal, and in some cases, the exact time for removal, may be determined by the Machine Learning Model or a computer program, or in some cases, manually. The model and the program may also calculate a sufficient amount of culture to remain in the photobioreactor 200, so that the culture is healthy and will resume proper growth post harvesting.

Absent a trigger, at block 1114, the process returns to block 1112, from where it resumes. When there is a trigger, the process moves to block 1116, where the determined amount of biomass to be harvested by the Harvesting Unit (HU) 270, in the determined amount, for example, at the determined time. The process moves, for example, simultaneously or contemporaneously, to blocks 1118a and 1118b.

At block 1118b, a calculation is made, for example, by the controller 140, of the amount of medium lost with the harvest of the biomass from the photobioreactor 200. At block 1118b, the machine learning model is updated with parameters associated with the biomass harvest, such as amount, time, density, temperature (internal/external), internal/external humidity, pressure, pH, and others.

From block 1118b, the process moves to block 1120, where the harvested biomass is filtered, for example, to reduce its concentration, in some cases dewater the biomass, and filter out algae and cyanobacteria in liquid solution with medium, to be used in replenishing the photobioreactor 200, for example, as shown and described for FIG. 4. This filtered cyanobacteria in liquid solution with medium is known as regenerated medium, as per block 1126.

Returning to block 1120, the process moves to block 1124, where the filtered biomass may be dried, for example, in dryers, so as to be dry biomass, or not dried, so as to be wet biomass. These biomass products may be moved into the anaerobic digester reactor 300 of the Fertilizer System 120, for processing into, for example, Ammonium and fertilizer.

Returning to block 1120, the process moves to block 1126, where it is determined whether there is regenerated medium from the filtration in the Harvesting Unit 270. If yes, the process moves to block 1128, where the amount of regenerated medium is determined to replace the determined amount of lost medium. The process then moves to block 1130, where it is determined whether an amount of new medium is needed in addition to the regenerated medium to replace the amount of lost medium, as the amount of regenerated medium may be insufficient. If no at block 1130, new medium is not needed for the photobioreactor 200, and the determined amount of regenerated medium is added to the photobioreactor 200, at block 1132, from where the process resumes from block 1108.

If yes, at block 1130, the process moves to block 1106, where a determined amount of new medium is added to the photobioreactor 200, and then to block 1132, where a determined amount of regenerated medium is added to the photo bioreactor 200. The process then moves to block 1108, from where it resumes.

Returning to block 1126, should there not be any regenerated medium, the process moves to block 1134, where the amount of new medium to replace the amount of lost medium is determined. Upon this determination, the process moves to block 1106, where the determined amount of new medium is returned to the photobioreactor, and then to block 1108, from where the process resumes.

Should additional cyanobacteria culture be at the time of the performance of blocks 1130 and 1134, the inoculum charge of block 1104 may occur.

Although one cycle of the process performed by the Photobioreactor System 110 has been described, the process is continuous and potentially infinite. Accordingly, multiple cycles may be performed as desired.

Turning to FIG. 12, an example operation for the Fertilizer System 120 is shown. The process begins at a START 1202, where the Fertilizer System 120 and in particular, the Anaerobic Digestion Bioreactor 300 is made ready for the harvested biomass or treated harvested biomass, referred to here as the “biomass”, and typically wet biomass, from the Photobioreactor System 110. The process moves to block 1204, where the wet biomass is provided to the Anaerobic Digestion Bioreactor 300, for example, by pumping or other form of transport and/or placement, so that the biomass is fed into the Anaerobic Digestion Bioreactor 300. Anaerobic digestion of the biomass in the anaerobic digestion bioreactor 300 occurs at block 1206. In the Anaerobic Digestion Bioreactor 300, the biomass (e.g., wet biomass) is digested by anaerobic bacteria, resulting in Ammonium (NH4) being released from the cyanobacteria cells of the biomass, as well as the emission of methane gas (CH4) from the cyanobacteria. The Ammonium, having been subjected to the anaerobic bacteria, is in a nutrient effluent in the anaerobic digestion bioreactor 300, ready to be released from the anaerobic digester reactor 300.

The process moves to block 1208, where a trigger or event occurs indicating the need to remove a portion or all of the nutrient effluent, as determined by a computer program, machine learning model, manually, or a combination thereof. The trigger or event may be a specific time (a timer activating a pump to remove nutrient solution), based on volume in the Anaerobic Digestion Bioreactor 300 (from a sensor inside the Anaerobic Digestion Bioreactor 300, which causes the controller to activate a pump to remove nutrient effluent). Conditions in the Anaerobic Digestion Bioreactor 300, such as temperature, pressure, and the like.

Should there be no trigger or event, the process moves to block 1206, from where it resumes. If there is a trigger or event, the process moves to block 1210.

At block 1210, the removal process for the nutrient effluent occurs. The nutrient effluent may be removed to be used as an Ammonium solution, for example, suitable as fertilizer, at block 1214a, or sent to a Nitrification bioreactor 310, for nitrification of the Ammonium, at block 1214b. The processes of blocks 1214a and 1214b may be performed individually, or together, simultaneously, contemporaneously, or at different times, depending on the need of the system 120 user.

Optionally, from block 1210, the process may move to block 1212. At block 1212 an amount of biomass, to replace and replenish the removed nutrient effluent from the Anaerobic Digestion Bioreactor 300 is determined. The process moves to block 1204 where the determined amount of biomass is provided to the Anaerobic Digestion Bioreactor 300, and from block 1204, the process resumes.

Returning to block 1214b, and from block 1214b, the nutrient effluent is sent to the Nitrification Bioreactor 310, for example, via a pump 315. In the Nitrification Bioreactor 310, the Ammonium in the nutrient effluent is subjected to Aerobic bacteria and carbonates, such as Calcium Carbonate, Magnesium Carbonate, Potassium Carbonate, and other to create Nitrate salts, such as Calcium Nitrate, Magnesium Nitrate, Potassium Nitrate, and other Nitrate salts, for example, in an effluent. It is then determined whether or not, at block 1220, to treat the salt containing effluent, with processes such as Reverse Osmosis. If not, at block 1222, the product is suitable as fertilizer. If yes, at block 1224, the Reverse Osmosis treated salt containing effluent is also suitable as fertilizer, such as a BLF product, for example, concentrated BLF.

From blocks 1222 and 1224, the process ends, but may be repeated for as long, e.g., as many cycles, as desired, as the process is continuous, and potentially infinite.

Alternative embodiments of the disclosed subject matter include proteins produced with nitrogensourced from ambient air. The resultant product is a “concentrate” of protein powder that was produced without chemical fertilizers. The disclosed protein concentrates are produced using either chemical Nitrogen (they cannot be certified organic) or using organic fertilizer for manure or organic waste.

The disclosed subject matter is directed to a method for producing nitrogen-based fertilizers. The method comprises: obtaining hydrogen from water; sourcing nitrogen from ambient air as ambient nitrogen; and, feeding the water and ambient nitrogen into a photobioreactor, the photobioreactor populated with an effective amount of one or more strains of: bacteria, algae, and/or microorganisms, to maintain stability in the photobioreactor, to facilitate nitrogen fixation and photosynthesis, including carbon capture, to grow biomass, for the production ofBiofertilizer and/or protein-rich food.

Optionally, the method is such that the nitrogen fixation and the photosynthesis occur contemporaneously.

Optionally, the method is such that the bacteria includes cyanobacteria.

Optionally, the method is performed without feeding photobioreactor a sugar or any other chemical energy stream.

Optionally, the photobioreactor facilitates digestion of the resulting biomass by anaerobic digestion, and separation and treatment the Anaerobic effluent from the photobioreactor includes bio-fertilizer. The disclosed subject matter is directed to a method for producing a fertilizer product. The method comprises: in a first vessel, providing a liquid solution comprising: water as a hydrogen source; a bacteria culture, the bacteria culture in an amount sufficient such that when activated, performs Nitrogen fixation and photosynthesis; nutrients for the bacteria culture; air as a Nitrogen source for Nitrogen fixation by the bacteria culture, and an oxygen source; and, Carbon Dioxide (CO₂) as a gas. The liquid solution in the first vessel is exposed to light, including sunlight, to activate the bacteria culture and grow the bacteria in the bacteria culture. At least a portion of the bacteria culture is removed (e.g., harvested) from the first vessel as biomass (for placement into a second vessel). In a second vessel, the bacteria culture of the biomass is anaerobically digested by reacting the biomass with an amount of anaerobic bacteria sufficient to produce Ammonium (NH4) in an effluent.

Optionally, the method is such that it additionally comprises: converting the Ammonium of the effluent, in a third vessel, to a Nitrate by reacting the Ammonium with a Carbonate.

Optionally, the method is such that the first vessel includes a photobioreactor.

Optionally, the method is such that the photobioreactor includes one or more columns.

Optionally, the method is such that the bacteria culture includes a cyanobacteria culture.

Optionally, the method is such that the cyanobacteria culture includes cyanobacteria selected from the group consisting of: Anabaena flos aqua, Anabaena siamensis, Anabaena azollae, Anabaena variabilis, and mutant strains thereof.

Optionally, the method is such that the cyanobacteria culture includes cyanobacteria of the family Nostocacae.

Optionally, the method is such that the air includes ambient air.

Optionally, the method is such that the nutrients include micro nutrients.

Optionally, the method is such that the micronutrients include one or more of: Boron, Copper, Sodium, and Phosphorous.

Optionally, the method is such that the nutrients include macronutrients.

Optionally, the method is such that the macronutrients include one or more of: Magnesium, Potassium, and Calcium.

Optionally, the method is such that the providing the Carbon Dioxide as a gas is in a gas stream at pressures sufficient to cause airlift in the liquid solution in the first vessel.

Optionally, the method is such that the removing at least a portion of the bacteria culture as biomass is performed when the density of the bacteria culture reaches a predetermined value.

Optionally, the method is such that the predetermined value is at least between 1 to 2 grams per liter.

Optionally, the method is such that the removing at least a portion of the bacteria culture as biomass, includes determining the amount of the bacteria culture to be removed by a Machine Learning model.

Optionally, the method is such that the removing at least a portion of the bacteria culture as biomass, includes determining the amount of the bacteria culture to be removed by a computer algorithm.

Optionally, the method is such that the second vessel includes an anaerobic digester.

Optionally, the method is such that the anaerobic bacteria for anaerobically digesting the biomass includes one or more of: protozoa, fungi, Cellulolytic Bacteria, Starch-Digesting Bacteria, Proteolytic Bacteria, Methanogenic Archaea, Lactic Acid Bacteria, and Sulfate - Reducing Bacteria.

Optionally, the method is such that the bacteria culture is provided to the liquid solution as a starter culture in an inoculum charge.

Optionally, the method is such that third vessel includes a Nitrification bioreactor.

Optionally, the method is such that the converting the Ammonium of the effluent to a Nitrate additionally comprises reacting the Ammonium effluent with a sufficient amount of aerobic bacteria.

Optionally, the method is such that the aerobic bacteria is selected from the group consisting of: Nitrosomonas, Nitrosospira, Nitrosococcus, and Nitrosolobus.

Optionally, the method is such that the aerobic bacteria include bacteria is of the family Nitrobacteraceae.

Optionally, the method is such that the Carbonate includes one or more of: Calcium Carbonate, Potassium Carbonate, and Magnesium Carbonate.

Optionally, the method is such that the Nitrate includes Calcium Nitrate, Potassium Nitrate, and/or Magnesium Nitrate.

The disclosed subject matter is directed to a system for producing a fertilizer product. The system comprises: a photobioreactor system and a fertilizer system. The photobioreactor system comprises: a photobioreactor for growing a cyanobacteria culture using ambient nitrogen; and, a harvesting unit in communication with the photobioreactor for harvesting the cyanobacteria when grown as biomass. The fertilizer system comprises: an anaerobic digestion bioreactor for converting harvested biomass to Ammonium (NH4) in an effluent; and, a Nitrification bioreactor in communication with the anaerobic digester for converting the Ammonium in the effluent to Ammonium Nitrate.

Optionally, the system is such that it additionally comprises: a controller in communication with sensors in the photobioreactor, the controller for 1) determining the density of the cyanobacteria culture in the photobioreactor, and 2) based on the density determination, determining whether to harvest at least a portion of the cyanobacteria culture.

Optionally, the system is such that the controller applies at last one of a Machine Learning model or a computer algorithm, to determine the time to harvest the at last a portion of the cyanobacteria culture and the amount of the at least a portion of the cyanobacteria culture to be harvested.

Optionally, the system is such that the harvesting unit includes a centrifuge and/or a membrane filtration system.

Optionally, the system is such that the photobioreactor includes one or more columns of a material translucent or transparent to sunlight.

The disclosed subject matter is directed to a method for producing a fertilizer product. The method comprises: providing a liquid solution comprising: water as a hydrogen source; a cyanobacteria culture, the cyanobacteria culture in an amount sufficient such that when activated, performs Nitrogen fixation and photosynthesis; nutrients for the bacteria culture; ambient air as a Nitrogen source for Nitrogen fixation by the bacteria culture, and an oxygen source; and, Carbon Dioxide (CO₂) as a gas; and, exposing the liquid solution in the first vessel to light, including sunlight, to activate the cyanobacteria culture and grow the bacteria in the bacteria culture.

In other aspects of the disclosed subject matter, the process can be adjusted to achieve certifiable organic status, and the process is designed to be fully greenhouse gasses (GHG) neutral, and waste and emission free.

The implementation of certain methods and/or systems of the disclosure can involve performing or completing selected tasks manually, automatically, or a combination thereof. Moreover, according to actualinstrumentation and equipment of examples of the method and/or system of the disclosure, several selectedtasks could be implemented by hardware, by software or by firmware or by a combination thereof using an operating system or a cloud-based platform.

For example, hardware for performing selected tasks according to examples of the disclosure could be implemented as a chip or a circuit. As software, selected tasks according to examples of the disclosure couldbe implemented as a plurality of software instructions being executed by a computer using any suitable operating system. In an exemplary example of the disclosure, one or more tasks according to exemplary examples of method and/or system as described herein are performed by a data processor, such as a computing platform for executing a plurality of instructions. Optionally, the data processor includes a volatile memory for storing instructions and/or data and/or a non-volatile storage, for example, non-transitory storage media such as a magnetic hard-disk and/or removable media, for storing instructions and/or data. Optionally, a network connection is provided as well. A display and/or a user input device suchas a keyboard or mouse are optionally provided as well.

For example, any combination of one or more non-transitory computer readable (storage) medium(s) may be utilized in accordance with the above-listed examples of the present disclosure. The non-transitory computer readable (storage) medium may be a computer readable signal medium or a computer readable storage medium. A computer readable storage medium may be, for example, but not limited to, an electronic, magnetic, optical, electromagnetic, infrared, or semiconductor system, apparatus, or device, or any suitable combination of the foregoing. More specific examples (a non-exhaustive list) of the computer readable storage medium would include the following: an electrical connection having one or more wires, a portable computer diskette, a hard disk, a random access memory (RAM), a readonly memory (ROM), an erasable programmable read-only memory (EPROM or Flash memory), an optical fiber, a portable compact disc read-only memory (CD-ROM), an optical storage device, a magnetic storage device, or any suitable combination of the foregoing. In the context of this document, a computer readable storage mediummay be any tangible medium that can contain, or store, a program for use by or in connection with an instruction execution system, apparatus, or device.

A computer readable signal medium may include a propagated data signal with computer readable programcode embodied therein, for example, in baseband or as part of a carrier wave. Such a propagated signal maytake any of a variety of forms, including, but not limited to, electro-magnetic, optical, or any suitable combination thereof. A computer readable signal medium may be any computer readable medium that is not a computer readable storage medium and that can communicate, propagate, or transport a program for use by or in connection with an instruction execution system, apparatus, or device.

As will be understood with reference to the paragraphs and the referenced drawings, provided above, various examples of computer-implemented methods are provided herein, some of which can be performedby various examples of apparatuses and systems described herein and some of which can be performed according to instructions stored in non-transitory computer- readable storage media described herein. Still, some examples of computer-implemented methods provided herein can be performed by other apparatuses or systems and can be performed according to instructions stored in computer-readable storage media otherthan that described herein, as will become apparent to those having skill in the art with reference to the examples described herein. Any reference to systems and computer-readable storage media with respect to the following computer-implemented methods is provided for explanatory purposes and is not intended to limit any of such systems and any of such non-transitory computer-readable storage media with regard to examples of computer-implemented methods described above. Likewise, any reference to the following computer-implemented methods with respect to systems and computer-readable storage media is provided for explanatory purposes and is not intended to limit any of such computer-implemented methods disclosed herein.

The flowcharts and block diagrams in the Figures illustrate the architecture, functionality, and operation ofpossible implementations of systems, methods, and computer program products according to various examples of the present disclosure. In this regard, each block in the flowchart or block diagrams may represent a module, segment, or portion of code, which comprises one or more executable instructions for implementing the specified logical function(s). It should also be noted that, in some alternative implementations, the functions noted in the block may occur out of the order noted in the figures. For example, two blocks shown in succession may, in fact, be executed substantially concurrently, or the blocksmay sometimes be executed in the reverse order, block diagrams and/or flowchart illustration, and combinations of blocks in the block diagrams and/or flowchart illustration, can be implemented by special purpose hardware -based systems that perform the specified functions or acts, or combinations of special purpose hardware and computer instructions. The descriptions of the various examples of the present disclosure have been presented for purposes of illustration but are not intended to be exhaustive or limitedto the examples disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described examples.

As used herein, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise.

It is appreciated that certain features of the disclosure, which are, for clarity, described in the context of separate examples, may also be provided in combination in a single example. Conversely, various featuresof the disclosure, which are, for brevity, described in the context of a single example, may also be providedseparately or in any suitable sub-combination or as suitable in any other described example of the disclosure. Certain features described in the context of various examples are not to be considered essential features of those examples unless the example is inoperative without those elements.

The above-described processes including portions thereof can be performed by software, hardware, and combinations thereof. These processes and portions thereof can be performed by computers, computer- type devices, workstations, cloud-based platforms, processors, micro processors, other electronic searching tools and memory and other non-transitory storage-type devices associated therewith. The processes and portions thereof can also be embodied in programmable non-transitory storage media, for example, compact discs (CDs) or other discs including magnetic, optical, etc., readable by a machine or the like, or other computer usable storage media, including magnetic, optical, or semiconductor storage, or other source of electronic signals. The processes (methods) and systems, including components thereof, herein have been described with exemplary reference to specific hardware and software.

The processes (methods) have been described as exemplary, whereby specific steps and their order can beomitted and/or changed by persons of ordinary skill in the art to reduce these examples to practice without undue experimentation. The processes (methods) and systems have been described in a manner sufficient to enable persons of ordinary skill in the art to readily adapt other hardware and software as may be neededto reduce any of the examples to practice without undue experimentation and using conventional techniques.

Descriptions of examples of the disclosure in the present application are provided by way of example and are not intended to limit the scope of the disclosure. The described examples comprise different features, not all of which are required in all examples of the disclosure. Some examples utilize only some of the features or possible combinations of the features. Variations of examples of the disclosure that are described, and examples of the disclosure comprising different combinations of features noted in the described examples, will occur to persons of the art. The scope of the disclosure is limited only by the claims.

It will thus be appreciated that the examples described above do not limit the disclosed subject matter to what has been particularly shown and described hereinabove. Rather, the scope of the present disclosure includes both combinations and sub-combinations of the various features described hereinabove, as well asvariations and modifications thereof which would occur to persons skilled in the art upon reading the foregoing description and which are not disclosed in the prior art. Documents incorporated by reference in the present patent application are to be considered an integral part of the application except that to the extentany terms are defined in these incorporated documents in a manner that conflicts with the definitions madeexplicitly or implicitly in the present specification, only the definitions in the present specification should be considered.

Claims

1. A method for producing nitrogen-based fertilizers comprising: obtaining hydrogen from water; sourcing nitrogen from ambient air as ambient nitrogen; and feeding the water and ambient nitrogen into a bioreactor, the bioreactor populated with an effective amount of one or more strains of: bacteria, algae, and/or microorganisms, to maintain stability in the bioreactor, to facilitate nitrogen fixation and photosynthesis, including carbon capture, to grow biomass, for the production of Biofertilizer and/or protein-rich food.

2. The method of claim 1, wherein the nitrogen fixation and the photosynthesis occur contemporaneously.

3. The method of claim 1, wherein the bacteria includes cyanobacteria.

4. The method of claim 1 , performed without feeding bioreactor a sugar or any other chemical energy stream.

5. The method of claim 1, wherein the bioreactor facilitates digestion of the resulting biomass by anaerobic digestion, and separation and treatment the Anaerobic effluent from the bioreactor includes bio-fertilizer.

6. A method for producing a fertilizer product comprising: in a first vessel, providing a liquid solution comprising: water as a hydrogen source; a bacteria culture, the bacteria culture in an amount sufficient such that when activated, performs Nitrogen fixation and photosynthesis; nutrients for the bacteria culture; air as a Nitrogen source for Nitrogen fixation by the bacteria culture, and an oxygen source; and

Carbon Dioxide (CO₂) as a gas; exposing the liquid solution in the first vessel to light, including sunlight, to activate the bacteria culture and grow the bacteria in the bacteria culture; and removing at least a portion of the bacteria culture as biomass; in a second vessel, anaerobically digesting the bacteria culture of the biomass by reacting the biomass with an amount of anaerobic bacteria sufficient to produce Ammonium (NH4) in an effluent.

7. The method of claim 6, additionally comprising: converting the Ammonium of the effluent, in a third vessel, to a Nitrate by reacting the Ammonium with a Carbonate.

8. The method of claim 6, wherein the first vessel includes a photobioreactor.

9. The method of claim 8, wherein the photobioreactor includes one or more columns.

10. The method of claim 6, wherein the bacteria culture includes a cyanobacteria culture.

11. The method of claim 10, wherein the cyanobacteria culture includes cyanobacteria selected from the group consisting of: Anabaena flos aqua, Anabaena siamensis, Anabaena azollae, Anabaena variabilis, and mutant strains thereof.

12. The method of claim 10, wherein the cyanobacteria culture includes cyanobacteria of the family Nostocacae.

13. The method of claim 6, wherein the air includes ambient air.

14. The method of claim 6, wherein the nutrients include micro nutrients.

15. The method of clam 14, wherein the micronutrients include one or more of: Boron, Copper, Sodium, and Phosphorous.

16. The method of claim 6, wherein the nutrients include macronutrients.

17. The method of claim 6, wherein the macronutrients include one or more of: Magnesium, Potassium, and Calcium.

18. The method of claim 6, wherein the providing the Carbon Dioxide as a gas is in a gas stream at pressures sufficient to cause airlift in the liquid solution in the first vessel.

19. The method of claim 6, wherein the removing at least a portion of the bacteria culture as biomass is performed when the density of the bacteria culture reaches a predetermined value.

20. The method of claim 19, wherein the predetermined value is at least between 1 to 2 grams per liter.

21. The method of claim 6, wherein the removing at least a portion of the bacteria culture as biomass, includes determining the amount of the bacteria culture to be removed by a Machine Learning model.

22. The method of claim 6, wherein the removing at least a portion of the bacteria culture as biomass, includes determining the amount of the bacteria culture to be removed by a computer algorithm.

23. The method of claim 6, wherein the second vessel includes an anaerobic digester.

24. The method of claim 6, wherein the anaerobic bacteria for anaerobically digesting the biomass includes one or more of: protozoa, fungi, Cellulolytic Bacteria, Starch-Digesting Bacteria, Proteolytic Bacteria, Methanogenic Archaea, Lactic Acid Bacteria, and Sulfate-Reducing Bacteria.

25. The method of claim 6, wherein the bacteria culture is provided to the liquid solution as a starter culture in an inoculum charge.

26. The method of clam 7, wherein the third vessel includes a Nitrification bioreactor.

27. The method of clam 7, wherein the converting the Ammonium of the effluent to a Nitrate additionally comprises reacting the Ammonium effluent with a sufficient amount of aerobic bacteria.

28. The method of claim 27, wherein the aerobic bacteria is selected from the group consisting of: Nitrosomonas, Nitrosospira, Nitrosococcus, and Nitrosolobus.

29. The method of claim 27, wherein the aerobic bacteria include bacteria is of the family Nitrobacteraceae.

30. The method of claim 7, wherein the Carbonate includes one or more of: Calcium Carbonate, Potassium Carbonate, and Magnesium Carbonate.

31. The method of claim 30, wherein the Nitrate includes Calcium Nitrate, Potassium Nitrate, and/or Magnesium Nitrate.

32. A system for producing a fertilizer product comprising: a photobioreactor system comprising: a photobioreactor for growing a cyanobacteria culture using ambient nitrogen; and a harvesting unit in communication with the photobioreactor for harvesting the cyanobacteria when grown as biomass; and a fertilizer system comprising: an anaerobic digestion bioreactor for converting harvested biomass to Ammonium (NH4) in an effluent; and a Nitrification bioreactor in communication with the anaerobic digester for converting the Ammonium in the effluent to Ammonium Nitrate.

33. The system of claim 32, additionally comprising a controller in communication with sensors in the photobioreactor, the controller for 1) determining the density of the cyanobacteria culture in the photobioreactor, and 2) based on the density determination, determining whether to harvest at least a portion of the cyanobacteria culture.

34. The system of claim 33, wherein the controller applies at last one of a Machine Learning model or a computer algorithm, to determine the time to harvest the at last a portion of the cyanobacteria culture and the amount of the at least a portion of the cyanobacteria culture to be harvested.

35. The system of claim 32, wherein the harvesting unit includes a centrifuge and/or a membrane filtration system.

36. The system of claim 32, wherein the photobioreactor includes one or more columns of a material translucent or transparent to sunlight.

37. A method for producing a fertilizer product comprising: providing a liquid solution comprising: water as a hydrogen source; a cyanobacteria culture, the cyanobacteria culture in an amount sufficient such that when activated, performs Nitrogen fixation and photosynthesis; nutrients for the bacteria culture; ambient air as a Nitrogen source for Nitrogen fixation by the bacteria culture, and an oxygen source; and

Carbon Dioxide (CO₂) as a gas; and exposing the liquid solution in the first vessel to light, including sunlight, to activate the cyanobacteria culture and grow the bacteria in the bacteria culture.