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WO2025036972A1 - Produits protéiques monocellulaires contenant de faibles niveaux d'acides nucléiques - Google Patents

Produits protéiques monocellulaires contenant de faibles niveaux d'acides nucléiques Download PDF

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Publication number
WO2025036972A1
WO2025036972A1 PCT/EP2024/073003 EP2024073003W WO2025036972A1 WO 2025036972 A1 WO2025036972 A1 WO 2025036972A1 EP 2024073003 W EP2024073003 W EP 2024073003W WO 2025036972 A1 WO2025036972 A1 WO 2025036972A1
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WIPO (PCT)
Prior art keywords
yeast
composition
scp
product
fatty acids
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English (en)
Inventor
Willem Aike HOMMES
Mickel Leonardus August JANSEN
Wouter KROES
Evert Tjeerd VAN RIJ
Andre VENTE
Koen Johannes Anthonius VERHAGEN
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DSM IP Assets BV
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DSM IP Assets BV
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Publication of WO2025036972A1 publication Critical patent/WO2025036972A1/fr
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/142—Amino acids; Derivatives thereof
    • A23K20/147—Polymeric derivatives, e.g. peptides or proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/153—Nucleic acids; Hydrolysis products or derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/40—Meat products; Meat meal; Preparation or treatment thereof containing additives
    • A23L13/42—Additives other than enzymes or microorganisms in meat products or meat meals
    • A23L13/424—Addition of non-meat animal protein material, e.g. blood, egg, dairy products, fish; Proteins from microorganisms, yeasts or fungi

Definitions

  • the present invention relates to a composition
  • a composition comprising yeast single cell protein (SCP) product comprising 6% or less (w/w) nucleic acid content per gram of dry yeast SCP product.
  • the yeast single cell protein product is produced utilizing ethanol or molasses as a feedstock.
  • the yeast single cell protein product contains advantageously high levels of unsaturated versus saturated fatty acids.
  • SCPs single cell proteins
  • animals feed e.g. for poultry, calves, pigs, and fish
  • SCP product production still faces several challenges, like availability and sustainability of feedstock, like waste-based feedstock.
  • feed millers and farmers still require alternative solutions for protein production to be able to provide low carbon footprint diets while ensuring an environmental-friendly use of the planet's resources.
  • the present invention aims at providing a composition comprising yeast single cell protein (SCP) product comprising 6% or less (w/w) nucleic acid content per gram of dry yeast SCP product, thus providing a nutritionally rich alternative protein source that can supplement or even replace currently used animal- and/or plant-derived protein sources in animal feed.
  • SCP yeast single cell protein
  • said yeast single cell protein (SCP) product comprises 5% or less (w/w) nucleic acid content per gram of dry yeast SCP product.
  • said yeast single cell protein (SCP) product comprises 4% or less (w/w) nucleic acid content per gram of dry yeast SCP product.
  • said nucleic acid content comprises nucleosides and/or nucleotides.
  • said nucleosides are ribonucleosides and/or deoxyribonucleosides.
  • nucleotides are ribonucleotides and/or deoxyribonucleotides.
  • yeast SCP product further comprises crude protein.
  • yeast SCP product further comprises carbohydrates.
  • said yeast SCP product further comprises crude lipids.
  • said yeast SCP product further comprises crude protein, carbohydrates and crude lipids.
  • said yeast single cell protein (SCP) product comprises 35%, 40%, 45%, or 50% or more (w/w) crude protein per gram of dry yeast SCP product.
  • said yeast SCP product comprises at least about 14% (w/w) essential amino acids per gram of dry yeast SCP product.
  • said yeast SCP product comprises at least about 18% (w/w) essential amino acids per gram of dry yeast SCP product.
  • said yeast single cell protein (SCP) product comprises 40%, 30%, 25% or 20% or less (w/w) total carbohydrates per gram of dry yeast SCP product.
  • said yeast SCP product comprises about 10% or less (w/w) crude lipids per gram of dry yeast SCP product.
  • said yeast single cell protein (SCP) product comprises about 8% or less (w/w) crude lipids per gram of dry yeast SCP product.
  • said yeast single cell protein (SCP) product comprises about 6% or less (w/w) crude lipids per gram of dry yeast SCP product.
  • said yeast single cell protein (SCP) product comprises between about 10% to 8%, 10% to 6%, 10% to 4%, 8% to 6%, 8% to 4%, 6% to 4% (w/w) crude lipids per gram of dry yeast SCP product.
  • said yeast single cell protein (SCP) product comprises crude lipids to carbohydrates in a ratio of 0.6 to 1 : 2 to 5.
  • said yeast single cell protein (SCP) product comprises crude lipids to crude protein in a ratio of 0.6 to 1 : 3.5 to 6.
  • said yeast single cell protein (SCP) product comprises crude lipids, carbohydrates and crude protein in a ratio of 0.6 to 1 crude lipids : 2 to 5 carbohydrates : 3.5 to 6 crude protein.
  • said crude lipids comprise saturated fatty acids, mono-unsaturated fatty acids and poly-unsaturated fatty acids.
  • said yeast single cell protein (SCP) product comprises unsaturated fatty acids, preferably C16:1, C18:1, C18:2, and C18:3, and saturated fatty acids, preferably C16:0 and C18:0, in a ratio equal to or greater than 1.0.
  • said yeast single cell protein (SCP) product comprises unsaturated fatty acids, preferably C16:1, C18:1, C18:2, and C18:3, and saturated fatty acids, preferably C16:0 and C18:0, in a ratio equal to or greater than 1.1 , preferably greater than 1.2, more preferably greater than 1.5, most preferably greater than 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, or 8.0.
  • said saturated fatty acids are C16:0 and C18:0 fatty acids.
  • said C16:0 fatty acids and C18:0 fatty acids are present in an amount of no more than 0.85% (w/w) per gram of dry yeast SCP product, preferably 0.53% (w/w) per gram of dry yeast SCP product.
  • said mono-unsaturated fatty acids are C16:1 fatty acids and C18:1 fatty acids.
  • said C16:1 fatty acids and C18:1 fatty acids are present in an amount of at least about 2.3% (w/w) per gram of dry yeast SCP product, preferably 2.8% (w/w) per gram of dry yeast SCP product.
  • said poly-unsaturated fatty acids are C18:2 and C18:3 fatty acids.
  • said C18:2 fatty acids and C18:3 fatty acids are present in an amount of at least about 1.0% (w/w) per gram of dry yeast SCP product, preferably 2.0% (w/w) per gram of dry yeast SCP product.
  • said yeast SCP product is obtained from cultivating a Saccharomycetales yeast.
  • Saccharomycetales yeast is a yeast from the genus Cyberlindnera, Saccharomyces, Kluyveromyces, Wickerhamomyces, Pichia or Yarrowia, preferably from the genus Cyberlindnera or Saccharomyces or Kluyveromyces or Wickerhamomyces.
  • Saccharomycetales yeast is a yeast from Cyberlindnera jadinii, Saccharomyces cerevisiae, Kluyveromyces lactis, Wickerhamomyces anomalus, Pichia anomala or Yarrowia lipolytica, preferably from Cyberlindnera jadinii or Saccharomyces cerevisiae or Kluyveromyces lactis or Wickerhamomyces anomalus.
  • Saccharomycetales yeast is a yeast from Cyberlindnera jadinii ATCC 26387, Cyberlindnera jadinii FERM-BP1656, Cyberlindnera jadinii CBS621 , Cyberlindnera jadinii CBS841, Saccharomyces cerevisiae GHP1, Saccharomyces cerevisiae CEN.PK113-7D, Wickerhamomyces anomalus IFO 569, Wickerhamomyces anomalus CBS 1980, Cyberlindnera jadinii ATCC 9950, Kluyveromyces lactis CBS 2896, Wickerhamomyces anomalus CBS 2576 or Yarrowia lipolytica CBS 7504.
  • the term “derived from” preferably refers to yeast cells, which were originally obtained from a given yeast strain and thus originate from said given yeast strain. Such derived cells may differ from said given yeast strain due to naturally occurring and/or artificially introduced alterations like genetic mutations, but preferably have similar characteristics as cells from the yeast strain they originated from. Accordingly, cells that are derived from a given strain may have, preferably on genome level, a sequence identity of 80% or more, preferably of 85% or more, more preferably of 90% or more, even more preferably of 95% or more to the respective strain that can be seen as reference.
  • a derived cell may have a sequence identity of at least, e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% to the respective reference, preferably on genome level.
  • said Saccharomycetales yeast is cultivated by feeding ethanol, glucose or molasses as feedstock, with ethanol being preferred.
  • the growth rate of said Saccharomycetales yeast is below 0.2 h’ 1 , preferably below 0.15 h’ 1 , more preferably below 0.1 h’ 1 , more preferably below 0.05 h’ 1 , more preferably the growth rate is between 0.02 and 0.05 h’ 1 , more preferably between 0.025 and 0.05 h’ 1 , most preferably between 0.025 and 0.045 h’ 1 .
  • the feed rate is 0.179 - 0.536 g e thanoi/gbiomass/h, more preferably 0.179 - 0.469 g et hanoi/gbiomass/h.
  • the temperature for cultivating said Saccharomycetales yeast is kept between 30 and 40°C, preferably between 30 and 38°C, more preferably between 30 and 36°C, most preferably between 30 and 34°C.
  • the pH for cultivating said Saccharomycetales yeast is kept between 3.5 and 5.5, preferably between 3.5 and 5.5, more preferably between 3.5 and 4.5.
  • said composition is animal feed or human food or ingredient of human food, e.g. as meat analogue.
  • the composition of the invention is a yeast single cell protein (SCP) product that comprises low levels of nucleic acids per gram of dry yeast SCP product.
  • Saccharomycetales yeast cells when grown with e.g. ethanol as a carbon source, produce low levels of nucleic acid content, i.e. 6% (w/w) or less nucleic acid content per gram dry weight of the yeast SCP product produced, preferably 5% (w/w) or less essential amino acid per gram dry weight of the yeast SCP product produced, more preferably 4% (w/w) or less essential amino acid per gram dry weight of the yeast SCP product produced.
  • Saccharomycetales yeast cells are preferably from the genus Cyberlindnera or Kluyveromyces or Wickerhamomyces, and more preferably the Saccharomycetales yeast cells are from the genus Cyberlindnera jadinii or Kluyveromyces lactis or Wickerhamomyces anomalus.
  • Saccharomycetales yeast cells appear to produce less nucleic acids when grown with ethanol as carbon source.
  • Saccharomycetales yeast cells appear to be suitable as single cell protein product, with the aim of at least partially, preferably fully replacing animal-derived protein sources, such as fish meal in animal feed.
  • Saccharomycetales yeast cells can be grown by utilizing any carbon source, such as molasses, glucose or ethanol, with ethanol being preferred.
  • the term “”w/w” is intended to be understood as "weight by weight” and thus refers to the proportion of a particular substance within a mixture, as measured by weight or mass.
  • the nucleic acid content may be quantified by any appropriate method known to those skilled in the art. Examples include quantitative PCR (qPCR), digital PCR (dPCR), and UV spectrophotometry.
  • qPCR quantitative PCR
  • dPCR digital PCR
  • UV spectrophotometry UV spectrophotometry
  • nucleic acid preferably refers to a biomolecule compose of polymerized nucleotides, which comprise a pentose sugar, a phosphate group, and a nitrogenous base.
  • Biologically occurring nucleotides consist of ribonucleotides (RNA) and deoxyribonucleotides (DNA).
  • said nucleic acid content comprises nucleosides and/or nucleotides, wherein it is preferred that said nucleosides are ribonucleosides and/or deoxyribonucleosides, and wherein it is preferred that said nucleotides are ribonucleotides and/or deoxyribonucleotides.
  • the present invention relates to a composition comprising yeast single cell protein (SCP) product comprising at most about 6% (w/w) nucleic acid content per gram of dry yeast SCP product.
  • SCP yeast single cell protein
  • single cell protein refers to a protein obtained by and/or derived from a (unicellular) microorganism.
  • an SCP may refer to a protein purified and/or isolated from a microorganism’s cell culture for example.
  • SCPs may refer to microbial proteins are the dead dried cells of microorganisms.
  • an “single cell protein product” or “SCP product” may or may not comprise one or more selected from the group of intact (unicellular) microorganism cells, disrupted (unicellular) microorganism cells, isolated proteins obtained by one or more (unicellular) microorganism(s), isolated proteins derived by one or more (unicellular) microorganism(s), purified proteins obtained by one or more (unicellular) microorganism(s), and purified proteins derived by one or more (unicellular) microorganism(s).
  • an (unicellular) microorganism may relate to a bacterium, a fungus like yeast and/or an algae
  • said (unicellular) microorganism is yeast according to the present invention.
  • SCP products from yeast offer the advantage of providing comparatively high protein contents, while at the same time said products can be produced on industrial scale at comparatively low cost, independent from seasonal effects and with comparatively low harvesting efforts. Thus, yeast SCP products are highly advantageous.
  • yeast refers to a eukaryotic, unicellular microorganism classified as a member of the fungus kingdom that mostly reproduce asexually by mitosis with asymmetric division processes also being known as budding.
  • said term preferably relates to yeast cells, which can be grown under artificial and/or lab conditions, e.g. as in vitro culture conditions, and in particular under standard laboratory conditions. Said term preferably also embraces yeast cells of a single type that have been grown in the laboratory for several generations and thus, said term preferably embraces also potential mutants of a yeast cell and/or strain.
  • yeast is preferably Saccharomycetales yeast.
  • mutant means permanent (epi-) genetic modification(s) of genetic material, i. e. nucleic acids, caused, for example, naturally or by physical means or chemical compounds/substances/agents such as EMS.
  • Said modifications include point mutations, transitions, transversions, deletion/insertion/addition of one or more bases within a nucleic acid/gene/chromosome thereby optionally modifying the nucleic acid/gene/chromosome which can cause, inter alia, phenotypic effects like varying protein per dry matter (%) (w/w).
  • modification(s) may be induced by methods known to the person skilled in the art.
  • the skilled person is also aware of suitable methods to select cells in view of one or more favorable and/or desired phenotypic trait(s) like an increase of protein per dry matter (%) (w/w) and/or utilization of ethanol or molasses as carbon source.
  • the Saccharomycetales yeast cells are not genetically engineered.
  • the animal feed according to the present invention comprises up to 20% (w/w) or up to 10% (w/w) yeast SCP product, wherein the yeast SCP product comprises Saccharomycetales yeast cells, wherein said Saccharomycetales yeast cells are preferably not genetically engineered.
  • the yeast SCP product comprises Saccharomycetales yeast cells, wherein said Saccharomycetales yeast cells are preferably not genetically engineered.
  • This is advantageous to ensure that the SCP product comprised in the animal feed comprises only well-defined and/or well characterized yeast cells but no mutated an thus, potentially undefined and/or uncharacterized yeast cells. This is also advantageous for a constant SCP product quality.
  • genetic engineering is used in its broadest sense for methods known to the person skilled in the art to modify desired nucleic acids in vitro and in vivo , e.g. by targeted mutagenesis and/or recombinant DNA technology. Accordingly, said methods may comprise cloning, sequencing and transformation of recombinant nucleic acids, and appropriate vectors, primers, enzymes, host cells and the like are known by the skilled artisan.
  • genetically engineered cells are genetically engineered in view of high protein per dry matter (%), suitable essential amino acid composition, efficient ethanol or molasses usage as carbon source and the like in the context of the present invention.
  • a “yeast cell” is a cell of a yeast, preferably a cell of a yeast as described herein. Also included by said term are parts of a yeast cell. Such “parts” might refer to substances derived from a yeast cell. For example, when disrupted yeast cells are used, there might be yeast protein as well as yeast cell wall material.
  • Saccharomycetales refers to the order Saccharomycetales within the phylum Ascomycota. Members of Saccharomycetales are also known and sometimes referred to as budding yeasts.
  • SCP product producer may vary in their ability to use and/or utilize ethanol or molasses as carbon source for SCP production.
  • the yeast SCP product preferably comprises Saccharomycetales yeast cells, wherein said Saccharomycetales yeast cells are Saccharomycetales yeast cells from one or more Saccharomycetales yeast genera, species and/or strains that are capable of using ethanol as carbon source.
  • the Saccharomycetales yeast cells may be Saccharomycetales yeast cells from one or more genera selected from the group consisting of Cyberlindnera, Kluyveromyces, Wickerhamomyces, Yarrowia, Pichia and Saccharomyces.
  • the yeast SCP product comprises Saccharomycetales yeast cells, wherein said Saccharomycetales yeast is a yeast from Cyberlindnera jadinii, Saccharomyces cerevisiae, Kluyveromyces lactis, Wickerhamomyces anomalus, Pichia anomala or Yarrowia lipolytica, preferably from Cyberlindnera jadinii or Saccharomyces cerevisiae or Kluyveromyces lactis or Wickerhamomyces anomalus, preferably wherein said Saccharomycetales yeast is a yeast from Cyberlindnera jadinii ATCC 26387, Cyberlindnera jadinii FERM-BP1656, Cyberlindnera jadinii CBS621, Cyberlindnera jadinii CBS841, Saccharomyces cerevisiae GHP1, Saccharomyces cerevisiae CEN.PK113-7D, Wickerhamomyces anomalus IFO 569, Wickerhamomyces anomalus CBS
  • the composition comprising yeast single cell protein (SCP) product contains less than 14% nucleic acid content per gram of dry yeast SCP product, wherein the yeast SCP product comprises Saccharomycetales yeast cells, and wherein Saccharomycetales yeast cells are yeast cells from the genus Cyberlindnera, Saccharomyces, Kluyveromyces, Wickerhamomyces, Pichia or Yarrowia, preferably from the genus Cyberlindnera or Saccharomyces or Kluyveromyces or Wickerhamomyces.
  • yeast cells from said genera are capable of growing on a culture medium comprising ethanol as a carbon source.
  • the composition comprising yeast SCP product comprises 35%, 40%, 45%, or 50% or more (w/w) crude protein per gram of dry yeast SCP product.
  • the composition comprising yeast SCP product comprises at least about 14% (w/w) essential amino acids per gram of dry yeast SCP product, more preferably at least about 18% (w/w) essential amino acids per gram of dry yeast SCP product.
  • essential amino acid preferably refers to amino acids that cannot be synthesized by an animal from metabolic intermediates. Thus, such amino acids have to be supplied from an exogenous diet as they are required, e.g., for growth.
  • nine amino acids are considered essential: phenylalanine, valine, tryptophan, threonine, isoleucine, methionine, histidine, leucine, and lysine.
  • said nine essential amino acids are obtainable by a single complete protein containing all the essential amino acids.
  • Such complete proteins can be derived from animal-based sources of nutrition, whereas plant-based foods represent commonly a source for essential amino acids in the form of incomplete proteins.
  • the composition comprising yeast SCP product comprises 40%, 30%, 25% or 20% or less (w/w) total carbohydrates per gram of dry yeast SCP product.
  • the composition comprising yeast SCP product comprises about 10% or less (w/w) crude lipids per gram of dry yeast SCP product.
  • said crude lipids comprise, for example, about 10% to 8%, 10% to 6%, 10% to 4%, 8% to 6%, 8% to 4%, 6% to 4% (w/w) crude lipids per gram of dry yeast SCP product.
  • said crude lipids may comprise saturated fatty acids, mono-unsaturated fatty acids and poly-unsaturated fatty acids [0067] Additionally, it was also surprisingly found that yeast cells, when grown with e.g.
  • the ratio of unsaturated fatty acids preferably C16: 1 , C18:1 , C18:2 and C18:3, and saturated fatty acids, preferably C16:0 and C18:0, should be greater or equal to 1.0, preferably greater than 1.1, more preferably greater than 1.2, more preferably greater than 1.5, most preferably greater than 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, or 8.0.
  • the term “unsaturated fatty acid” preferably refers to a fatty acid in which there is at least one double bond within the fatty acid chain.
  • a fatty acid is “monounsaturated” if it contains a single double bond within the fatty acid chain and “polyunsaturated” if it contains more than one double bond within the fatty acid chain.
  • the potential health benefits of consuming unsaturated fats, more commonly found in plant material, over saturated fat, more commonly found in animal products may include, but are not limited to, lower risk of cardiovascular disease, stroke, type 2 diabetes, and certain types of cancer. The evidence to support those benefits varies across specific diseases, but the overall beneficial effect of increased unsaturated fat consumption is widely acknowledged.
  • the composition comprising yeast SCP product comprises crude lipids to carbohydrates in a ratio of 0.6 to 1 : 2 to 5 and/or comprises crude lipids to crude protein in a ratio of 0.6 to 1 : 3.5 to 6.
  • the saturated fatty acids of the yeast SCP product comprise C16:0 and C18:0 fatty acids, wherein preferably the C16:0 fatty acids and C18:0 fatty acids are present in an amount of no more than at least about 0.85% (w/w) per gram of dry yeast SCP product, preferably 0.53% (w/w) per gram of dry yeast SCP product.
  • the mono-unsaturated fatty acids of the yeast SCP product comprise C16:1 and C18:1 fatty acids, wherein preferably the C16:1 fatty acids and C18:1 fatty acids are present in an amount of no more than at least about 2.3% (w/w) per gram of dry yeast SCP product, preferably about 2.8% (w/w) per gram of dry yeast SCP product.
  • the poly-unsaturated fatty acids of the yeast SCP product comprise C16:2 and C18:2 fatty acids, wherein preferably the C16:2 fatty acids and C18:2 fatty acids are present in an amount of no more than at least about 1.0 % (w/w) per gram of dry yeast SCP product, preferably about 2.0% (w/w) per gram of dry yeast SCP product.
  • Saccharomycetales yeast is cultivated by feeding ethanol as feedstock.
  • the ethanol concentration inside the fermenter itself is always close to zero ( ⁇ 0.1 g/L), as any ethanol that is fed is virtually immediately consumed by the yeast.
  • the main parameter of the fermentation process that determines the growth rate, is the ethanol feed rate. This feed rate has to be started low, as the biomass concentration at the start of fermentation is low and is subsequently increased exponentially to match the exponential growth rate of the biomass. At a certain point during the fermentation, this exponentially increasing feed rate is fixed to a constant feed rate, as the supply of oxygen in the fermenter becomes limiting.
  • the maximum feed rate is limited by two main parameters in the fermentation process: 1) the biomass concentration, and 2) the oxygen transfer capacity.
  • the ethanol feed rate should be defined by the substrate uptake rate of the organism (the qs) calculated by the amount of substrate that is consumed (g e thanoi) per amount of biomass in the fermenter (gbiomass) per hour, of 0.179 - 0.536 g e thanoi/gbiomass/h, more preferably 0.179 - 0.469 g et hanoi/gbiomass/h.
  • the biomass yield decreases with higher temperatures, causing the protein yield (the amount of protein produced per gram of ethanol or molasses) to decrease with increasing temperatures.
  • the temperature is kept between 30 and 40°C, preferably between 30 and 38°C, more preferably between 30 and 36°C, most preferably between 30 and 34°C.
  • Protein content does change with the pH level. Protein content is increased at lower pH levels. The biomass yield however drops significantly at pH levels of 3.5 and lower. Preferably, the pH is kept between 3.5 and 5.5, preferably between 3.5 and 5.0, more preferably between 3.5 and 4.5.
  • the method for producing biomass may further comprise a step of recovering the biomass from the aerobic fermentation by suitable methods known in the art.
  • Recovering biomass may comprise centrifugation or filtration.
  • the method for producing biomass may further comprise a step of drying the biomass by suitable methods known in the art.
  • Drying biomass may comprise convective/direct drying technologies (like spray drying, fluidized bed) or contact/indirect technologies (like drum drying, vacuum drying, falling film) or supercritical drying (using superheated steam) or natural air/sun drying or even freeze drying.
  • composition of the present invention may be used as animal feed or as human food or ingredient of human food, e.g. as meat analogue.
  • animal feed refers to any compound, preparation, or mixture suitable for, or intended for intake by an animal (e.g., a fish).
  • Animal feed for a monogastric animal typically comprises concentrates as well as vitamins, minerals, enzymes, direct fed microbial, amino acids and/or other feed ingredients (such as in a premix)
  • animal feed for ruminants generally comprises forage (including roughage and silage) and may further comprise concentrates as well as vitamins, minerals, enzymes direct fed microbial, amino acid and/or other feed ingredients (such as in a premix).
  • An animal feed additive e.g., fish feed additive
  • a formulated enzyme product which may further comprise e.g. vitamins, minerals, enzymes, amino acids, preservatives and/or antibiotics; i.e. a premix.
  • the animal feed additive/premix is typically mixed in a feed mill with concentrates and/or forage such as vegetable protein, legumes or other plant material. Further, the animal feed is typically fed as a pelleted feed to monogastric animals.
  • said animal feed is preferably a feed for aquatic species.
  • said species are preferably selected from crustaceans or fish.
  • the animal feed may be a feed for crustaceans and/or fish.
  • said crustaceans are preferably shrimps.
  • said fish may preferably be warm water fish or cold water fish.
  • said fish are preferably selected from the group consisting of catfish, tilapia, seabream, seabass, and carp.
  • said fish are cold water fish with said fish being selected from cod, salmon, or rainbow trout.
  • the animal feed according to the present invention is a feed for shrimps, salmon and/or rainbow trout.
  • the uses of the present invention are non-therapeutic. Indeed, the improvement of the weight of an animal like of aquatic species is deemed to be associated and/or to contribute to, e.g. health or growth of the animal, which is by way of common sense non-therapeutic.
  • a composition comprising yeast single cell protein (SCP) product comprising 6% or less (w/w) nucleic acid content per gram of dry yeast SCP product.
  • SCP yeast single cell protein
  • composition of item 1 wherein said yeast single cell protein (SCP) product comprises 5% or less (w/w) nucleic acid content per gram of dry yeast SCP product.
  • SCP yeast single cell protein
  • composition of any one of the preceding items, wherein said yeast single cell protein (SCP) product comprises 4% or less (w/w) nucleic acid content per gram of dry yeast SCP product.
  • composition of any one of the preceding items, wherein said nucleic acid content comprises nucleosides and/or nucleotides.
  • composition of item 4 wherein said nucleosides are ribonucleosides and/or deoxyribonucleosides.
  • composition of item 4 wherein said nucleotides are ribonucleotides and/or deoxyribonucleotides.
  • composition of any one of the preceding items, wherein said yeast SCP product further comprises crude lipids. 10. The composition of any one of the preceding items, wherein said yeast SCP product further comprises crude protein, carbohydrates and crude lipids.
  • yeast single cell protein (SCP) product comprises 35%, 40%, 45%, or 50% or more (w/w) crude protein per gram of dry yeast SCP product.
  • composition of any one of the preceding items, wherein said yeast single cell protein (SCP) product comprises 40%, 30%, 25% or 20% or less (w/w) total carbohydrates per gram of dry yeast SCP product.
  • yeast single cell protein (SCP) product comprises about 8% or less (w/w) crude lipids per gram of dry yeast SCP product.
  • yeast single cell protein (SCP) product comprises about 6% or less (w/w) crude lipids per gram of dry yeast SCP product.
  • yeast single cell protein (SCP) product comprises between about 10% to 8%, 10% to 6%, 10% to 4%, 8% to 6%, 8% to 4%, 6% to 4% (w/w) crude lipids per gram of dry yeast SCP product.
  • yeast single cell protein (SCP) product comprises crude lipids to carbohydrates in a ratio of 0.6 to 1 : 2 to 5.
  • composition of any one of the preceding items, wherein said yeast single cell protein (SCP) product comprises crude lipids to crude protein in a ratio of 0.6 to 1 : 3.5 to 6.
  • the composition of any one of the preceding items, wherein said yeast single cell protein (SCP) product comprises crude lipids, carbohydrates and crude protein in a ratio of 0.6 to 1 crude lipids : 2 to 5 carbohydrates : 3.5 to 6 crude protein.
  • composition of any one of the preceding items, wherein said yeast single cell protein (SCP) product comprises unsaturated fatty acids, preferably C16:1, C18:1, C18:2 and C18:3 and saturated fatty acids, preferably C16:0 and C18:0 in a ratio equal to or greater than 1.0.
  • SCP yeast single cell protein
  • composition of item 23 wherein said yeast single cell protein (SCP) product comprises unsaturated fatty acids, preferably C16:1, C18:1, C18:2 and C18:3, and saturated fatty acids, preferably C16:0 and C18:0, in a ratio equal to or greater than 1.1 , preferably greater than 1.2, more preferably greater than 1.5, most preferably greater than 2.0, 3.0, 4.0, 5.0, 6.0, 7.0 or 8.0.
  • SCP yeast single cell protein
  • composition of item 23 wherein C16:0 fatty acids and C18:0 fatty acids are present in an amount of no more than 0.85% (w/w) per gram of dry yeast SCP product, preferably 0.53% (w/w) per gram of dry yeast SCP product.
  • composition of item 22 wherein said mono-unsaturated fatty acids are C16:1 fatty acids and C18:1 fatty acids.
  • composition of item 22 wherein said poly-unsaturated fatty acids are C18:2 and C18:3 fatty acids.
  • yeast SCP product is obtained from cultivating a Saccharomycetales yeast. 32.
  • composition of item 31 wherein the Saccharomycetales yeast is a yeast from the genus Cyberlindnera, Saccharomyces, Kluyveromyces, Wickerhamomyces, Pichia or Yarrowia, preferably from the genus Cyberlindnera or Saccharomyces or Kluyveromyces or Wickerhamomyces.
  • composition of item 31 or 32, wherein the Saccharomycetales yeast is a yeast from Cyberlindnera jadinii, Saccharomyces cerevisiae, Kluyveromyces lactis, Wickerhamomyces anomalus, Pichia anomala or Yarrowia lipolytica, preferably from Cyberlindnera jadinii or Saccharomyces cerevisiae or Kluyveromyces lactis or Wickerhamomyces anomalus.
  • Saccharomycetales yeast is a yeast from Cyberlindnera jadinii ATCC 26387, Cyberlindnera jadinii FERM- BP1656, Cyberlindnera jadinii CBS621, Cyberlindnera jadinii CBS841, Saccharomyces cerevisiae GHP1, Saccharomyces cerevisiae CEN.PK113-7D, Wickerhamomyces anomalus IFO 569, Wickerhamomyces anomalus CBS 1980, Cyberlindnera jadinii ATCC 9950, Kluyveromyces lactis CBS 2896, Wickerhamomyces anomalus CBS 2576 or Yarrowia lipolytica CBS 7504.
  • composition of any one of items 31 to 36, wherein the feed rate is 0.179 - 0.536 g e thanoi/gbiomass/h, more preferably 0.179 - 0.469 g et hanoi/gbiomass/h.
  • composition of any one of items 31 to 37, wherein the temperature for cultivating said Saccharomycetales yeast is kept between 30 and 40°C, preferably between 30 and 38°C, more preferably between 30 and 36°C, most preferably between 30 and 34°C.
  • composition of any one of items 31 to 38, wherein the pH for cultivating said Saccharomycetales yeast is kept between 3.5 and 5.5, preferably between 3.5 and 5.5, more preferably between 3.5 and 4.5.
  • composition of any one of the preceding items, wherein said composition is animal feed or human food or ingredient of human food, e.g. as meat analogue.
  • Figure 1 shows a table quantifying the content of the yeast SCP of the present invention, when grown with ethanol or molasses as carbon source. Shown are general parameters and broad categories, such as total carbohydrates, crude lipids, and crude proteins, along with a breakdown of specific amino acid and lipid content.
  • Example 1 SCP product content
  • SCP single cell protein
  • SCP product was generated using Saccharomycetales yeast cells using ethanol or molasses as carbon source.
  • Cyberlindnera jadinii was used.
  • Feed rate, growth rate and pH of the culture medium was chosen pursuant to the teaching of the present application.
  • the nutritional content of said SCP product was quantified, primarily as weight per total weight of dry product or % w/w of (dry) product. Content quantified includes total crude lipid content, total RNA/DNA content, amino acids, and fatty acids. These data indicate that the total nucleic acid content in the sample of SCP product was 6.0% as a percentage of dry product by weight. All measured values are shown in Table 1 , below.

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Abstract

La présente invention concerne une composition comprenant un produit de protéine monocellulaire de levure (SCP) comprenant au plus environ 6 % ou moins (p/p) de teneur en acide nucléique par gramme de produit SCP de levure sèche. De préférence, le produit protéique monocellulaire de levure est produit à l'aide d'éthanol ou de mélasse en tant que charge d'alimentation. Le produit protéique monocellulaire de levure contient avantageusement un rapport élevé d'acides gras insaturés à des acides gras saturés.
PCT/EP2024/073003 2023-08-16 2024-08-15 Produits protéiques monocellulaires contenant de faibles niveaux d'acides nucléiques Pending WO2025036972A1 (fr)

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EP23191800 2023-08-16
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EP23192197 2023-08-18
EP23192196 2023-08-18
EP23192196.6 2023-08-18
EP23192197.4 2023-08-18
EP23216997 2023-12-15
EP23216997.9 2023-12-15

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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA3203861A1 (fr) * 2021-02-17 2022-08-25 Hiroshi Kojo Aliment pour animal d`elevage

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA3203861A1 (fr) * 2021-02-17 2022-08-25 Hiroshi Kojo Aliment pour animal d`elevage

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Title
AGBOOLA JELEEL OPEYEMI ET AL: "Yeast as a novel protein source - Effect of species and autolysis on protein and amino acid digestibility in Atlantic salmon (Salmo salar)", AQUACULTURE, ELSEVIER, AMSTERDAM, NL, vol. 546, 10 August 2021 (2021-08-10), XP086820085, ISSN: 0044-8486, [retrieved on 20210810], DOI: 10.1016/J.AQUACULTURE.2021.737312 *
ANON: "Article 4 Request", 21 January 2021 (2021-01-21), XP093219331, Retrieved from the Internet <URL:https://food.ec.europa.eu/document/download/e6dfe844-b2cb-4fa1-8e63-e13c0fc4ef77_en?filename=novel-food_consult-status_torula-yeast.pdf> [retrieved on 20241029] *
CASTRO A. C. ET AL: "Reduction of Nucleic Acid Content in Candida Yeast Cells by Bovine Pancreatic Ribonuclease A Treatment", APPLIED MICROBIOLOGY, vol. 22, no. 3, 1 September 1971 (1971-09-01), US, pages 422 - 427, XP093218828, ISSN: 0003-6919, DOI: 10.1128/am.22.3.422-427.1971 *
HEE JEONG CHAE ET AL: "Utilization of brewer's yeast cells for the production of food-grade yeast extract. Part 1: effects of different enzymatic treatments on solid and protein recovery and flavor characteristics", vol. 76, no. 3, 1 February 2001 (2001-02-01), pages 253 - 258, XP009049996, ISSN: 0960-8524, Retrieved from the Internet <URL:https://www.sciencedirect.com/science/article/pii/S0960852400001024?via%3Dihub> [retrieved on 20001127], DOI: 10.1016/S0960-8524(00)00102-4 *
KOT ANNA M. ET AL: "Magnesium Binding by Cyberlindnera jadinii Yeast in Media from Potato Wastewater and Glycerol", MICROORGANISMS, vol. 11, no. 8, 28 July 2023 (2023-07-28), Switzerland, pages 1923, XP093218832, ISSN: 2076-2607, DOI: 10.3390/microorganisms11081923 *

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