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WO2015083769A1 - Procédé de production de polyhydroalcanoate utilisant uniquement la photosynthèse - Google Patents

Procédé de production de polyhydroalcanoate utilisant uniquement la photosynthèse Download PDF

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Publication number
WO2015083769A1
WO2015083769A1 PCT/JP2014/082073 JP2014082073W WO2015083769A1 WO 2015083769 A1 WO2015083769 A1 WO 2015083769A1 JP 2014082073 W JP2014082073 W JP 2014082073W WO 2015083769 A1 WO2015083769 A1 WO 2015083769A1
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pha
algae
acetoacetyl
cyanobacteria
gene
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Japanese (ja)
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松井　南
志夫 栗原
ニョクシン ラウ
フーンチュン ピン
スーディッシュ クマール
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RIKEN
Universiti Sains Malaysia (USM)
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RIKEN
Universiti Sains Malaysia (USM)
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/93—Ligases (6)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/12—Unicellular algae; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/62—Carboxylic acid esters
    • C12P7/625—Polyesters of hydroxy carboxylic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
    • C12Y101/01036—Acetoacetyl-CoA reductase (1.1.1.36)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y602/00—Ligases forming carbon-sulfur bonds (6.2)
    • C12Y602/01—Acid-Thiol Ligases (6.2.1)
    • C12Y602/01016—Acetoacetate-CoA ligase (6.2.1.16)

Definitions

  • the present invention relates to an algae into which acetoacetyl-CoA synthase has been introduced and use of the algae in the production of polyhydroxyalkanoic acid (PHA).
  • PHA polyhydroxyalkanoic acid
  • Cyanobacteria are one of the oldest photosynthetic organisms on the planet and have played an important role in the evolution of the oxygen atmosphere we breathe today (Non-patent Document 1). Even today, cyanobacteria play a central role in global carbon recycling, the nitrogen cycle, and most importantly, the maintenance of atmospheric composition (Non-Patent Documents 2 and 3). Cyanobacteria are considered to be ideal producers of various chemicals and biofuels because they use solar energy to fix carbon dioxide to biomass. In the natural environment, fluctuations in nitrogen concentration occur constantly, and microorganisms respond to nutrient starvation by accumulating various carbon and energy storage compounds (Non-patent Document 4). Research on these storage polymers, especially PHA, has attracted considerable interest in recent years in an attempt to address the waste disposal problem caused by petrochemical plastics (Non-Patent Document 5).
  • Non-Patent Document 6 An interesting and promising approach is the use of photosynthetic cyanobacteria as a host for PHA production. Cyanobacteria as a “microbe factory” can directly fix carbon dioxide from the atmosphere to high molecular weight PHA by photosynthesis.
  • cyanobacteria In addition to being photoautotrophic, cyanobacteria require minimal nutrients for growth, thus reducing the cost of a growth medium that is complex with the carbon source (Non-Patent Document 7). Thus, the use of cyanobacteria provides a cost-effective and sustainable approach for the production of this environmentally friendly polymer.
  • Non-Patent Document 8 Since then, many studies have been conducted on Aphanothece sp. (Non-patent document 9), Oscillatoria limosa (Non-patent document 10), and several species of the genus Spirulina (Non-patent documents 11 and 12).
  • Non-Patent Document 13 cyanobacteria have been characterized by the ability to produce PHA containing only 3-hydroxybutyrate (3HB) and / or 3-hydroxyvalerate (3HV) monomers.
  • 3HB 3-hydroxybutyrate
  • HV 3-hydroxyvalerate
  • the present invention has been made in view of such circumstances, and an object thereof is to produce polyhydroxyalkanoic acid with high efficiency and low cost by using algae.
  • Algae can grow without the need for other nutrient sources by photosynthesis using CO 2 as a carbon source.
  • cyanobacteria (Cinecocity strain PCC6803) uses genomic information (Kaneko T et al. (1996) DNA Res 3: 109-136) and ease of genetic manipulation by natural transformation ability (Williams JGK ( 1988) Method Enzymol 167: 766-778), suitable for various biotechnology production. Therefore, the present inventor has attempted metabolic alterations in cyanobacteria by increasing the flow of intermediates to PHA biosynthesis and introducing a highly active PHA synthase.
  • ⁇ -ketothiolase (phaA; derived from Capriavidas), acetoacetyl-CoA reductase (phaB; derived from Capriavidas), and PHA polymerization enzyme (phaC A gene encoding three enzymes (from the genus Chromobacteria).
  • phaA ⁇ -ketothiolase
  • phaB acetoacetyl-CoA reductase
  • phaC PHA polymerization enzyme
  • the present invention relates to the algae into which the acetoacetyl-CoA synthase gene has been introduced and the use of the algae in the production of polyhydroxyalkanoic acid. More specifically, the present invention provides the following inventions.
  • a method for enhancing the ability to produce polyhydroxyalkanoic acid in algae including introducing acetoacetyl-CoA synthase into algae.
  • [8] A method for producing polyhydroxyalkanoic acid, comprising culturing the algae according to any one of [1] to [6] under conditions capable of photosynthesis.
  • Bioplastics are bio-derived plastics that are beginning to be used in various applications such as beverage containers, car interiors, and personal computers.
  • a material having degradability (biodegradability) by microorganisms has been developed, and an environmental load reduction effect that is not found in plastics derived from petroleum is expected.
  • its productivity is very low, such as several percent or less of the dry weight of the plant or microorganism used, and problems such as growth delay of the plant and microorganism have been reported.
  • the production of bioplastics requires a large amount of sugar and special facilities for culturing microorganisms, and there are problems in terms of cost.
  • the present invention by using an algae into which an acetoacetyl-CoA synthase gene has been introduced, together with an unexpected increase in photosynthetic activity, 14% of the dry weight of cyanobacteria without a solution carbon source, and further as a carbon source By adding a small amount of acetic acid, the world's highest level of PHA production efficiency of 41% was achieved. According to the present invention, it has become possible to dramatically increase the production efficiency of PHA in algae and to greatly reduce the production cost.
  • Synechocystis PCC6803 strain is a graph showing a comparison of accumulation of P (3HB) in (pTKP2031V, C Cs A Cn B Cn, and C Cs NphT7B Cn).
  • A those bubble 2-3% CO 2
  • B 0.4% (w / v) which was shaken supplemented with acetic acid
  • C air exchange limited conditions, 0.4% (w / v) It was replenished with acetic acid and shaken.
  • AP allophycocyanin
  • PC / PEC phycocyanin / phycoerythrocyanin
  • Cytb 6 / f cytochrome b6 / f complex
  • PQ plastoquinone FNR, ferredoxin-NADP (+) reductase
  • Pc plastocyanin
  • PSI photosystem I
  • PSII photosystem II
  • Ndh NADH dehydrogenase
  • Glc-6-P glucose-6-phosphate
  • Fru-6-P fructose-6-phosphate
  • Fru-1 , 6-bp fructose-1,6-bisphosphate
  • Glycerate-1,3-P 2 1,3-bisphosphoglycerate (1,3-biphosphoglycerate); 3- P-Glycerate, 3-phosphoglycerate
  • Ru-1,5-bisP ribulose-1,5-biphosphate
  • PEP phosphoenolpyruvate.
  • the present invention provides algae into which acetoacetyl-CoA synthase has been introduced.
  • “algae” means an organism having photosynthetic ability among organisms living in the hydrosphere.
  • Examples of the algae used in the present invention include cyanobacteria and marine algae. However, the growth rate is fast, the genetic recombination technique is established, the genome sequence is decoded, and the gene Cyanobacteria are preferable for the reason that expression vectors are maintained.
  • cyanobacteria include the genus Synechocystis, the genus Microcystis (for example, Microcystis aeruginosa), the genus Arthrospira (for example, Arthrospira platensis), Cyanothece genus cyanobacteria, Alcaligenes genus cyanobacterium (eg, Alcaligenes eurtophus), Anabaena genus cyanobacterium, Synechococcus genus cyanobacterium, Thermosynechococcus (Thermosynechococcus) (E.g., Thermosynechococcus elongats), Gloeobacter genus cyanobacterium (e.g., Gloeobacter violaceus), Acaryochris (Acaryoch) loris) cyanobacterium (for example, Acaryochloris marina), Nostoc genus cyanobacterium (
  • the “acetoacetyl CoA synthase” used in the present invention is an enzyme that catalyzes a reaction for producing acetoacetyl CoA by irreversible condensation of acetyl CoA and malonyl CoA.
  • the acetoacetyl CoA synthetase introduced into the algae is not particularly limited as long as it has the above enzyme activity, but nphT7 is preferable.
  • the nucleotide sequence and amino acid sequence of typical nphT7 derived from the genus Streptomyces are shown in SEQ ID NOs: 1 and 2, respectively.
  • acetoacetyl CoA reductase and PHA polymerizing enzyme can be further introduced into algae.
  • the “acetoacetyl CoA reductase” in the present invention is an enzyme that catalyzes a reaction for producing 3-hydroxybutyl CoA from acetoacetyl CoA.
  • the acetoacetyl CoA reductase introduced into the algae is not particularly limited as long as it has the above enzyme activity, but phaB is preferable.
  • the base sequence and amino acid sequence of typical phaB derived from the genus Capriavidas are shown in SEQ ID NOs: 3 and 4, respectively, and the base sequence and amino acid sequence of phaB from a typical Pseudomonas genus are shown in SEQ ID NOs: 5 and 6, respectively.
  • the “PHA polymerization enzyme” in the present invention is an enzyme that catalyzes a reaction for producing PHA from 3-hydroxybutyl CoA.
  • the PHA polymerase introduced into the algae is not particularly limited as long as it has the above enzyme activity, but phaC is preferable.
  • the base sequence and amino acid sequence of phaC derived from a typical genus Chromobacterium are shown in SEQ ID NOs: 7 and 8, respectively.
  • amino acid sequence of the above-mentioned enzyme introduced into algae may be mutated in nature, or may be artificially mutated.
  • an enzyme in which one or more amino acids for example, within 30 amino acids, within 10 amino acids, within 5 amino acids, within 3 amino acids
  • substitution, deletion, insertion, addition is also used in the present invention can do.
  • the phrase “introduced” into the algae means that the enzyme is introduced into the algae as a gene and includes the case where the enzyme is introduced into the algae as a protein.
  • the gene encoding the enzyme can be inserted into an appropriate vector and introduced into algae by a general method such as natural transformation, electroporation, or conjugation.
  • a promoter for expressing a gene for example, a light-inducible psbAII promoter can be used.
  • a microinjection method or a method of introducing it in combination with a signal peptide can be used.
  • the present invention provides a method for enhancing the ability to produce PHA in algae, which comprises introducing the enzyme into the algae.
  • PHA can be produced with high efficiency by culturing the algae with enhanced PHA production ability under conditions capable of photosynthesis. Therefore, the present invention also provides a method for producing PHA, which comprises culturing the algae into which the enzyme is introduced under conditions capable of photosynthesis.
  • the cultivation of the algae into which the enzyme has been introduced can be carried out in a medium of an inorganic salt containing no sugar, thereby reducing the cultivation cost.
  • a medium to be used for example, BG-11 medium or the like can be used.
  • a limited nutrient source for example, a BG-11 medium lacking sodium nitrate
  • the production efficiency of PHA can be further increased.
  • conditions for the pH of the medium, the culture temperature, and the illumination general conditions such as pH 8.0, 30 ° C., and 100 ⁇ mol photons m ⁇ 2 s ⁇ 1 can be used. In culture, it is preferable to perform shaking or bubble with air.
  • PHA production efficiency can be dramatically increased by culturing under the application of a carbon source.
  • Carbon dioxide or acetic acid can be used as the carbon source.
  • the bubbled air may be enriched with carbon dioxide.
  • the concentration of carbon dioxide is preferably 2 to 3% (v / v).
  • concentration of acetic acid to be added is preferably 0.4% (w / v) or more.
  • the production efficiency of PHA can be remarkably increased by setting the air exchange restriction condition.
  • the air exchange restriction condition can be created, for example, by closing the mouth of the incubator with a cotton plug and covering with an aluminum foil.
  • the recovery of PHA from the algae thus cultured can be performed, for example, by an extraction method using methanol or ethanol, or a method using a crushing device such as Beadsbeader.
  • the amount of PHA produced in algae can be quantified by subjecting the dried cells to chloroform extraction, decomposing the extract with methanol, and removing the organic phase containing hydroxyacylmethyl ester. It can be performed by subjecting to gas chromatography mass spectrometry.
  • Escherichia coli DH5 ⁇ used for plasmid cloning was grown in LB medium at 37 degrees with shaking (180 rpm). Kanamycin (50 ⁇ g / mL) or ampicillin (100 ⁇ g / mL) was added for selection and maintenance of the plasmid.
  • Kanamycin 50 ⁇ g / mL
  • ampicillin 100 ⁇ g / mL
  • a two-stage culture was performed. Cultures were first grown in BG-11 medium until late logarithmic growth phase, then harvested, washed and transferred to BG-11 medium lacking sodium nitrate. Phosphorus deficiency was performed by culturing cells in BG-11 without potassium phosphate.
  • the construct for the transformation of Synechocystis is derived from pTKP2031V.
  • pTKP2031V was designed for homologous recombination with the kanamycin resistance cassette into the genome between sites slr2030 and slr2031 (Satoh S et al. (2001) J Biol Chem 276: 4293-4297).
  • Expression of all cyanobacteria constructs was under the control of the psbAII promoter.
  • a gene cluster containing ⁇ -ketothiolase (phaA Cn ) and acetoacetyl CoA reductase (phaB Cn ) was transformed into C. nekatol H16 using primers phaAB Cn (F; NdeI) and phaAB Cn (R; HpaI) (Table 1). Amplified from chromosomal DNA.
  • the underline in the table indicates the restriction enzyme digestion site.
  • the obtained PCR product was digested with NdeI and HpaI and inserted into pTKP2031V digested with NdeI and HpaI to obtain pTKP2031V-phaAB Cn .
  • PHA synthase (phaC Cs ) was prepared from chromosomal DNA of Chromobacterium sp. USM2 using primers phaC Cs (F; SfuI) and phaC Cs (R; AatI). This PCR fragment was digested with SfuI and AatI and subcloned into an appropriate restriction enzyme site of pTKP2031V-phaAB Cn by ligation to obtain pTKP2031V-phaC Cs A Cn B Cn .
  • pTKP2031V-phaC Cs nphT7phaB Cn was constructed by replacing phaA Cn with nphT7 in pTKP2031V-phaC Cs A Cn B Cn .
  • the nphT7 gene derived from Streptomyces sp. 190 was prepared using the template pHis_nphT7 (Okamura E et al. (2010) Proc Natl Acad, which was prepared in advance using primers nphT7 (F; SfuI) and nphT7 (R; AatI). Sci USA 107: 11265-11270). The transformation of Synechocystis was performed according to a previous report (Osanai T et al.
  • the transcription level of the gene of interest was normalized to the level of the housekeeping gene (16S rRNA) used in this study.
  • Synechocystis strain PCC6803 strain C Cs NphT7B Cn and C Cs A Cn B Cn (target) expression level for pTKP2031V (reference material) and C Cs A Cn B Cn (reference material) C Cs NphT7B Cn (target) In order to compare the expression level of the gene of interest, comparative quantification was performed.
  • RNA-seq library preparation 2 ⁇ g of total RNA was subjected to ribosomal RNA removal using Ribo-Zero rRNA removal kit (Epicentre, USA).
  • a cDNA library for RNA-seq was constructed from total RNA from which rRNA was removed using the Illumina TruSeq Stranded mRNA Sample Preparation Kit (Illumina, USA) according to the manufacturer's instructions. Briefly, the preparation of a cDNA library includes the following steps. RNA fragmentation, cDNA synthesis, 3 ′ end adenylation, adapter ligation, and enrichment of cDNA templates.
  • the nphT7 Ss gene catalyzes the irreversible condensation of acetyl-CoA and malonyl-CoA to produce acetoacetyl-CoA and drives the reaction to PHA production.
  • the generation of carbon dioxide from the condensation reaction efficiently pushes the reaction towards the production of acetoacetyl CoA (Okamura E et al. (2010) Proc Natl Acad Sci USA 107: 11265-11270).
  • USM2 (phaC Cs ) (Bhubalan K et al.
  • the vector plasmid pTKP2031V was used to introduce a heterologous gene via homologous recombination between the sites of slr2030 and slr2031 (Satoh S et al. (2001) J Biol Chem 276 : 4293-4297).
  • Synechocystis was transformed with a plasmid carrying the phaC Cs gene, nphT7 Ss gene, and C. nekatol acetoacetyl CoA reductase (phaB Cn ) gene under the control of the light-inducible psbAII promoter.
  • C Cs NphT7B Cn A successful transformant strain, C Cs NphT7B Cn , was analyzed for PHA production under a two-stage culture system consisting of a continuous cell growth stage and a PHA accumulation stage.
  • the C Cs NphT7B Cn strain was facilitated by direct photosynthesis of PHA from CO 2 and reached a maximum 14 wt% P (3HB) capacity on day 7 of culture (FIG. 1A).
  • C Cs A Cn B Cn in P under the same culture conditions (3HB) capacity to express phaB Cn (7wt%) reduction observed in It was.
  • RNA-seq library was prepared from cells cultured for 7 days in nitrogen-deficient BG-11 under photoautotrophic conditions. Sequencing was performed using an Illumina platform that yielded a total of 93 million reads (average of 15.5 million reads per sample) for 6 samples. Scatter plots between two biological replicates for each recombinant feline city sample showed a correlation coefficient between 0.96-0.98, indicating the reproducibility of the sequence data.
  • the expression level of each gene was quantified as RPKM (reads per kilobase of exon model per million mapped reads).
  • RNA-seq data provides detailed information on genes that are regulated in response to photoautotrophic PHA accumulation conditions in recombinant Synechocystis strains pTKP2013V, C Cs A Cn B Cn, and C Cs NphT7B Cn .
  • genes that showed higher expression in Synechocystis were primarily involved in photosynthesis, electron transport chains, protein metabolic processes, and nucleic acid metabolism.
  • genes involved in porphyrin and chlorophyll metabolism Two genes involved in porphyrin and chlorophyll metabolism, magnesium-protoporphyrin IX monomethyl ester cyclase (sll1874) and protoheme IX fanesyltransferase (sll1899), were up-regulated in both strains.
  • genes encoding proteins involved in several aspects, such as photosynthetic activity, such as metabolism of photosystems I and II, cytochromes, and chlorophyll are up-regulated in recombinant Synechocystis that actively synthesizes PHA. It was.
  • the transcription level of the gene encoding the protein involved in protein metabolism was decreased in the recombinant Synechocystis strains C Cs A Cn B Cn and C Cs NphT7B Cn .
  • Genes encoding these proteins [DNA mismatch protein (MutL) methionine sulfoxide reductase B (sll1680), prohibitin (slr1106), exozyme S synthetic protein B (ExsB), 3-dehydroquinate dehydrator (AroQ) and hydrogenase (HypA)]
  • This decrease in transcription levels may be related to decreased growth of Synechocystis under nitrogen deficient conditions.
  • Cells respond to nutrient-restricted conditions by a decrease in metabolic activity that occurs concomitant with PHA accumulation.
  • Genes involved in cofactor and vitamin metabolism [lipopeptide antibiotics iturin a biosynthetic protein (slr0495), cobalamin synthase (CobS), 4-hydroxythreonine-4-phosphate dehydrogenase (PdxA), cobalt precorin Decreased expression levels of 6x reductase (CobK), riboflavin biosynthetic protein (RibG), repolitol transferase (LipB), and o-succinylbenzoate synthase (sll0409)] were observed.
  • proteins involved in several aspects of photosynthetic activity such as uroporfinogen decarboxylase (HemE), ferredoxin component (slr1205), protochlorophyllidoductase subunit (BchB), protohome IX fanesyltransferase (CtaB ), Genes encoding photosystem II reaction center protein N (PsbN), and iron stress chlorophyll II binding protein (IsiA) are up-regulated in C Cs NphT7B Cn strain compared to C Cs A Cn B Cn strain It was.
  • HemE uroporfinogen decarboxylase
  • ferredoxin component slr1205
  • BchB protochlorophyllidoductase subunit
  • CtaB protohome IX fanesyltransferase
  • PsbN photosystem II reaction center protein N
  • IsiA iron stress chlorophyll II binding protein
  • NtcA is known to regulate the expression of many genes involved in nitrogen metabolism (Bradley RL, Reddy KJ (1997) J Bacteriol 179: 4407-4410), and the induction of the gene encoding this protein is It may be related to nitrogen-deficient culture conditions applied to increase PHA synthesis in Synechocystis. Conversely, down-regulation of genes involved in DNA binding, transport, translation, and DNA repair was observed in the C Cs NphT7B Cn strain.
  • PHA which is one of the bioplastics
  • the present invention can greatly contribute to the production of various chemicals and biofuels using PHA.

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Abstract

Grâce à de l'introduction de gènes codant l'acétoacétyl-CoA synthase dans des cyanobactéries, le procédé de la présente invention permet de produire du PHA avec un niveau de rendement significativement élevé, même lorsque la culture est réalisée dans une solution de culture à base de sels inorganiques, exempte de sucre, et que la photosynthèse est effectuée par utilisation de CO2 dans l'air en tant que source de carbone.
PCT/JP2014/082073 2013-12-04 2014-12-04 Procédé de production de polyhydroalcanoate utilisant uniquement la photosynthèse Ceased WO2015083769A1 (fr)

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JP2020536520A (ja) * 2017-10-04 2020-12-17 ランザテク，インコーポレイテッド Ｗｏｏｄ−ｌｊｕｎｇｄａｈｌ微生物におけるポリヒドロキシブチレートの生成

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JP2020536520A (ja) * 2017-10-04 2020-12-17 ランザテク，インコーポレイテッド Ｗｏｏｄ−ｌｊｕｎｇｄａｈｌ微生物におけるポリヒドロキシブチレートの生成
JP7399082B2 (ja) 2017-10-04 2023-12-15 ランザテク，インコーポレイテッド Ｗｏｏｄ－ｌｊｕｎｇｄａｈｌ微生物におけるポリヒドロキシブチレートの生成

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