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WO2009140929A1 - Procédé de co-production de 1,3-propanediol, 2,3-butanediol et d'acide polyhydroxypropionique par fermentation de bactéries produites par génie génétique - Google Patents

Procédé de co-production de 1,3-propanediol, 2,3-butanediol et d'acide polyhydroxypropionique par fermentation de bactéries produites par génie génétique Download PDF

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WO2009140929A1
WO2009140929A1 PCT/CN2009/072615 CN2009072615W WO2009140929A1 WO 2009140929 A1 WO2009140929 A1 WO 2009140929A1 CN 2009072615 W CN2009072615 W CN 2009072615W WO 2009140929 A1 WO2009140929 A1 WO 2009140929A1
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pdo
fermentation
php
bdo
genetically engineered
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PCT/CN2009/072615
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Chinese (zh)
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刘德华
刘宏娟
欧先金
雷跃勇
刘卫斌
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清华大学
湖南海纳百川生物工程有限公司
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    • 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
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/18—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic polyhydric
    • 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
    • C12N1/32—Processes using, or culture media containing, lower alkanols, i.e. C1 to C6
    • 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
    • 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.)
    • C12N9/0008—Oxidoreductases (1.) acting on the aldehyde or oxo group of donors (1.2)
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/1025—Acyltransferases (2.3)
    • C12N9/1029—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)
    • 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
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/42—Hydroxy-carboxylic acids
    • 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
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

Definitions

  • the invention belongs to the field of biochemical technology, and particularly provides a method for constructing genetic engineering bacteria fermentation and co-production of PDO, BDO and PHP; the PDO is abbreviated as 1, 3-propanediol, and BDO is abbreviated as 2,3-butanediol.
  • PHP is a polymer (poly ⁇ -hydroxypropionic acid) produced by polymerization of a monomer which is ⁇ -hydroxypropionic acid.
  • 1,3-propanediol is an important chemical raw material that can be used as an organic solvent in inks, printing and dyeing, coatings, lubricants, antifreeze and other industries.
  • the main use of PDO is as a monomer for the synthesis of polyester and polyurethane, especially polybutylene terephthalate ( ⁇ ) formed by polymerization with terephthalic acid, showing a ratio of 1,2-propanediol, dibutyl Alcohol and ethylene glycol are more excellent properties of the polymer synthesized by the monomer.
  • PTT polyethylene terephthalate
  • PTT fiber has the advantages of abrasion resistance, low water absorption, low static electricity, etc., and can compete with nylon in the field of carpet. It can also be used for non-woven fabrics, engineering plastics, garments, home decoration, linings, fabrics, etc. with excellent properties.
  • PTT was named one of the six major petrochemical products in the United States in 1998 and is considered to be an upgraded product of PET.
  • Biosynthesis of PDO is produced by microbial disproportionation of glycerol.
  • glycerol can be converted to PDO microorganisms are mainly anaerobic or facultative anaerobic bacteria, of which Klebsiella pneumoniae, Clostridium butyric and Citwbacter freundii have higher PDO conversion rate, and high tolerance to glycerin and product PDO, therefore has a high development value and application prospects.
  • BDO 2,3-butanediol
  • BDO is a by-product of the production of PDO by fermentation and is also an important chemical raw material. It is a colorless, odorless liquid that can be used as a fuel to prepare polymers, inks, perfumes, antifreeze agents, fumigants, moisturizers, softeners, plasticizers, explosives, and chiral carriers for pharmaceuticals. BDO can also be used as a valuable chemical raw material to synthesize other chemicals. For example, BDO dehydration can produce methyl ethyl ketone. The application of methyl ethyl ketone is quite extensive, and further dehydration can form 1, 3-butadiene.
  • BD0 can be polymerized by Diels-Alder reaction to form styrene.
  • BD0 is condensed with methyl ethyl ketone and hydrogenated to form octane, which can be used to produce high quality flying materials.
  • BD0 reacts with acetic acid to form 2,3-butanediol diacetate, which can be added to the cream to improve flavor.
  • acetic acid to form 2,3-butanediol diacetate, which can be added to the cream to improve flavor.
  • due to its low yield in PDO fermentation it is generally not isolated and purified as a product.
  • Poly- ⁇ -hydroxy fatty acid English name Polyhydroxyalkanoates, abbreviated as PHAs, generally refers to a biopolymer material obtained by dehydration and esterification of hydroxyl and carboxyl groups between monomeric ⁇ -hydroxy fatty acids. It has been found that ruthenium polyester has at least 125 different monomer structures, and new monomers are constantly being discovered.
  • the ruthenium synthesized by microbes has some special properties including biodegradability, biocompatibility, piezoelectricity and optical activity.
  • ⁇ Can meet the needs of a variety of human tissues and organs, such as: cardiovascular system, corneal pancreas, gastrointestinal system, kidney, genitourinary system, musculoskeletal system, nervous system, teeth and mouth, skin and so on.
  • the products that have been commercialized mainly include PHB, PHBV and ⁇ ⁇ .
  • the properties of the crucible can vary from hard to soft to elastic. There are many potential application prospects, and a large number of basic and application development studies are carried out at home and abroad.
  • the simplest monomer of PHAs is ⁇ -hydroxypropionic acid, and the polymer produced by its polymerization is referred to as PHP (polyhydroxypropionic acid).
  • PHP polyhydroxypropionic acid
  • the direct monomer for PHA biosynthesis is ⁇ -hydroxyacyl-Coenzyme, a monomer of this type containing high-energy chemical bonds.
  • sputum can be produced by glycerol metabolism intermediates (see Figure 1).
  • Zeng et al [Sun JB., Heuvel J., Soucaille P., Qu Y., and Zeng AP Comparative Genomic Analysis of dha Regulon and Related Genes for Anaerobic Glycerol Metabolism in Bacteria. Biotechnol. Prog. 2003 19:263-272]
  • the plasmid containing the gene encoding glycerol dehydratase and PDO oxidoreductase was inserted into the wild strain, and the activity of the two enzymes was greatly improved. However, in the actual fermentation process, the engineered bacteria did not Produces high concentrations of PDO.
  • pneumoniae M5al Genetically engineered bacteria with loss of acetic acid, ethanol and lactic acid metabolic pathways, glycerol conversion rate increased, but the final concentration of PDO and production intensity decreased. [Yang Guang. 1, 3-propanediol-producing bacteria M. pneumoniae molecular breeding [ D]. Beijing: China Agricultural University, 2003].
  • Huang Zhihua et al [Huang Zhihua, Zhang Yanping, Cao Zhuan et al. Expression and functional analysis of formate dehydrogenase in Klebsiella pneumoniae. Acta Microbiologica Sinica, 2007, 47 C1): 64 ⁇ 68] obtained from the C. boidinii genome with reduced form
  • the formate I (NADH 2 ) regenerating function of the formate dehydrogenase gene, the formic acid dehydrogenase recombinant plasmid was constructed, and the NADH 2 regeneration system was constructed for the first time in the PDO production strain K. pneumoniae.
  • the object of the present invention is to provide a method for constructing genetic engineering bacteria fermentation to jointly produce PDO, BDO and PHP.
  • PDO producing bacteria knock out D-type lactate dehydrogenase, introduce coenzyme A-dependent aldehyde dehydrogenase and poly
  • the hydroxy fatty acid synthase gene increases the synthesis of NADH 2 in cells, reduces the yield of unproductive by-product lactic acid, and increases the yield of beneficial by-product BDO.
  • the method for constructing genetically engineered bacteria fermentation combined with fermentation to produce PDO, BDO and PHP (as shown in Fig. 2) used in the present invention comprises the steps of: knocking out the D-type lactate dehydrogenase gene in a wild strain producing PDO, introducing Coenzyme A-dependent aldehyde dehydrogenase and polyhydroxy fatty acid synthase gene, constructing genetically engineered bacteria for fermentation combined production of PDO, BDO and PHP; adopting aerobic fermentation and mixing and adding glycerol and alkali solution to the fermentation regulation mode; And the fermentation process is carried out by membrane filtration, electrodialysis, concentration, rectification and the like to separate the product extraction process of the product PDO and BDO and PHP.
  • the specific contents are as follows:
  • step B Using the genomic sample purified in step A as a template, primers were designed based on the D-type lactate dehydrogenase gene, and PCR (polymerase chain reaction) amplification experiments were carried out.
  • the PCR product D-type lactate dehydrogenase gene fragment is purified and ligated to a cloning vector (eg, pMD18-T-vector).
  • a cloning vector eg, pMD18-T-vector
  • PHA synthase (PhaC) gene fragment derived from Ralstonia or Cupriavidus was ligated with PAD_pDK6 to construct the recombinant vector PAD_PhaC_pDK6 (Fig. 3).
  • the PAD_PhaC_pDK6 recombinant vector was transformed into the competent cells of the engineered bacteria in which the D-type lactate dehydrogenase gene was knocked out in the step (1), and the positive clone was identified and isolated, which was the target strain.
  • the constructed genetically engineered bacteria were cultured on a solid medium for 16 to 24 hours, and then placed in a seed culture medium at 30 to 37 ° C for aerobic culture, and the fermentation amount of glycerol was used as a fermentation substrate at an inoculation amount of 1% to 5%. Medium. Fermentation temperature 30 ⁇ 37 °C.
  • the glycerol and alkali solution (1:0.05 ⁇ 1.0) were coupled and fed to control the fermentation. The pH value was controlled at 5.0 ⁇ 8.0, and the flow was stopped after 40 ⁇ 60h. Until the end of the fermentation.
  • the fermentation broth is subjected to microfiltration and ultrafiltration.
  • the filtrate is desalted by electrodialysis. After desalting, it is rich in PDO.
  • the light chamber liquid of BDO is separated by concentration and rectification to separate the product PDO and BDO.
  • the resulting cells were filtered for extraction of PHP.
  • the salt-rich concentrated chamber liquid from electrodialysis is concentrated and crystallized to obtain an organic acid salt such as sodium succinate, which can also be used as one of the products, and the crystallization mother liquid returns to the pre-electrodialysis.
  • the wild strains for constructing genetically engineered bacteria according to the present invention include strains capable of producing PDO, such as Klebsiella, Clostridium citrate, Enterobacter, and Serratia.
  • the genetically engineered fermentation substrate constructed is glycerin, glycerol fermentation broth, biodiesel by-product crude glycerin or by-product crude in the soap industry.
  • step 2 the fermentation is carried out by aerobic fermentation, and a method of mixing and adding fermentation of glycerin and an alkali solution is employed.
  • the genetically engineered bacteria can be simultaneously fermented to produce PDO, BDO and PHP.
  • the fermentation process D-type milk
  • the acid is significantly reduced or not produced, and the post-product extraction process is thus simplified, reducing the cost of energy consumption and post-extraction;
  • BDO and PHP are extracted as products, which increases the added value of the product and improves the utilization of raw materials. Reduced production costs;
  • the introduction of PHP significantly increased the NADH 2 content of the bacteria, which is conducive to the synthesis of PDO.
  • the invention can also be used in the co-production of PDO, BDO and PHP.
  • Figure 1 shows the glycerol metabolic pathway of PHP in microbial cells.
  • HOCH2CH2CH20H is glycerol
  • HOCH2CH2CHO is 3-hydroxypropionaldehyde
  • HOCH2CH2CO-CoA is 3-hydroxypropionyl-CoA
  • Poly(3-Hydroxy-Propionic acid) is poly-3-hydroxypropionic acid
  • CoA is coenzyme A
  • NAD/ NADH is coenzyme I.
  • Figure 2 shows the glycerol fermentation co-production of PHP, PDO and BDO.
  • Glycerol is glycerol
  • 3-Hy droxypropionaldehy de is 3-light propionaldehyde
  • 3-Hydroxy-propionyl-Co A is 3-light-propionyl-CoA
  • Poly(3-Hydroxy-propionic acid) is poly 3-hydroxyl Propionic acid
  • 1,3 -Propanediol is 1,3-propanediol
  • Pyruvate is pyruvic acid
  • Lactate is lactic acid
  • Acetolactate is acetolactate
  • Acetoin is 3-hydroxybutanone
  • 2,3-Butanediol is 2,3-butanediol.
  • Figure 3 shows the recombinant plasmid PAD_PhaC_pDK6.
  • Figure 4 shows the flow chart of PDO, BDO, and PHP product extraction. detailed description
  • the method for synthesizing genetically engineered bacteria to jointly produce PDO, BDO and poly- ⁇ -hydroxypropionic acid comprises knocking out a D-type lactate dehydrogenase gene and introducing a coenzyme-dependent aldehyde dehydrogenase in a wild strain producing PDO And polyhydroxyalkanoate synthase gene, constructing genetically engineered bacteria for producing PDO, BDO and PHP by fermentation; fermenting by fermentation regulation method of mixing and feeding glycerin and alkali solution; and filtering the fermentation broth through membrane, electrodialysis , concentration, distillation and other steps to separate the product extraction process of product PDO and BDO and PHP.
  • Genomic DNA was extracted from Klebsiella HR526 as a starting strain, and a pair of primers GGAATTCACGGTTGCGAACGGTATGTA and GCTCTAGAAGTGGTCTCCGAAATGCTGA was subjected to PCR amplification experiments, and the 5' ends of the primers contained EcoRI and Xbal endonuclease sites, respectively.
  • the PCR product D-type lactate dehydrogenase gene fragment was purified and sequenced. The results were as follows:
  • the above gene fragment was ligated to the cloning vector pMD 18-T-vector.
  • the positive clone vector LDH-pMD18-T-tector was screened, and LDH-pMD18-T-tector was double-digested with EcoRI and Xbal.
  • the recovered fragment was ligated with pGPKm vector which had been digested with the same endonuclease (EcoRI and Xbal). It was then transformed into E. coli SM10 competent cells by electroporation.
  • the SM10 strain containing the recombinant plasmid LDH-pGPKm was subjected to a parental exchange experiment with the wild-type strain, and the strain having the D-type lactate dehydrogenase gene deletion was selected using the kanamycin resistance gene on the pGPKm vector as a selection marker.
  • PAD CoA-Dependent Proionaldehyde Dehydrogenase
  • Primers were designed. For: 5-GCT GAATTC ATGAATACAGCAGAACTGGA-3 and 5-GGCGGTACC TTAGCGAATGGAAAAACCGT-3).
  • the recombinant expression vector PAD_pDK6 was constructed by ligating the vector pDK6 with the help of EcoRI and Kpnl endonuclease.
  • the polyhydroxy acid fatty acid synthase gene fragment derived from Ralstonia eutropha digested with Xba I and B Hind III was ligated with the vector PAD_pDK6 to construct a recombinant vector PAD_PhaC_pDK6.
  • the electroporation method was transformed into the competent cells of the engineered bacteria which were knocked out in the step (1) and the D-type lactate dehydrogenase gene was knocked out, and the positive clone was identified and isolated, which was the target strain HR526G.
  • Fermentation method The genetically engineered bacteria were cultured on a solid medium for 24 hours, and the strain was inserted into a seed medium containing 30 g/L glycerol (250 ml flask, liquid volume 100 ml), culture temperature 37 °C, shaker speed 150rpm, aerobic culture for 24h. A fermented medium containing 30 g/l of initial glycerol was introduced at a 5% inoculation amount. The fermentation was carried out using a 5 L fermenter at a fermentation temperature of 37 °C.
  • the glycerin and NaOH solution (1:0.1) were coupled and flowed, the pH was controlled at 6.0, the aeration was 0.2wm air, and the rotation speed was 150rpm. After 40h, the flow was stopped and the fermentation was completed.
  • OD 65()nm reached 11; PHP content was 1.46% (g/g cell); PDO: 42g/l; BDO: 14g/l, lactic acid was not detected.
  • the fermentation broth is filtered through a membrane, and the filtrate is separated by electrodialysis, concentration, distillation, distillation, and the like to separate the product PDO and BDO.
  • the resulting cells were filtered for extraction of PHP.
  • Product PDO extraction yield 85 %.

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Abstract

Procédé de co-production de 1,3-propanediol (PDO), 2,3-butanediol (BDO) et d'acide polyhydroxypropionique (PHP) par l'intermédiaire d'une fermentation par des bactéries produites par génie génétique. Ce procédé consiste à: désactiver par knock-out la lactate déshydrogénase de type D provenant de bactéries productrices de PDO; introduire des gènes d'aldéhyde déshydrogénase dépendante du coenzyme A et de synthase d'acides gras polyhydroxylés; construire des bactéries par génie génétique pour produire PDO, BDO et PHP; effectuer une fermentation aérobie et régler la fermentation par apport d'un mélange coulant de glycérine et d'une solution basique; filtrer le bouillon de fermentation au moyen d'une membrane et le traiter par électrodialyse, concentration et rectification séparant PDO, BDO et PHP. Les bactéries obtenues par génie génétique peuvent produire simultanément PDO, BDO et PHP, ce qui augmente le rapport d'utilisation des matières premières, limite les coûts, diminue la génération de produits secondaires à base de lactate, simplifie la technique de post-extraction, abaisse les coûts d'extraction et augmente la génération de nicotinamide adénine dinucléotide 2 (NADH2) pendant l'introduction de PHP.
PCT/CN2009/072615 2008-05-04 2009-07-03 Procédé de co-production de 1,3-propanediol, 2,3-butanediol et d'acide polyhydroxypropionique par fermentation de bactéries produites par génie génétique WO2009140929A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102154387A (zh) * 2010-12-27 2011-08-17 山东大学 利用生物柴油副产物生产琥珀酸和聚羟基脂肪酸酯的方法
CN114958928A (zh) * 2022-02-28 2022-08-30 江苏大学 一种基因工程菌发酵联产3-羟基丙酸和1,3-丙二醇的方法

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101307336B (zh) * 2008-05-04 2011-08-17 清华大学 构建基因工程菌发酵联产pdo、bdo和php的方法
US8129169B2 (en) * 2009-06-04 2012-03-06 Genomatica, Inc. Microorganisms for the production of 1,4-butanediol and related methods
US8227217B2 (en) 2009-11-17 2012-07-24 Tsinghua University Methods and genetically engineered micro-organisms for the combined production of PDO, BDO and PHP by fermentation
KR20120099315A (ko) * 2011-01-26 2012-09-10 삼성전자주식회사 3-하이드록시프로피온산 및 1,3-프로판디올 동시 생산용 재조합미생물
CN104531786A (zh) * 2014-12-10 2015-04-22 燃点科技（天津）有限公司 一种应用乙酰辅酶a合成聚3-羟基丙酸的方法
CN108522290A (zh) * 2017-03-02 2018-09-14 云南纳博生物科技有限公司 一种自发光烟草及转基因方法
CN118834117A (zh) * 2024-04-29 2024-10-25 苏州苏震生物工程有限公司 一种2,3-丁二醇的分离方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1348007A (zh) * 2001-04-27 2002-05-08 大连理工大学 一种微生物微氧发酵生产1,3－丙二醇的方法
CN1763210A (zh) * 2005-10-20 2006-04-26 清华大学 微生物好氧发酵生产1,3－丙二醇的方法
CN100999742A (zh) * 2007-01-09 2007-07-18 清华大学 一种发酵生产1，3－丙二醇的高产工艺
CN101307336A (zh) * 2008-05-04 2008-11-19 清华大学 构建基因工程菌发酵联产pdo、bdo和php的方法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1348007A (zh) * 2001-04-27 2002-05-08 大连理工大学 一种微生物微氧发酵生产1,3－丙二醇的方法
CN1763210A (zh) * 2005-10-20 2006-04-26 清华大学 微生物好氧发酵生产1,3－丙二醇的方法
CN100999742A (zh) * 2007-01-09 2007-07-18 清华大学 一种发酵生产1，3－丙二醇的高产工艺
CN101307336A (zh) * 2008-05-04 2008-11-19 清华大学 构建基因工程菌发酵联产pdo、bdo和php的方法

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
HAO, J. ET AL.: "Desalination of Fermented Broth Containing 1,3-Propanediol by Electrodialysis.", THE CHINESE JOURNAL OF PROCESS ENGINEERING, vol. 5, no. 1, February 2005 (2005-02-01), pages 36 - 39 *
QI, W.T. ET AL.: "Progress in Metabolic Pathway and Genetic Regulation of Glycerol Bioconversion to 1, 3-propanediol.", JOURNAL OF CHINA BIOTECHNOLOGY., vol. 23, no. 2, February 2003 (2003-02-01), pages 64 - 68 *
YANG, G ET AL.: "Fermentation of 1,3-propanediol by a lactate deficient mutant of Klebsiella oxytoca under microaerobic conditions.", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, vol. 73, no. 5, January 2007 (2007-01-01), pages 1017 - 1024 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102154387A (zh) * 2010-12-27 2011-08-17 山东大学 利用生物柴油副产物生产琥珀酸和聚羟基脂肪酸酯的方法
CN102154387B (zh) * 2010-12-27 2013-01-30 山东大学 利用生物柴油副产物生产琥珀酸和聚羟基脂肪酸酯的方法
CN114958928A (zh) * 2022-02-28 2022-08-30 江苏大学 一种基因工程菌发酵联产3-羟基丙酸和1,3-丙二醇的方法
CN114958928B (zh) * 2022-02-28 2024-05-14 江苏大学 一种基因工程菌发酵联产3-羟基丙酸和1,3-丙二醇的方法

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