Jiangnan University

Shandong Runde Biotechnology Co., Ltd

Bright Dairy & Food Co., Ltd.

Bloomage Biotechnology Corporation Limited

Taixing Dongsheng Bio-tech Co., Ltd

Inner Mongolia Mengniu Dairy (group) Co., Ltd.

Jianghua Li

Jian Chen

Guocheng Du

Long Liu

Xueqin Lv

Jieying Deng

Xian Yin

Yanfeng Liu

Jianxing Lu

Changfeng Liu

Yang Gu

Cheng Zhang

Xiaodong Jiang

Zhijie Hu

Sai Jin

Fuxin Sun

Jianwei Jiang

Taichi Chen

Yaokang Wu

Tengfei Niu

Wei Lu

Chen Deng

Jiangong Lu

Yang Wayne Wang

Jingwen Zhou

Zhen Kang

Juan Zhang

Chunmei Chen

Xinrui Zhao

Chenyang Zhang

Xianhao Xu

Miao Wang

Xiaomin Dong

Litao Hu

Jialian Li

Tianmeng Zhang

Guolin Zhou

Wenyang Wu

Yuan Qian

Fei Yu

Bohan Zhang

Li Sun

Xiaolong Zhang

Xueliang Qiu

Zhuolin Feng

Ningzi Guan

Yawen Zhao

Yang Song

Hannes Link

Hongzhi Zhang

Zhu Jiang

Ke Liu

The invention provides astrain for producing a human milk lipid substitute. By integrating a heterologous lysophosphatidic acid acyltransferase intoand knocking out its own natural lysophosphatidic acid acyltransferase, the content of palmitic acid (C16:0) at Sn-2 position of triacylglycerol produced byis increased, to synthesize a human milk lipid substitute. On this basis, a metabolic pathway related gene is knocked out, to further increase the content of human milk lipid substitute in the product. In the present invention, a human milk lipid substitute is de novo synthesized byfor the first time, in which the total fatty acid is 15% or more, and the relative content of C16:0 at Sn-2 position reaches about 60%.

strain for producing human milk lipid substitute and use thereof

The present invention discloses a RamA transcription factor mutant for promoting the production of N-acetylglucosamine and use thereof. The mutant is obtained by mutating lysine at position 90 to asparagine and serine at position 92 to lysine in a parent having an amino acid sequence as shown in SEQ ID NO: 2. The present invention provides a genetically engineered strain that overexpresses the RamA transcription factor mutant and increases the production of N-acetylglucosamine. By overexpressing the transcription factor RamA that is involved in the regulation of carbon metabolism, the extracellular accumulation of N-acetylglucosamine is increased, with a maximum concentration reaching 31.5 g/L, which lays a foundation for further metabolic engineering ofto produce glucosamine. The method for constructing recombinantof the invention is simple, and convenient in use, and thus has good application prospects.

RamA transcription factor mutant for promoting production of N-acetylglucosamine and use thereof

The present invention provides a method for promoting N-acetylglucosamine synthesis by using the GlcN6P responsive element. In the present invention,BSGNY-P-glmS-P-GNA1 is used as a starting strain, in which a CRISPRi system regulated by GlcN6P responsive element is integrated into the genome to dynamically weaken the N-acetylglucosamine synthesis competitive pathway; a GlcN6P responsive promoter is used to regulate the expression of GNA1 on the plasmid to dynamically regulate the N-acetylglucosamine synthesis pathway; and the key gene alsSD involved in the acetoin synthesis pathway is knocked out. During fed-batch fermentation with this strain in a 15 L fermenter, the production of N-acetylglucosamine reaches 131.6 g/L and no by-product acetoin is accumulated, which lays a foundation for the production of GlcNAc by industrial fermentation.

Method for promoting n-acetylglucosamine synthesis by using GlcN6P responsive element

The invention discloses a recombinantfor efficient synthesis of highly pure hyaluronic acid and oligosaccharides thereof, belonging to the technical field of bioengineering. The recombinantconstructed in the present invention can produce hyaluronic acid with a yield up to 40 g/L, and a crude product purity of 95%. Addition of exogenous hyaluronic acid hydrolase and optimization of the fermentation conditions results in hyaluronic acid oligosaccharides with specific molecular weight, and can further improve the yield of hyaluronic acid to 72 g/L. The invention lays a solid foundation for the efficient synthesis of highly pure hyaluronic acid by microorganisms, and the constructed recombinantis suitable for industrial production and application.

High-efficiency synthesis and high-purity hyaluronic acid, and recombinantfor oligosaccharide thereof

The disclosure discloses recombinantfor synthesizing e lacto-N-neotetraose yield. The recombinantis obtained by integrating two &#x3b2;-1,4-galactotransferase genes on a genome of a host bacterium168&#x394;amyE:P-lacY, P-lgtB, P-comK and exogenously expressing a &#x3b2;-1,3-N-glucosaminotransferase gene. Compared with a strain before transformation, the recombinantof the disclosure improves the yield of the synthesized lacto-N-neotetraose from 720 mg/L to 1300 mg/L, laying a foundation for further metabolic engineering transformation offor producing the lacto-N-neotetraose.

Recombinantfor synthesizing lacto-N-neotetraose and application thereof

The present invention provides a pyruvate-responsive biosensor and a construction method and use thereof. In the present invention, pyruvate-responsive biosensors with different dynamic ranges are successfully constructed by optimizing the PdhR binding sequence inserted on the P43 promoter and optimizing the insertion site, wherein the minimum increase in dynamic range is by 0.6 time, and the maximum increase is by 30.7 times. The pyruvate-responsive biosensors are useful in the precise control of the expression of each gene in the cell. Since pyruvate is a key metabolite of central carbon in the cells, these biosensors are capable of dynamically regulating the expression level of intracellular genes according to changes in the content of pyruvate in the cells, thereby achieving the dynamic control of intracellular metabolic flux. The pyruvate biosensor obtained in the present invention has a good specificity, and a response range to pyruvate of 10-35 nmol/g DCW.

Pyruvate-responsive biosensor, and construction method and use thereof

The disclosure discloses a genetically engineered strain for producing porcine myoglobin and fermentation and purification thereof, and belongs to the technical field of genetic engineering. The disclosure realizes efficient secretion and expression of porcine myoglobin by integrating the gene of porcine myoglobin in. On this basis, optimization of the medium and culture conditions of recombinantcan significantly increase the titer of porcine myoglobin, so that the titer can reach 285.42 mg/L under fermenter conditions. In addition, by creatively adding different concentrations of ammonium sulfate to fermentation broth step by step, the purity of myoglobin obtained by final concentration is up to 88.0%, and the purification rate is up to 66.1%. The disclosure realizes efficient expression and high purification of porcine myoglobin from various steps such as synthesis, fermentation and purification of porcine myoglobin, and provides broad prospects for industrial production of porcine myoglobin.

Genetically engineered strain for producing porcine myoglobin and food-grade fermentation and purification thereof

The present invention provides an acid-tolerantstrain and use thereof. By using exogenously added malic acid as a stress, an acid-tolerant mutantstrain MTPfo-4 is obtained by directed evolution screening in the laboratory, which tolerates a minimum pH of 2.44. The mutant strain MTPfo-4, tolerant to multiple organic acids, has an increased tolerance to exogenous malic acid of up to 86.6 g/L. The mutant strain MTPfo-4 obtained is further identified. The mutant strain grows stably and well, and can tolerate a variety of organic acids (lactic acid, malic acid, succinic acid, fumaric acid, citric acid, gluconic acid, and tartaric acid). It also has a strong tolerance to inorganic acids (HCl and HPO). This is difficult to achieve in the existing research and reports of. The strain is intended to be used as an acid-tolerant chassis cell factory for producing various short-chain organic acids.

Acid-tolerantand use thereof

The disclosure disclosesfor producing N-acetylneuraminic acid and application thereof, and belongs to the field of genetic engineering. The disclosure optimizes the expression levels of key enzymes in N-acetylneuraminic acid synthesis pathways on genome through promoters of different strength, reduces the protein synthesis pressure caused by the expression of enzymes on cells, and further integrates the three N-acetylneuraminic acids in a sameengineering strain.with improved N-acetylneuraminic acid production is obtained, and the production reaches 10.4 g/L at the shake flask level, laying a foundation for further improving the NeuAc production from

for producing N-acetylneuraminic acid and application thereof

The present disclosure discloses recombinantand application thereof in screening erythritol-producing strains, and belongs to the technical field of microorganisms. The recombinantused in a method for screening an erythritol-producing strain disclosed by the present disclosure can well perform positive correlation induction on erythritol with different concentrations, so that the method for screening the erythritol-producing strain has the advantage of high sensitivity. High-concentration glucose is usually adopted as a fermentation substrate when erythritol is produced in a fermentation mode in the industry, but the method for screening the erythritol-producing strain disclosed by the present disclosure can overcome the interference of the high-concentration glucose, and under the interference of the high-concentration glucose, the recombinantused in the method for screening the erythritol-producing strain can still well perform positive correlation induction on erythritol with different concentrations, and the correlation is higher than that without the interference of the glucose. Therefore, the method for screening the erythritol-producing strain has the advantage of strong anti-interference capability.

Recombinantand application thereof in screening erythritol-producing strains

The disclosure discloses recombinantfor synthesizing guanosine diphosphate fucose and a construction method and application thereof. The recombinantis obtained by intensively expressing guanylate kinase and nucleotide diphosphokinase genes and expressing exogenous fucokinase and phosphate guanylyltransferase genes in a genome of168. According to the disclosure, a bacterial strain for synthesizing the guanosine diphosphate fucose is obtained by reconstructing the168, with a volume of intracellular accumulation up to 196.15 g/L. According to the disclosure, by intensively expressing the guanylate kinase and nucleotide diphosphokinase genes, and enhancing the supply of intracellular GDP-L-fucose composition cofactors, the synthesis of the guanosine diphosphate fucose is promoted. The construction method for the recombinantof the disclosure is simple and convenient to use, thus having good application prospects.

Recombinantfor synthesizing GDP-L-fucose and application thereof

The present invention provides a recombinantproducing N-acetylglucosamine and use thereof. The recombinantis obtained by overexpressing, in, the transcription factor SugR derived therefrom. The recombinantof the present invention increases the production of acetylglucosamine to up to 26 g/L, and lays a foundation for further metabolic engineering ofto produce glucosamine.

Transcription factor SugR coding gene, and use thereof in production of N-acetylglucosamine

The present invention provides a recombinantwith improved 2'-fucosyllactose production, and a construction method thereof. In the present invention, a strain capable of efficiently synthesizing 2&#x2032;-fucosyllactose is obtained by the fusion expression of the fucosyltransferase gene and the L-fucokinase/guanosine 5&#x2032;-diphosphate-L-fucose pyrophosphorylase gene inBSGL-FF, the fermentation supernatant of which comprises a cumulative amount of 2&#x2032;-fucosyllactose as high as 1.62 g/L, which is 55% higher than the amount achieved with the control strain. The construction method of the recombinantof the present invention is simple, and convenient to use, and thus has good application prospects.

Recombinantwith improved 2'-fucosyllactose production, and construction method thereof

The present invention relates to a method for promoting acetylglucosamine synthesis of, which belongs to the field of genetic engineering. The present invention adopts the recombinantBSGNKAP2 as a starting strain, exogenously introducing pyruvate carboxylase BalpycA derived from, eliminating the central carbon metabolism overflow of theand avoiding the synthesis of the by-product acetoin; further, five exogenous reducing force metabolic reactions are introduced to replace the reaction of generating NADH in glycolysis pathway and tricarboxylic acid cycle to reconstruct intracellular reducing force metabolism, which specifically comprise glyceraldehyde-3-phosphate ferredoxin dehydrogenase, isocitrate NADdehydrogenase, a malate quinone dehydrogenase, a ketoacid ferredoxin oxidoreductase and a nitrogenase ferritin. In a shake-flask fermentation process using a complex medium, acetylglucosamine yield of the recombinant strain BSGNKAP8 is 24.50 g/L, acetylglucosamine/glucose yield is 0.469 g/g, respectively 1.97 times and 2.13 times of those of the starting strain BSGNKAP2.

Method for promoting acetylglucosamine synthesis of bacillus subtilis

The invention provides a recombinant, construction method and use thereof, wherein the cell's own FMMs are used as a space scaffold, and a multi-enzyme complex is constructed from specific marker proteins FloA and FloT, such that an artificial substrate channel is formed, and the cell metabolic burden is effectively reduced. The recombinantof the invention can efficiently synthesize GlcNAc without affecting cell life activity, and can also limit the toxic intermediate metabolite GlcN-6-P near the plasma membrane to reduce or eliminate its inhibition on cell activity. In the process of shaking flask fermentation using complex medium, the yield of acetyl glucosamine of the control strain BSG-C was only 0.45 g&#xb7;L, while that of BSG-AT, BSG-ATP, BSG-ATPB increased to 5.29 g&#xb7;L, 6.22 g&#xb7;Land 8.48 g&#xb7;Lrespectively. The construction method of recombinantis simple, easy to use and has a good application prospect.

Recombinantand use thereof

Growing community of inventors

Wuxi, China

Jianghua Li