Jiangnan University

Rutgers, the State University of New Jersey

Yan Xu

Xiaowei Yu

Qun Wu

Yao Nie

Rongzhen Zhang

Gaetano T Montelione

Rong Xiao

Dong Wang

Songyin Zhao

Hai Du

Kunpeng Li

Hongxia Zhang

Nan Deng

Cong Ren

Huilin Wang

Yaohui Li

Jiaojiao Guan

By mutating arginine at position 97 and asparagine at position 43 of a wild type L-proline-3-hydroxylase, a mutated L-proline-3-hydroxylase is obtained. The mutated L-proline-3-hydroxylase comprises a variant of the wild-type L-proline-3-hydroxylase having SEQ ID NO:1, which comprises a substitution of the arginine corresponding to position 97 (R97) of SEQ ID NO:1 and a substitution of the asparagine corresponding to position 43 (N43) of SEQ ID NO:1; wherein the amino acid substituted into the mutated L-proline-3-hydroxylase at position R97 is methionine, and wherein the amino acid substituted into the mutated L-proline-3-hydroxylase at position N43 is selected from the group consisting of threonine (T), alanine (A), valine (V), serine(S), aspartic acid (D), and lysine (K).

Dioxygenase mutants and use thereof in synthesizing (2S,3R)-3-hydroxypipecolic acid

The present disclosure discloses aand a multifunctional complex microbial inoculant and use thereof, and belongs to the technical field of bioengineering. Theof the present disclosure has a degrading ability of lactic acid as high as 12.69 g·L, which is 2.04 times that of a type strain. At the same time, the strain can also metabolize ethanol and has an ODof 4.48 after fermentation in a sorghum juice medium at 30° C. and 200 rpm for 3 d. Thecould completely consume 58 g·Lof glucose in the sorghum juice medium after 60 h of fermentation and produce 13.06 g·Lof ethanol. Thedegrades lactic acid and can relieve a lactic acid pressure of a fermentation system and enableto grow and metabolize to produce wine. In addition, the strain and the microbial inoculant thereof can inhibit the production of filamentous fungi and geosmin and have important use prospects for maintaining homeostasis of a fermentation system and food preservation.

and multifunctional complex microbial inoculant and use thereof

The disclosure disclosessp. LBM10036 capable of utilizing multiple substrates and application thereof, and belongs to the fields of microbial technology and Baijiu brewing. The microorganismsp. LBM10036 of the disclosure was preserved in the Guangdong Microbial Culture Collection Center on Sep. 21, 2020, with the preservation number GDMCC No. 61202. Thesp. LBM10036 of the disclosure has the capability to utilize diverse substrates and high product specificity, adapts to low pH, and can efficiently synthesize caproic acid under a pH of 5.0-5.5. The strain used in the disclosure can be used for strengthening microorganisms in pit mud, and improving the quality of degraded pit mud, to improve the quality of Strong aroma-type Baijiu, and also can be used for producing caproic acid from a variety of sugars and lactic acid by microbial fermentation.

Caproate-producing bacterium with multiple substrate utilization capabilities and its applications

The present invention discloses a method for promotingto synthesize surfactin based on multi-gene synergy, and belongs to the field of genetic engineering. Firstly,is enabled to obtain the ability to synthesize surfactin by integrant expression of sfp protein derived from a high-yield strain, on this basis, by knocking out genes associated with a competitive pathway, overexpressing genes associated with the surfactin tolerance of, strengthening genes associated with a branched chain fatty acid synthesis pathway or improving the intracellular srfA gene transcription level, the synergy among genes is realized, and systemic metabolic engineering transformation is performed on the, thereby greatly improving the ability ofgenetically engineered bacteria to synthesize surfactin. Compared with a starting strain168, the amount of extracellular accumulation of surfactin of thegenetically engineered bacteria increases from 0 g/L to 12.8 g/L. The present invention obtains the recombinantwith significantly improved ability to synthesize surfactin by multi-gene cooperative transformation of, and has a good application prospect.

Method for promotingto synthesize surfactin based on multi-gene synergy

The present invention discloses a low urea-producing and flavor-producingstrain and a use thereof in food production, falling within the fields of wine brewing and food safety. Theof the present invention is obtained by isolating from a liquor fermentation environment (Daqu), is namedCGMCC NO. 12416, and was deposited at China General Microbiological Culture Collection Center on May 6, 2016, with a deposit number of CGMCC NO. 12416. The strain of the present invention has the characteristics of low urea production, flavor production, and tolerance to ethanol and acids, is an excellent strain having a fermentation function, and can be used in brewed wine, distilled liquor and other food fields to ensure food safety.

Low urea-producing and flavor-producingstrain and use thereof in food production

The invention discloses a method for producing surfactin by using, belonging to the technical field of industrial microorganisms. The invention synthesizes surfactin by using the screenedCGMCC No. 12593, the yield is up to 9.43 g/L, and a fermentation process has the characteristics that the surfactin synthesis and the growth of the strain are synchronous without a lag period. The strain and the fermentation method provided by the invention will play an important role in the industrial production of surfactin and have broad application prospects.

Method for producing surfactin by using bacillus amyloliquefaciens

The present disclosure discloses a method for preparing lipase with high ester synthesis activity by using surfactant, belonging to the field of enzyme engineering. The present disclosure provides a method for obtaining a lipase with high ester synthesis activity by adding different surfactants with different concentrations in a lipase aqueous solution and then freeze-drying. The lipase meets the requirement in non-aqueous catalysis. Mixing a variety of lipase with no ester synthesis activity or low activity at suitable concentration and an appropriate concentration of the surfactant in the solution can produces lipases with significantly improved ester synthesis activity, meanwhile changing the hydrolytic activity of the lipase. Increased ester synthesis activity makes lipase more suitable for industrial applications.

Preparation of lipase with improved ester synthesis activity by using surfactants

The invention discloses a high-secretion heat-resistant yeast genetically engineered strain and application thereof, belonging to the field of biotechnology. Mutagenesis and domestication are performed to obtain the yeast genetically engineered strain capable of expressing a lipase gene at a high secretion level at high temperature. The strain is collected by China Center for Type Culture Collection (CCTCC), and the collection number is CCTCC NO: M 2016278. The sequencing analysis shows that the lipase gene and promoter sequence thereof are not mutated, which indicates that the high-secretion expression of the lipase gene by the mutant strain is caused by mutation of other gene sequences in the genome. By knocking out the lipase gene in homologous double-crossover way, the constructed knockout strain can be used as an expression host of other exogenous proteins, can be used for enhancing the expression level of other exogenous genes, and can be subjected to fermentation under high-temperature fermentation conditions.

High-secretion heat-resistant yeast genetically engineered strain and application thereof

Disclosed are methods for the synthesis of chiral alcohols by Sortase A-mediated oxidoreductase oligomers, which relates to the field of biocatalysis. In the present disclosure, oxidoreductase oligomers were used as biocatalysts for chiral alcohol preparation. Compared to wild-type enzymes, the oxidoreductase oligomers significantly improved catalytic activity and thermal stability. The sortase A-mediated oxidoreductase oligomers had 6-8 folds improvement in specific activity over that of the wild-type enzymes. The oligomers displayed a Tvalue 6-12&#xb0; C. higher than that of the wild-type, suggesting the sortase A-mediated oxidoreductase oligomers significantly improved thermostability of the enzymes. The oxidoreductase oligomers catalyzed asymmetric transformation to produce (S)-1-phenyl-1,2-ethanediol or (R)-1-phenethyl alcohol within 3-6 hr, with an optical purity of 98%-100% and a yield of 98%-99%.

Carbonyl reductase oligomers and their application in synthesis of chiral alcohols

The invention discloses a method for improving the extracellular expression level of a foreign protein by means of phospholipase fusion expression. Four proteins, PLA, MBP, CBD and SUMO, are used as a fusion tag to construct a fusion gene. Compared with an original protein MOH without any fusion tag, the extracellular expression level and enzymatic activity of all the four fusion proteins are increased to some degree. Among them, the fusion protein using PLAas the fusion tag has the highest expression level, which is 7.4 times higher than that of the original protein. Compared with other fusion tags, PLAhas a low molecular weight and the fusion protein having PLA2 as the fusion tag has the highest expression level (up to 12 g&#xb7;Lin a 7 L fermentation tank for high-density fermentation). It is shown that the secretory expression of a foreign protein can be effectively increased by using PLAas a fusion tag.

Methods for improving expression levels of foreign proteins by means of phospholipase fusion expression

The present invention provides a novel dual-function lipase mutant and its application in processing of flour products. The amino acid sequence of the lipase has one of the following amino acid substitutions: P298T,P298T/H317P, P298T/H317P/V326S, P298T/T218S/S234F, P298T/H317P/P168L/A129S, P298T/S234F/K161R/V326S, wherein the substitutions are relative to a parent amino acid sequence set forth in SEQ ID NO:1; wherein the lipase mutant maintains a triglyceride and lecithin hydrolysis activity and good performance in thermostability. The mutants have good performance in processing of flour products, while they can significantly whiten the bread or other products in processing of flour products and significantly increase the specific volume in bread baking process.

Bifunctional lipase mutant and methods of using same

The present invention provides a novel dual-function lipase variant and its application in processing of flour products. The amino acid sequence of the lipase has one of the following amino acid substitutions: P298T, P298T/H317P, P298T/H317P/V326S, P298T/T218S/S234F, P298T/H317P/P168L/A129S, P298T/S234F/K161R/V326S or the nucleotide sequence of wherein said lipase is substituted, deleted, or added based on the sequence encoding the amino acid described in (a) and has at least 80% identity with it. The variants have good performance in processing of flour products, while they can significantly whiten the bread or other products in processing of flour products and significantly increase the specific volume in bread baking process.

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Provided are novel hetero-oligomers of (S)-carbonyl reductases and their application in catalyzing the reduction of polyphenyl ketones. Also provided are recombinant strains expressing hetero-oligomers of SCR/SCR2 or SCR2/SCR3, which can catalyze the reduction of polyphenyl ketones. The hetero-oligomer of SCR/SCR2 is capable of catalyzing 2,4-dichlorobenzophenone, 2-naphthalenone and [(E)-2-[3-[3-[2-(7-chloro-2-quinolinyl)ethenyl]phenyl]-3-oxopropyl]benzoic acid methyl ester (Keto Easter M) with a specific activity of 4.55 U/mg, 2.43 U/mg and 0.86 U/mg, respectively. The hetero-oligomer of SCR2/SCR3 is capable of catalyzing 2,4-dichlorobenzophenone and [(E)-2-[3-[3-[2-(7-chloro-2-quinolinyl)ethenyl]phenyl]-3-oxopropyl]benzoic acid methyl ester (Keto Easter M) with a specific activity of 4.42 U/mg and 1.21 U/mg, respectively. No catalyzing activities for reducing polyphenyl ketones were detected in the naturally existing homo-oligomers of SCR, SCR2 or SCR3, which can catalyze the reduction of monobenzoic cyclic compounds. The invention expands the substrate spectrum of natural (S)-carbonyl reductases (SCR, SCR2 and SCR3) and provides a novel type of oxidoreductases for catalyzing the reduction of polyphenyl ketones.

Hetero-oligomers of (S)-carbonyl reductases and their applications in catalyzing reduction of polyphenyl ketones

Stereospecific carbonyl reductases SCR1, SCR2, and SCR3 are described herein as are nucleotide sequences that encode these reductases. These stereospecific carbonyl reductases have anti-Prelog selectivity and have specificities that are useful for fine biochemical synthesis.

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Wuxi, China

Yan Xu