Jiangnan University

Taixing Dongsheng Bio-tech Co., Ltd

Jingwen Zhou

Jian Chen

Guocheng Du

Weizhu Zeng

Meng Ning

Hongwei Guo

Fang Fang

Qiuju Zhang

Yongkun Lv

Xiaomei Cao

Jun Fang

Jianghua Li

Li Liu

Song Liu

Jie Chen

Tao Yang

Wei Lu

Bohan Zhang

Fei Yu

Yuan Qian

Xinrui Zhao

Song Gao

Qian Zhang

Xiaolin Liang

Zhaojun Wang

Jurong Ping

Jie Xu

Disclosed is recombinantfor producing L-tyrosine and application thereof, and belongs to the technical fields of genetic engineering and bioengineering. According to the present disclosure, genes aroP and tyrP are knocked out, expresses the endogenous gene yddG of, then heterologously expresses fpk from, expresses the endogenous genes ppsA and tktA of, and then expresses aroGand tyrA. Knocking out tyrR, trpE and pheA, so that the synthesis flux of L-tyrosine is increased. Finally, an endogenous gene poxB is knocked out to realize stable fermentation performance at high glucose concentration.

Recombinantfor producing l-tyrosine and application thereof

The present disclosure provides an automatic production line for manufacturing and processing plant protein meat, and belongs to the field of application of food equipment. The automatic production line for manufacturing and processing the plant protein meat comprises: an extrusion-expansion machine, plant protein meat raw material blocks, a bearing plate, a first manipulator, a storage rack, a first conveying device, a second manipulator, a second conveying device, a main console, a third conveying device, a shredding device, a seasoning adding and mixing device, a third manipulator, a recycling rack, a storage cabinet and a secondary console. The shredding device of the present disclosure shreds the plant protein meat raw material blocks by cooperative up-down staggered motion of double rows of clips, the plant protein meat raw material blocks in any shape can be shredded, and breakage of the raw material shreds in the shredding process can be avoided effectively; and by the seasoning adding and mixing device of the present disclosure, the shredded plant protein meat raw material blocks are added with seasonings and mixed with the seasonings. The present disclosure is simple in structure, facilitates manufacturing of the plant protein meat, and can efficiently produce and process the plant protein meat.

Automatic production line for manufacturing and processing plant protein meat

The disclosure discloses construction of recombinant Saccharomyces cerevisiae for synthesizing carminic acid and application thereof and belongs to the technical field of genetic engineering and bioengineering. The disclosure obtains recombinantCA-B2 capable of synthesizing carminic acid by heterologously expressing cyclase Zhul, aromatase ZhuJ, OKS of Octaketide synthase 1, C-glucosyltransferase UGT2, monooxygenase aptC and 4'-phosphopantetheinyl transferase npgA in. The recombinantcan be used for synthesizing carminic acid by taking self-synthesized acetyl-CoA and malonyl-CoA as a precursor. On this basis, OKS, cyclase, aromatase, C-glucosyltransferase and monooxygenase relevant to carminic acid are integrated to a high copy site, which can remarkably improve the yield of carminic acid. The yield of carminic acid can be increased to 2664.6 &#x3bc;g/L by optimizing fermentation conditions, and the fermentation time is shortened significantly. Therefore, the recombinantplays an important role in the fields of cosmetics, textiles and food.

Construction of recombinantfor synthesizing carminic acid and application 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 disclosure relates to a method for improving the yield and production intensity of() sorbose, and belongs to the technical field of fermentation engineering. By knocking out genes related to formation of D-sorbitol or L-sorbose metabolic by-products in, the formation of the by-products is reduced, and the efficiency of transforming D-sorbitol into L-sorbose is improved, thereby improving the yield and production intensity of L-sorbose. A recombinant strain-11 constructed by the present disclosure, compared with a control strain, has an L-sorbose transformation rate of 96.12%, which is 4.47% higher than that of a wild strain, has a production intensity of 14 g/L&#xb7;h, which is 14.7% higher than that of the wild strain, and has a fructose by-product content of only 5.6 g/L, which is 45.6% lower than that of the wild strain.

Method for improving yield and production intensity ofsorbose

The present disclosure provides a method for extracting alpha-ketoglutarate and pyruvate simultaneously from microbial fermentation broth or enzyme transformation solution, which is related to the technical field of biological separation and extraction. The method comprises the following steps: centrifuging the microbial fermentation broth or enzymatic conversion solution containing &#x3b1;-KG and PA to remove the cells and other visible solids; removing the macromolecular impurities by ultrafiltration; evaporating and concentrating under reduced pressure conditions; extracting with the water-insoluble extraction after acidification; separating crude crystals of &#x3b1;-KG and crude liquid of PA by evaporation crystallization method (if concentration of PA is great higher than that of &#x3b1;-KG, crystallization separation should be conducted after distilling partial pure pyruvate); washing the crude crystal of &#x3b1;-KG with water-insoluble organic solvent as ethyl acetate or butyl acetate, drying and crushing to obtain qualified &#x3b1;-KG; distilling to gain qualified PA product applying high vacuum distillation (or molecular distillation).

Method for extracting alpha-ketoglutarate and pyruvate simultaneously from microbial fermentation broth or enzymatic conversion solution

The invention provides a high-throughput screening system based on multi-manipulators, and belongs to the field of biotechnology and detection equipment. A high-throughput screening system based on multi-manipulators, comprises of the first manipulator, sampler, pipette, plate washer, microplate reader, the second manipulator, centrifuge, deep-well plate library, waste shallow-well plate barrel, shallow-well plate library, waste needle plate barrel, needle library, waste deep-well plate barrel, collection box. The present invention is a combination of microbiology and mechanics. The aim of the invention is to realize the automation and intelligentization of the high throughput screening experiment, effectively improve the experimental accuracy, reliability and efficiency. It contributes to the development of high throughput screening technology for microorganisms and drugs.

High-throughput screening system based on multi-manipulators

The present disclosure provides a multicopper oxidase mutant with improved salt tolerance. Threonine at site 317 of wild-type multicopper oxidase WT was mutated to asparagine, leucine at site 386 was mutated to tyrosine, and serine at site 427 was mutated to glutamic acid by site-directed mutagenesis to obtain a mutant T317N-L386Y-S427E. Compared with WT, the tolerance of T317N-L386Y-S427E to 6%, 9%, 12%, 15% and 18% NaCl (W/V) is improved.

Multicopper oxidase mutant with improved salt tolerance

The present invention provides a production-line-type high-throughput screening system, which relates to the field of biotechnology and testing equipment. The system comprises of four manipulators, three parallel conveyor belts with fixed slots, 2-DOF slipway and fixed fixtures, 96-channel pipetting system, coloring device, oscillating mixing device, microplate reader, well plates loading platform and well plates recycling platform. Manual operation takes five minutes to detect one 96-well plate, while this system can handle 20 96-well plates per minute. It expands the number of screening targets, making the screening process more clearly and concisely and liberating manual labor. The system makes effective contributions to the development of microbial breeding technology.

Production-line-type high-throughput screening system

The present invention provides a method for high-efficiency production of pinoresinol by use of an HOauto-scavenging enzymatic cascade. It uses eugenol as the substrate, which is relatively inexpensive and is industrially available. It uses an enzymatic cascade to remove HOproduced in the process of pinoresinol synthesis, thereby reducing its inhibitory effect on the enzyme activity. In addition, the present invention uses whole cells as a catalyst, which can continuously regenerate cofactors needed by the enzyme, thus eliminating the need for exogenous addition of expensive cofactors during the reaction. The yield of the present invention can reach 7.12 g/L and the conversion rate is 61.55%.

Method for high-efficiency production of pinoresinol using an HOauto-scavenging cascade

The present invention provides a method for decreasing pyruvate catabolism and increasing the accumulation of pyruvate in microbes. By overexpressing wild type dihydrolipoamide acetyltransferase or dihydrolipoamide acetyltransferase mutants which have mutations at conservative active sites, the present invention provide a method to decrease overall activity of pyruvate dehydrogenase complex and pyruvate catabolism, and thus increase the accumulation of extracellular pyruvate without killing the pyruvate-producing microbes. Overexpressing dihydrolipoamide acetyltransferase mutants is an effective way to increase pyruvate accumulation.

Method for decreasing pyruvate catabolism and increasing the accumulation of pyruvate in microbes

The present invention provides a genetically engineeredwith enhanced extracellular secretion of pyruvic acid.strain was obtained from China Center for Type Culture Collection with CCTCC No: M202019 and over-expressed the optimized CutA (SEQ ID NO:2) encoding stress protein. Both of the temperature tolerance ofand extracellular concentration of pyruvate were improved by overexpressing the optimized CutA. The optimum growth temperature of genetically engineeredwas increased too. The present invention can be widely used to increase extracellular levels of pyruvate during the fermentation process.

[object Object]

The present invention provides four keto acid transporter encoding sequences selected from 6611 protein coding sequences ofCLIB122 database. Also provided are recombinantstrains overexpressing the keto acid transporters, which have increased extracellular secretion of &#x3b1;-ketoglutarate. The present invention can be used to increase extracellular levels of &#x3b1;-ketoglutarate during the fermentation process and lower downstream purification cost for &#x3b1;-ketoglutarate production.

Growing community of inventors

Wuxi, China

Jingwen Zhou