Abstract: The present disclosure provides a recombinant Escherichia coli for producing chlorogenic acid and application thereof. In the present disclosure, tyrosine ammonia-lyase FjTAL derived from Flavobacterium johnsoniae, hpaBC derived from E. coli, 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase mutant aroGfbr, chorismate mutase tyrC derived from Zymomonas mobilis, quinic acid/shikimate-5 dehydrogenase ydiB derived from E. coli, hydroxycinnamoyl CoA:quinic acid transferase NtHQT derived from Nicotiana tabacum, and 4-coumarate:CoA ligase At4CL1 derived from Arabidopsis thaliana are expressed in the recombinant E. coli, thereby constructing a chlorogenic acid biosynthesis pathway in E. coli. Then, the aroB gene and gldA gene derived from E. coli are overexpressed, and an endogenous gene menI is knocked out from the recombinant E. coli. The recombinant strain produced chlorogenic acid by fermentation at a titer of up to 638.2 mg/L in a shake flask or at a titer of 2.8 g/L in a 5-L fermenter.

Abstract: The present disclosure discloses recombinant Escherichia coli for expressing a synthesis pathway of asiaticoside and application thereof, and belongs to the field of bioengineering. Rhamnosyltransferase with an amino acid sequence shown in any one of SEQ ID NO: 25 to SEQ ID NO: 29, glucosyltransferase with an amino acid sequence shown in any one of SEQ ID NO: 10 to SEQ ID NO: 17 and UGT73AH1 reported in a document are transferred into E. coli BL21 (DE3)?pgi to realize co-expression. Fermentation results show that all 5 rhamnosyltransferases screened in the present disclosure can achieve effects, and a unique new peak appears at 0.596 min, which is consistent with a characteristic ion flow of an asiaticoside standard product. According to the present disclosure, barriers of the prior art are broken through, a new biosynthesis method for asiaticoside is provided, and industrialization of biosynthesis of asiaticoside becomes possible.

Abstract: Disclosed is a method for producing phycocyanobilin by use of a recombinant Escherichia coli that express heterologous heme oxygenase ho1 and ferredoxin oxidoreductase pcyA derived from Synechocystis sp. PCC6803. According to the present disclosure, heterologous expression of ho1 and pcyA genes leads to conversion of heme to an intermediate biliverdin for phycocyanobilin synthesis, and reduces the accumulation of biliverdin in the process of the phycocyanobilin synthesis. The genome of E. coli is further engineered to overexpress related genes of a metabolic pathway of phycocyanobilin, and a strain of recombinant E. coli with high yield of phycocyanobilin is obtained. The recombinant E. coli strain is cultured for 36 hr in a system using glycerol as a substrate, and the phycocyanobilin yield can reach 147 mg/L.

Abstract: Disclosed is recombinant Escherichia coli for 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 E. coli, then heterologously expresses fpk from Bifidobacterium adolescentis, expresses the endogenous genes ppsA and tktA of E. coli, and then expresses aroGfbr and tyrAfbr. 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.

Abstract: The present disclosure provides a P450 cytochrome enzyme for andrographolide synthesis and its application, belonging to the field of bioengineering. The present disclosure uses Saccharomyces cerevisiae CEN.PK2-1D as a host, and implements knockout of ROX1 and GAL80 genes on the genome, and integrative expression of GGPP synthase encoding gene and CPS diterpene synthase encoding gene at ROX1 site; and implements free expression of ApCPR and CYP71A8 and CYP71D10 both with truncated signal peptides, successfully constructing recombinant S. cerevisiae, and achieving de novo synthesis of 3,15,19-Trihydroxy-8(17),13-ent-labdadiene-16-oic acid. Compared with the blank, a response value of a product peak reaches 1.9*106, and this strategy provides necessary reference for analyzing biosynthetic pathway of andrographolide and using metabolic engineering to synthesize andrographolide and related derivatives thereof.

Abstract: The present disclosure discloses a recombinant Escherichia coli for producing rosmarinic acid and application thereof, belonging to the technical fields of genetic engineering and bioengineering. In the present disclosure, FjTA derived from Flavobacterium johnsoniae, endogenous hpaBC derived from E. coli, CbRAS derived from Coleus blumei, HPPR derived from Coleus scutellarioides, and Pc4CL1 derived from Petroselinum crispum are heterologously expressed in E. coli, realizing synthesis of rosmarinic acid. TcTAL derived from Trichosporon cutaneum and tyrC for removing feedback inhibition are introduced, further increasing synthesis throughput of caffeic acid, and PmLAAD derived from Proteus myxofaciens is heterologously expressed, realizing redistribution of L-DOPA. An endogenous gene menl is knocked out, improving the content and stability of a rosmarinic acid precursor. The recombinant strain constructed in the present disclosure can produce rosmarinic acid by fermentation at a yield of up to 511.

Abstract: 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 recombinant S. cerevisiae CA-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 S. cerevisiae. The recombinant S. cerevisiae can 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 ?g/L by optimizing fermentation conditions, and the fermentation time is shortened significantly.

Abstract: 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 recombinant S. cerevisiae CA-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 S. cerevisiae. The recombinant S. cerevisiae can 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 µg/L by optimizing fermentation conditions, and the fermentation time is shortened significantly.

Abstract: 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.

Abstract: The present disclosure relates to a method for improving the yield and production intensity of Gluconobacter oxydans (G. oxydans) 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 G. oxydans, 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 G. oxydan-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·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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: The present disclosure relates to a method for improving the yield and production intensity of Gluconobacter oxydans (G. oxydans) 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 G. oxydans, 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 G. oxydan-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·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.

Abstract: The invention provides a high-throughput screening system based on a single manipulator, and belongs to the field of biotechnology and detection equipment. A high-throughput screening system based on single manipulator, comprises of one manipulator which is surrounded by deep well plate loading platform, shallow well plate loading platform, pipetting head loading platform, control cabinet, waste pipetting heads container platform, shallow well plate recycling platform, deep well plate recycling platform, Microplate Reader with oscillating mixer and items-placing platform in a counterclockwise or clockwise order. The present invention is a combination of microbiology and mechanics. It promotes the automation of microbial high-throughput screening system for screening microorganisms with specific properties. It effectively contributes to the development of microbial breeding technology and further promotes the development of microbial science.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: The present invention provides methods for enhancing ?-KG production in Yarrowia lipolytica, relates to the field of metabolic engineering. This invention successfully overexpresses the glutamate dehydrogenase in wild type strain Y. lipolytica WSH-Z06 to construct the recombinant Y. lipolytica WSH-Z06 which regulates the glutamate catabolism to synthesis ?-KG. L-methionine imine is added into the fermentation medium during the process to strengthen the supply of intracellular glutamate and inhibite the intracellular glutamine synthesis from glutamate metabolism and then enhance the accumunation of ?-KG. Therefor, the present invention provides an effective method for enhancing the accumunation of ?-KG through regulation of intracellular amino acid metabolism.