Abstract: The present invention pertains to a method for preparing ultra-thin RhCu nanostructures modified with M-TPP and applications thereof. By adjusting the feeding ratio of raw materials, the morphology, composition, purity, and size of RhCu M-tpp can be customized. The simplicity of operation and the robustness of reaction parameters contribute to high reproducibility of the target products. Furthermore, large-scale production is achievable by proportionally increasing the concentrations of metal ion precursors, reducing agents, and surfactants, demonstrating significant potential for industrial-scale production of molecule-modified ultra-thin RhCu nanostructures.

Abstract: The disclosure discloses a Saccharomyces cerevisiae lysate containing one or more cytokines, a preparation method therefor and an application thereof in cell-cultured meat, and belongs to the technical field of genetic engineering and cell-cultured meat. The disclosure provides a method for further increasing recombinant cytokine yields using the GRAS strain of S. cerevisiae recombinantly expressing a single cytokine or co-expressing a combination of cytokines through promoter optimization, knockout of endogenous protease in yeast, genome-integrated expression and other means. The yeast lysate may be directly used for muscle stem cell culture after the process such as filtration, sterilization, and cytokine concentration measurement, effectively promoting muscle stem cell proliferation. The disclosure avoids the complex purification process in the production of recombinant cytokines, reduces the cost of cytokine production, and provides new ideas for large-scale low-cost development of cell-cultured meat.

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: 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: 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: The present disclosure provides myoglobin (MB) and an expression vector and an expression engineering bacterium thereof, and use thereof, and relates to the technical field of genetic engineering. In the present disclosure, recombinant Escherichia coli strains with signal peptides Pel B and Omp A inserted under same conditions have an expression level of Sus scrofa myoglobin (SsMB) increased by 2.06 and 1.17 times, respectively, compared with an original expression strain of the SsMB. The signal peptides that can increase the expression level of the SsMB provide a new idea for research and application of improving the expression level of the SsMB.

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 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 P. pastoris. On this basis, optimization of the medium and culture conditions of recombinant P. pastoris can 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%.

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 invention relates to the technical field of bioengineering and provides a method for improving the specificity and affinity of an aptamer. The method includes: S1, screening a target of an aptamer from a compound information database by virtual computing; S2, verifying the screening result in Step S1 through experiments; S3, performing virtual saturation mutation on a site of the aptamer, and screening out a mutation site of the aptamer; S4, performing base substitution to the mutation site of the aptamer; and S5, detecting the binding parameter of the aptamer after base substitution with the target screened in Step S1, and selecting an aptamer with improved specificity and affinity after base substitution. An efficient molecular design-guided method is developed by computer rational calculation, to improve the specificity and binding affinity of the aptamer by directional modification. The present invention is of great significance for the practical application of aptamers.

Abstract: 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 P. pastoris. On this basis, optimization of the medium and culture conditions of recombinant P. pastoris can 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%.

Abstract: The present disclosure discloses a visualized screening method for an Aspergillus recombinant strain with multigene editing and belongs to the technical field of gene engineering. CRISPR-Cas9 is used in the disclosure to cleave spore color change-related genes and a target gene in Aspergillus at the same time, such that editing of the target gene is visualized and an Aspergillus niger strain with multigene editing can be rapidly and efficiently screened out through spore phenotypes. Through different combinations of visualized genes and non-phenotypic change genes, rapid screening of the strain with multigene editing and simultaneous screening of multiple visualized genes are realized, and use of resistance genes in industrial strains is reduced.

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 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.