108 results about "Recombinant yeast" patented technology

The present invention provides a yeast recombinant type XVII humanized collagen and its preparation method, in particular, a recombinant humanized collagen expressing the 15th helical region of type XVII collagen, a recombinant engineering bacterium and its preparation method, belonging to the technical fields of genetic engineering and synthetic biology. The present invention provides the full-length sequence of the 15th helical region of type XVII collagen, and mutants are obtained by mutating amino acid residues based on this sequence; for the first time, the present invention expresses the full-length sequence of the 15th helical region of type XVII collagen in Pichia pastoris, establishes a complete set of fermentation processes and purification processes, improves the protein expression level and reduces degradation, and through experiments, it is verified that the obtained protein has biological activities such as cell adhesion activity and cell migration promotion activity that reach or exceed those of natural human collagen, can achieve the purpose of real product application, and can be widely applied in the fields of medicine, medical devices, biological materials, tissue engineering, cosmetics, etc.

A recombinant yeast cell that functionally expresses: —a nucleic acid sequence encoding a protein having glycerol dehydrogenase activity; —a nucleic acid sequence encoding a protein having dihydroxyacetone kinase activity; and —a nucleic acid sequence encoding a protein having glycerol transporter activity, wherein the expression of the nucleic acid sequence encoding the protein having glycerol transporter activity is under control of a promoter (the “GT promoter”), which GT promoter has an anaerobic / aerobic expression ratio for the glycerol transporter of 2 or more, and a process for the production of ethanol using such recombinant yeast cell.

The invention discloses saccharomyces cerevisiae recombinant bacteria for efficiently producing terpenoids as well as a construction method and application of the saccharomyces cerevisiae recombinant bacteria. According to the construction method, saccharomyces cerevisiae rich in terpenoid precursor farnesyl pyrophosphate is taken as a starting strain, the GAL80 protein level in a galactose induction system is regulated and controlled by introducing an N-Degron strategy, and meanwhile, a glucose concentration response type promoter HXT1 is introduced to effectively regulate and control the expression of GAL80 from the transcriptional level; the saccharomyces cerevisiae recombinant strain for efficiently producing the terpenoids can be obtained. The strategy is applied to a saccharomyces cerevisiae engineering strain of beta-elemene and patchouli alcohol, and a remarkable effect is achieved. And in the high-density fermentation process of the saccharomyces cerevisiae, the yield of the beta-elemene is obviously increased and reaches 4818.6 mg / L. The invention provides a solid research basis for sustainable biosynthesis of terpenoids, and has a wide application prospect.

A recombinant yeast cell functionally expressing: a) a nucleic acid sequence encoding a protein comprising phospho-ketolase (PKL) activity (EC 4.1.2.9 or EC 4.1.2.22) and / or a nucleic acid sequence encoding a protein having phosphotransacetylase (PTA) activity (EC 2.3.1.8) and / or a nucleic acid sequence encoding a protein having acetate kinase (ACK) activity (EC 2.7.2.12); and / or) a nucleic acid sequence encoding a protein having transketolase activity (EC 2.2.1.1), wherein the expression of the nucleic acid sequence encoding the protein having transketolase activity is under control of a promoter (the “TKL promoter”), which TKL promoter has an anaerobic / aerobic expression ratio for the transketolase of (2) or more.

The invention provides a preparation method of 11-seed fermentation oil and fermentation liquor with multiple effects, and belongs to the technical field of fermentation. The method comprises the following steps: S1, preparing recombinant saccharomycetes; s2, jointly fermenting lactobacillus, recombinant saccharomycetes and 11 seed oil; s3, separating an oil phase and a water phase, and taking the oil phase; and S4, dehydrating the oil phase to obtain the 11-seed fermentation oil. And S5, further refining the water phase to obtain the 11-seed fermentation liquor. The 11-seed fermented oil prepared by transforming a microbial metabolic network is rich in dihydroquercetin, and has multiple effects of resisting oxidation, resisting wrinkles, tightening, repairing cells and the like. In the fermentation process of the 11-seed fermentation oil, 11-seed fermentation liquor can be obtained, and the fermentation liquor has multiple effects of resisting oxidation, relieving, moisturizing, resisting wrinkles, tightening and the like.

The present invention relates to treatment of COVID-19 and methods thereof. A vaccine composition for inducing immunity against coronavirus in a subject comprises a recombinant nucleic acid encoding an N-ETSD, which is a modified nucleocapsid protein comprising an endosomal targeting sequence, and / or a recombinant nucleic acid encoding an S-fusion, which is a modified spike protein with improved surface expression. The vaccines can be formulated as recombinant nucleic acids, recombinant yeasts, and / or recombinant viruses, such as adenoviruses, and can be administered via injection and / or mucosal delivery.

The invention discloses a recombinant yeast for expressing large yellow croaker iridovirus MCP protein and a preparation method and application of the recombinant yeast in preparation of vaccines, and is characterized in that the recombinant yeast is saccharomyces cerevisiae transformed with pYD1-MCP recombinant plasmids and is named as EBY100 / pYD1-MCP, the pYD1-MCP recombinant plasmids can express the MCP protein on a yeast surface display system, the nucleotide sequence of a gene segment of the MCP protein is shown as SEQ ID NO.1, the nucleotide sequence of the gene segment of the MCP protein is shown as SEQ ID NO.2, and the nucleotide sequence of the gene segment of the MCP protein is shown as SEQ ID NO.2. The preparation method comprises a step of constructing a recombinant plasmid pYD1-MCP and a step of constructing a recombinant yeast EBY100 / pYD1-MCP, the oral vaccine of the larimichthys crocea iridovirus vaccine can be prepared, and the oral vaccine has the advantages of being high in protection rate, safe, low in cost, convenient to use and good in application prospect in prevention and control of the larimichthys crocea iridovirus.

A recombinant yeast cell functionally expressing: a) a nucleic acid sequence encoding an enzyme having NADH-dependent nitrate reductase activity and / or a nucleic acid sequence encoding an enzyme having NADH-dependent nitrite reductase activity; and b) a nucleic acid sequence encoding a protein having transketolase activity (EC 2.2.1.1), wherein the expression of the nucleic acid sequence encoding the protein having transketolase activity is under control of a promoter (the “TKL promoter”), which TKL promoter has an anaerobic / aerobic expression ratio for the transketolase of 2 or more.

The invention discloses an optimized recombinant chicken double-cell factor as well as a preparation method and application thereof. The chicken recombinant double-cell factor coding gene provided by the invention contains complete ChIL-2 and ChIFN-alpha structures. The preparation process comprises the following steps: selecting a pichia pastoris expression system; optimizing the recombinant chicken double-cell factor according to the codon preference of pichia pastoris; inserting the optimized gene sequence into an expression vector, and introducing host bacteria to construct a recombinant yeast engineering strain; carrying out inoculated culture by using the obtained recombinant yeast, and screening strains containing target protein based on expression level to obtain required culture bacteria; and breeding and culturing the obtained high-expression-level culture bacteria. According to the invention, ChIL-2 and ChIFN-alpha are connected through a modified flexible linker, and a nucleotide sequence is replaced by preferred codons of pichia pastoris, so that a chicken recombinant cell factor can be secreted to a supernatant, the defect of ChIL-2 expression quantity is overcome, and the problem that proteins of ChIL-2 and ChIFN-alpha mainly exist in bacteria and inclusion bodies in prokaryotic expression is solved.

The invention discloses zingiberone epoxidase protein in ligustrum sinense as well as a biological material and application of the zingiberone epoxidase protein. The amino acid sequence of the zingiberone epoxidase protein CYP76S110 to be protected in the ligustrum sinense is 2 in a sequence table, and the coding nucleotide sequence of the zingiberone epoxidase protein CYP76S110 is 1 in the sequence table. According to the embodiment of the invention, after the coding gene of the CYP76S110 protein is introduced into yeast for expression, an obtained recombinant yeast microsome solution containing the CYP76S110 protein can be used for catalyzing zingiberone to synthesize zingiberone epoxide (zingiberone epoxide), and the recombinant yeast microsome solution can be used for catalyzing zingiberone to synthesize zingiberone epoxide (zingiberone epoxide). The method can be applied to in-vitro biosynthesis of the zingiberone derivative.

An object of the present invention is to provide a recombinant yeast that can highly express a target protein in a tightly regulated manner without using methanol, and to provide a method for producing a target protein using the recombinant yeast, a kit for producing the recombinant yeast, and a method for producing the recombinant yeast. According to the present invention, there is provided a recombinant yeast containing a first exogenous sucrose-inducible promoter for expressing a target protein.

The present invention relates to a yeast cell, in particular a recombinant yeast cell, the cell lacking enzymatic activity needed for the NADH-dependent glycerol synthesis or the cell having a reduced enzymatic activity with respect to the NADH-dependent glycerol synthesis compared to its corresponding wild-type yeast cell, the cell comprising one or more heterologous nucleic acid sequences encoding an NAD+-dependent acetylating acetaldehyde dehydrogenase (EC 1.2.1.10) activity. The invention further relates to the use of a cell according to the invention in the preparation of ethanol.

The present invention discloses a recombinant saccharomyces cerevisiae for efficiently biosynthesizing inositol and its construction method and application, and belongs to the field of microbial genetic engineering. The recombinant saccharomyces cerevisiae of the present invention strengthens the expression of hexokinase gene hxk, inositol 3-phosphate synthase gene ino, inositol monophosphatase gene suhb, and realizes the rearrangement of glucose metabolism by knocking out the transcriptional repressor opi1 of inositol 3-phosphate synthase (Ino1) and glucose 6-phosphate dehydrogenase gene zwf1, and relieves feedback inhibition; In addition, the present invention further increases the yield of inositol by screening inositol external emission protein gene ompf, ompc and knocking out inositol internal transport protein gene itr1, itr2. The present invention realizes the efficient production of inositol (yield is as high as 195.7g / L), with good industrialization prospect.

The invention provides a human FGFR3 (Fibroblast Growth Factor Receptor 3) antigen expression recombinant yeast vaccine as well as a construction method and application thereof. A specific primer is designed to amplify an FGFR3 gene, a pEGKT316-FGFR3 recombinant plasmid is constructed, a saccharomyces cerevisiae YEF473A strain is transformed, and the vaccine is prepared through induced expression and thermal inactivation. The vaccine can effectively activate DC cell maturation (CD80 + cell proportion reaches 78.7%) and promote Th1 type cell factor secretion (IL-12p70 reaches 5889pg / mL), and shows a remarkable tumor inhibition effect (plt; 0.05), and a novel specific treatment means is provided for postoperative immunotherapy of bladder cancer.

The present disclosure describes a recombinant yeast host cell having a first engineered metabolic pathway to convert fructose-6-phosphate (F-6-P) into acetyl-coenzyme A. The first engineered metabolic pathway comprises: a genetic modification for expressing a heterologous phosphoketolase capable of converting fructose-6-phosphate into erythrose-4-phosphate, and at least one genetic modification for decreasing native 6- phosphofructo-2-kinase activity classified under Enzyme Commission No. 2.7.1.105. Optionally, the recombinant yeast host cell comprises a second engineered metabolic pathway for convert acetyl-coA into a fermentation product. The present disclosure further describes a process for making a fermentation product. The process comprises contacting the recombinant yeast host cell described herein with a carbohydrate source under a condition allowing the conversion of at least a part of the carbohydrate into a fermentation product.

The present disclosure concerns a process for fermenting a biomass with a reduced dose of a purified exogenous enzyme (which can be, for example a purified exogenous glucoamylase). The process comprises contacting a biomass (which may comprise starch) with a recombinant yeast host cell. The recombinant yeast host cell has a genetic modification for expressing a heterologous polypeptide having starch or dextrin hydrolase activity (which may be, for example, from a glucoamylase). The nucleic acid molecule encoding the heterologous polypeptide having starch or dextrin hydrolase activity comprises allowing the secretion of the heterologous polypeptide.

The present disclosure concerns recombinant yeast host cells expressing cell-associated heterologous food and / or feed enzymes which are expressed during the propagation phase of the recombinant yeast hosts cells. The recombinant yeast host cells can be used in a subsequent production process to make food and / or feed products, for example, baked products.

The present invention relates to a yeast cell, in particular a recombinant yeast cell, the cell lacking enzymatic activity needed for the NADH-dependent glycerol synthesis or the cell having a reduced enzymatic activity with respect to the NADH-dependent glycerol synthesis compared to its corresponding wild-type yeast cell, the cell comprising one or more heterologous nucleic acid sequences encoding an NAD+-dependent acetylating acetaldehyde dehydrogenase (EC 1.2.1.10) activity. The invention further relates to the use of a cell according to the invention in the preparation of ethanol.

The invention belongs to the technical field of gene recombination, and particularly relates to a recombinant human III-type triple helix collagen, a recombinant yeast strain for expressing the recombinant human III-type triple helix collagen and a construction method of the recombinant human III-type triple helix collagen. The amino acid sequence of the recombinant human III-type triple-helix collagen is shown as SEQ ID NO. 1, and the recombinant human III-type triple-helix collagen has triple-helix conformation close to a natural biological structure of collagen and has good application value and potential. According to the recombinant yeast strain disclosed by the invention, human-derived III-type collagen and proline hydroxylase are co-expressed in yeast cells without YPS1 protease, and meanwhile, a gene beneficial to vesicle transport and protein folding is transferred, so that hydroxylation of the recombinant collagen can be realized, a natural triple helix conformation is formed, and the recombinant yeast strain can be used for preparing a recombinant collagen product. The degradation problem of the collagen in the expression process can be solved, and the extracellular secretion expression of the triple helix collagen is realized.

The present invention discloses a method for producing eucommia gum by fermenting eucommia leaves with recombinant yeast. The main steps of the method include: constructing an expression vector containing the cellulase genes Cel5A / Cel6A of Trichoderma reesei, introducing Saccharomyces cerevisiae to construct an engineered yeast, and using the engineered yeast to further enzymatically hydrolyze the eucommia leaves after ultrasonic pretreatment to achieve complete release of eucommia gum, and using Soxhlet extraction to purify the eucommia gum from the fermentation product to achieve full extraction of the eucommia gum. The method of the present invention does not require high-concentration alkaline and acid solution cracking and repeated rinsing, thus avoiding the destruction of the rubber morphology during the extraction process and reducing the generation of harmful waste liquid during the extraction process. It has guiding significance for achieving green extraction of eucommia gum.

The application discloses a recombinant bacterium and a use thereof, the recombinant bacterium is a yeast containing or expressing aai-na-xiang monoterpene synthase BbTPS3 or an ai-na-xiang monoterpene synthase BbTPS3 fusion protein in vivo, and is used for producing levorotatory borneol; experiments prove that the recombinant bacterium after modification (such as codon optimization, protein truncation, Kozak increase or protein fusion) can improve the yield of levorotatory borneol, and is suitable for industrialized production of levorotatory borneol.

The invention provides a recombinant yarrowia lipolytica strain with high yield of eicosapentaenoic acid as well as a construction method and application of the recombinant yarrowia lipolytica strain, and belongs to the technical field of genetic engineering. The invention provides a recombinant yarrowia lipolytica strain Po1f: Y1 capable of reconstructing an EPA (eicosapentaenoic acid) synthetic route, and the recombinant yarrowia lipolytica strain Po1f: Y2 is constructed by taking the recombinant yarrowia lipolytica strain Po1f: Y1 as a host and overexpressing an LPCAT (Low Performance Protein Amino Transferase) mutant; and knocking out encoding genes such as PEX10, GUT2, TGL4, SNF1 and the like, and combining with overexpressed DGAT1 and the like to obtain a recombinant yarrowia lipolytica strain Po1f: Y3 with high yield of EPA. The recombinant yarrowia lipolytica strain Po1f: Y3 strain EPA accounts for 50% of the relative content of total fatty acid, the oil content accounts for 48% of the weight of stem cells, and the biomass of recombinant yeast in a 7.5 L fermentation tank exceeds 100 g stem cell weight / L.

This invention relates to a genetically engineered bacterium producing OPO-type structural esters, which is a recombinant yeast cell containing the ChDes9-2 gene encoding stearoyl-CoAΔ9 desaturase. Specifically, this genetically engineered bacterium is constructed by replacing the OLE1 gene encoding Δ9 desaturase with the ChDes9-2 gene encoding stearoyl-CoAΔ9 desaturase in Saccharomyces cerevisiae strain YS58, and by overexpressing the GAT1 and / or SLC and / or PAP and / or DGAT genes. This effectively increases the proportion of OPO-type structural esters in the lipids of Saccharomyces cerevisiae, making the content of OPO-type structural esters in triglycerides in the genetically engineered Saccharomyces cerevisiae reach 17.2%, thus overcoming the limitations of OPO-type structural ester sources.

The present invention relates to the field of biotechnology, and specifically to a method for constructing a recombinant yeast with high-yield agroclavine and its application. The present invention optimizes the yield of agroclavine by reconstructing the heterologous synthesis pathway by determining the subcellular expression sites of the enzymes (DmaW, EasF, EasE, EasC, EasD, EasA, EasG) of the heterologous pathway for the biosynthesis of agroclavine in Saccharomyces cerevisiae strains. The present invention determines the subcellular expression sites and expression levels of the enzymes (DmaW, EasF, EasE, EasC, EasD, EasA, EasG) of the heterologous pathway for the biosynthesis of agroclavine in Saccharomyces cerevisiae strains, and further proves that the introduction of this heterologous pathway will cause cofactor imbalance in the endogenous environment, providing favorable evidence for subsequent further optimization of agroclavine production, and also providing a high-yield chassis for the biosynthesis of ergotic acid derivatives using agroclavine as a precursor.

The present invention relates to the field of genetic engineering technology, and in particular to a method for regulating Pichia pastoris to reduce the accumulation of acetic acid during the ethanol-induced expression of α-lactalbumin. The recombinant yeast strain provided by the present invention has a greatly reduced accumulation of acetic acid during the expression of a target protein by ethanol induction, and can achieve essentially no accumulation of acetic acid, thereby achieving a higher biomass and target protein yield, and achieving the highest yield of human α-lactalbumin to date. This provides strain resources and a foundation for the application of the ethanol-induced Pichia pastoris expression system and the development of bioengineered milk proteins. The present invention provides a method for solving the problem of acetic acid accumulation in the process of Pichia pastoris expressing a target protein by ethanol induction. This method provides an effective strategy for the promotion and application of the ethanol-induced expression system and for improving the yield of the target protein expressed by ethanol induction, and is of great significance for the production of proteins such as α-lactalbumin.

Recombinant yeasts bioengineered to overexpress genes for utilization of cellulosic and hemicellulosic fibers and / or produce lipid, and methods of use thereof. The yeasts are modified to express, constitutively express, or overexpress an exo-cellobiohydrolase I, an exo-cellobiohydrolase II, an endoglucanase, an endoxylanase, or any combination thereof. The yeasts may also be modified to express, constitutively express, or overexpress a lytic polysaccharide monooxygenase, an arabionofuranosidase, a xylanases, an acetylxylan esterase, or any combination thereof. The methods include cultivating the recombinant yeasts to convert low value organic stillage byproducts such as cellulose and hemicellulose into lipids suitable for biodiesel production and other higher value uses.

The present invention relates to a recombinant yeast for high-level production of adipic acid, wherein the activity of endogenous acyl-CoA oxidase is weakened compared to the inherent activity, and the activity of plant-derived acyl-CoA oxidase is enhanced compared to the inherent activity. The recombinant yeast according to the present invention can produce adipic acid at a high yield compared to existing unmodified microorganisms.

A recombinant yeast for producing lacto-N-triose, and an application thereof, belonging to the bioengineering field wherein the recombinant yeast includes an amino acid sequence of a β-N-acetylglucosaminyl transferase and a coding nucleic acid sequence thereof, and / or a disruption of a marker gene. The technical solution of the recombinant yeast has the advantages of easy availability of raw materials, simple and feasible method, suitable for industrial production, and excellent application prospects.

The invention relates to the technical field of microorganisms, and discloses recombinant yeast for efficiently synthesizing a GPP derivative compound as well as a construction method and application of the recombinant yeast. The recombinant yeast strain takes a yeast strain as an original strain, and is obtained by introducing a target coding gene into a cell nucleus; the target coding gene comprises a coding gene of a mevalonic acid (MVA) synthetic pathway enzyme, a coding gene of a farnesyl diphosphate synthase mutant and a coding gene of optional geranyl diphosphate derivative synthase. According to the invention, an MVA pathway and a key enzyme are introduced into a cell nucleus, so that competitive consumption of a sterol synthesis pathway in cytoplasm on geranyl diphosphate (GPP) is effectively avoided, and efficient synthesis of the GPP derivative compound is realized. Besides, the recombinant yeast strain disclosed by the invention can be combined with a reported peroxisome platform to construct a double-organelle platform strain, so that the yield of GPP derivative compounds can be further increased.

The present invention relates to a recombinant microorganism, preferably a recombinant yeast, capable of producing caffeic acid comprising a heterologous gene coding for an enzyme of the hydrolase family capable of breaking, preferably of hydrolyzing, the caffeoyl-shikimate bond to produce caffeic acid from caffeoyl-shikimate. Said microorganism, preferably said recombinant yeast, may also be capable of producing ferulic acid from the caffeic acid obtained. The present invention also relates to a method for producing caffeic acid and a method for producing caffeic acid and / or ferulic acid, using microorganisms, preferably yeasts, according to the invention. Finally, the invention also relates to the use of microorganisms, preferably yeasts, according to the invention to produce caffeic acid and / or ferulic acid.