143 results about "Recombinant yeast" patented technology

The present invention relates to a recombinant yeast with high squalene production, a construction method and a use thereof, wherein the recombinant yeast is a saccharomyces cerevisiae that expresses or overexpresses HMGR gene, ADA gene, LEU2 gene, MVA pathway gene and squalene synthase gene ERG9. The present invention integrates acetaldehyde dehydrogenase gene ADA and HMG-CoA reductase gene tHMGR in multiple copies in saccharomyces cerevisiae, integrates the screened high-activity phosphoketolase gene PK and phosphotransacetylase gene PTA, weakens epoxysqualene synthesis, strengthens the mevalonate pathway, overexpresses yeast endogenous squalene synthase gene, etc. to obtain a recombinant yeast with high squalene production, so that the production in the shake flask stage of the production process exceeds 5g / L, and the production in the fed batch fermentation stage exceeds 50g / L, breaking through the current bottleneck of microbial synthesis of squalene production, and is suitable for practical promotion and application.

The present disclosure provides a recombinant yeast host cell having a plurality of native and / or heterologous enzymes that function in an engineered metabolic pathway to convert fructose-6-phosphate to acetyl-coenzyme A. The recombinant yeast host cell can be used, for example, in a process for making acetyl-coenzyme A, acetone and / or isopropanol. The plurality of native and / or heterologous enzymes is activated, upregulated, or overexpressed. The plurality of native and / or heterologous enzymes comprises a phosphoketolase; and optionally an acetate kinase; and / or a phosphotransacetylase. The recombinant yeast host cell comprises at least one of: at least two copies of a heterologous nucleic acid molecule encoding the phosphoketolase; a native and / or heterologous enzyme that functions in an engineered non-oxidative pentose phosphate pathway, wherein the native and / or heterologous enzyme is activated, upregulated, or overexpressed; and / or a native and / or heterologous protein that functions in an engineered metabolic pathway to convert pantothenate into acetyl-coenzyme A, wherein the native and / or heterologous protein is activated, upregulated or overexpressed and comprises at least one of: FEN2 or CAB1.

The invention provides recombinant yeast for producing bakuchiol as well as a construction method and application of the recombinant yeast. Wherein the recombinant yeast is Yarrowia lipolytica containing isopentenyl transferase, and the recombinant yeast is a Yarrowia lipolytica containing isopentenyl transferase. Wherein a gene for coding the isopentenyl transferase is derived from Psoralea coryllifelia Linn (Psoralea coryllifelia Linn), and the isopentenyl transferase is used for coding the isopentenyl transferase. By utilizing the recombinant yeast provided by the invention, the bakuchiol with high yield can be efficiently and quickly prepared through a biosynthesis method, and a technical support is provided for subsequent industrialization of the bakuchiol.

The present disclosure provides a recombinant yeast host cell for converting a carbohydrate to ethanol and acetone. The recombinant yeast host cell comprises an engineered metabolic pathway to convert acetyl-CoA to acetone, a downregulated native alcohol dehydrogenase (ADH) gene, and a heterologous alcohol dehydrogenase.

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.

The invention relates to the technical field of biological fermentation, in particular to a recombinant yeast cell and application thereof in production of thaumatin. The recombinant yeast cell contains nucleic acids corresponding to thaumatin, PDI and MSN4 genes, and can functionally express the thaumatin, PDI and MSN4 genes. The invention provides the recombinant yeast cell for co-expressing the chaperonin PDI and the transcription factor MSN4 to improve the secretory expression of the thaumatin, and accidentally, under the synergistic effect of the chaperonin PDI and the transcription factor MSN4, the secretory expression quantity of the exogenous thaumatin is remarkably increased by 3.25 times.

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.

The present invention belongs to the field of bioengineering, genetic engineering technology and oligosaccharide (rare sugar) synthesis technology, and specifically relates to a method for producing lactose-N-disaccharide using genetically modified recombinant yeast. The technical scheme of the present invention is: a recombinant yeast cell is introduced with (1) a heterologous lactose-N-disaccharide phosphorylase ( LNBP ) gene, and (2) fructose-1-phosphate phosphorylase ( YQAB ) gene, and further preferably, also includes (3) destruction or modification of a marker gene. The recombinant yeast cell has the ability to synthesize lactose-N-biose (Lacto-N-biose, LNB) from lactose, and the yield of LNB is greatly improved by a molecular switch.

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 present invention belongs to the field of fermentation engineering, and specifically relates to a yeast and its application in linseed oil fermentation and a linseed hydroponic oil product. The genetically recombinant yeast expresses phenylalanine deaminase, tyrosine deaminase, cinnamic acid hydroxylase, 4-coumaric acid CoA ligase, chalcone synthase, chalcone isomerase, flavonoid 3-hydroxylase and stilbene synthase. The linseed hydroponic oil obtained by fermenting genetically recombinant yeast and linseed oil in the present invention has enhanced stability and is not prone to mutation; and the linseed hydroponic oil has a higher total flavonoid content, better antioxidant activity and tyrosinase inhibition activity than linseed oil.

The invention relates to the technical field of gene engineering, in particular to a method for reducing acetic acid accumulation in the process of ethanol-induced expression of alpha-lactalbumin by regulating pichia pastoris. According to the recombinant yeast strain provided by the invention, the accumulation amount of acetic acid is greatly reduced in the process of inducing expression of target protein by adopting ethanol, acetic acid can be basically not accumulated, relatively high biomass and target protein yield can be achieved, and the highest yield of human alpha-lactalbumin up to now is achieved; and a strain resource and a foundation are provided for application of an ethanol-induced pichia pastoris expression system and development of bioengineering milk protein. The invention provides a method for solving the problem of acetic acid accumulation of pichia pastoris in the process of inducing expression of the target protein by using ethanol, and the method provides an effective strategy for popularization and application of an ethanol-induced expression system and improvement of the yield of the target protein induced and expressed by using the ethanol. The method has important significance on production of proteins such as alpha-lactalbumin and the like.

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 present invention relates to a recombinant yeast characterized in that its genome comprises a metabolite-inducible promoter in place of the native promoter of the AGA1 gene, to a plasmid comprising (i) a metabolite-inducible promoter operatively linked the nucleotide sequence coding for a fusion protein comprising a protein of interest, and to the combined use of said yeast and plasmid in a Yeast Surface Display method.

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 invention belongs to the technical field of molecular biology, and particularly relates to a recombinant vector for expressing OrfV truncated F1L protein, a strain and application of the recombinant vector. The preparation method comprises the following steps: truncating an F1L gene of an orf virus, introducing a His tag and a termination codon sequence, and constructing into a pYD1 plasmid to obtain a pYD1-F1L recombinant plasmid; and introducing the recombinant plasmid into competent cells of saccharomyces cerevisiae to obtain the recombinant yeast strain. According to the invention, the F1L gene of OrfV is truncated, so that the immunogenicity of the OrfV is effectively retained; the anchoring protein is displayed on the surface of the saccharomyces cerevisiae, so that the immune function of the saccharomyces cerevisiae can be developed; recombinant saccharomyces cerevisiae is adopted as a delivery carrier, the antigen protein can better play an immune role by means of the constant value of the recombinant saccharomyces cerevisiae in intestinal tracts, and meanwhile, the risk that the F1L protein is degraded by protease when being orally taken is further reduced. The invention provides a new direction for the development of OrfV oral vaccines.

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.