66 results about "Recombinant yeast" patented technology

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 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.

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.

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 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 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.

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.

Described herein are recombinant Kluyveromyces lactis (K. lactis) yeasts which are capable of the highly efficient expression of one or more foreign proteins and are suitable for use as a vaccine for generating a protective immune response against pathogens. The invention provides in particular K. lactis strains for the targeted cloning of foreign antigen-coding nucleic acids into the yeast genome of the K. lactis strain, which is characterized in that the K. lactis strain has integrated expression cassettes for foreign antigens as an alternative or in addition to the KILAC4 locus on the KIURA3-20 locus (KLLA0E22771g) and / or on the KIMET5-1 locus (KLLA0B03938g). The invention further relates to integrative expression vectors and to methods for producing the K. lactis strains of the invention as well as to the use thereof as vaccines.

This invention relates to the field of biological products, and particularly to the preparation and application of hangover relief products. This invention provides an alcohol dehydrogenase mutant, an aldehyde dehydrogenase mutant, an expression vector, a combination of recombinant yeast strains, and yeast single-cell oral capsules. By cloning the alcohol dehydrogenase and aldehyde dehydrogenase genes into the *Saccharomyces cerevisiae* expression plasmid pYX212, and then transforming the *Saccharomyces cerevisiae* into the *Saccharomyces cerevisiae*, engineered yeast is obtained. Utilizing the protective effect of yeast cells, oral administration is achieved, effectively reducing the destruction of recombinant enzymes by gastric juice. The yeast single-cell oral capsules can accelerate alcohol metabolism after daily drinking and promote hepatocyte regeneration; they are convenient and quick to take and have significant effects.

The invention discloses a recombinant yeast engineering bacterium for secretory expression of recombinant bovine lactoferrin as well as a construction method and application of the recombinant yeast engineering bacterium, and belongs to the technical field of genetic engineering. According to the invention, a CRISPR / CAS9 gene editing technology is utilized to construct sgRNA expression vectors and bovine lactoferrin homologous recombination gene expression cassettes targeting multiple sites of yeast genomes AOX1, AOX2, FLD1, DAS1 and DAS2, and the sgRNA expression vectors and the bovine lactoferrin homologous recombination gene expression cassettes are transformed into yeast host bacteria to obtain recombinant yeast engineering bacteria with bovine lactoferrin genes inserted at multiple sites. The yield of the bovine lactoferrin of the recombinant yeast engineering bacterium is remarkably increased, heterologous secretion and efficient expression of the bovine lactoferrin in pichia pastoris are achieved, industrial production and construction of the bovine lactoferrin are promoted, and the recombinant yeast engineering bacterium has wide application value.

The present disclosure concerns the use of specific genetic modification(s) for improving sulfite tolerance in recombinant yeast host cells. The genetic modification(s) is (are) designed to allow the expression of an heterologous transcription factor favoring the expression of a SSU1 polypeptide and / or the expression of an heterologous SSU1 polypeptide in the recombinant yeast host cell(s).

The invention discloses a chaetomium globosum sourced ergothioneine synthetase CgEgt2 and a coding gene cgegt2 of the chaetomium globosum sourced ergothioneine synthetase CgEgt2. The amino acid sequence and the nucleotide sequence of the chaetomium globosum sourced ergothioneine synthetase CgEgt2 are shown as SEQ ID NO.2 and SEQ ID NO.1. The invention further discloses a preparation method of the The invention provides a Kluyveromyces marxianus recombinant strain KMNC-WZ02 containing a cgegt2 gene and an ncegt1 gene, the preservation number of the Kluyveromyces marxianus recombinant strain KMNC-WZ02 is CGMCC (China General Microbiological Culture Collection Center) NO.29599, a two-copy strain of the KMNC-WZ02 is constructed, and the ergothioneine yields of the KMNC-WZ02 and the two-copy strain of the KMNC-WZ02 respectively reach 466 mg / L and 590.3 mg / L after the KMNC-WZ02 and the two-copy strain of the KMNC-WZ02 are fermented for 7 days. The method provided by the invention is of great significance to excavation of EGT synthetases from different sources and construction of efficient EGT production strains.

The invention discloses a Phmul.002G015100 gene for improving saline-alkaline resistance of kidney beans and application of the Phmul.002G015100 gene, and relates to the technical field of biology. The saline-alkaline tolerance key gene Phmul.002G015100 is successfully excavated from kidney beans through a transcriptome sequencing technology, and transgenic yeast verification experiments prove that the gene can remarkably improve the saline-alkaline tolerance of receptor organisms. In a mixed saline-alkali stress culture medium, the growth state of recombinant yeast into which the Phmul.002G015100 gene is introduced is obviously superior to that of control yeast of an empty vector, which indicates that the gene can effectively enhance the tolerance of organisms to saline-alkali stress. The discovery of the gene not only enriches a plant saline-alkaline tolerance gene resource library, provides a key target for analyzing a saline-alkaline tolerance molecular mechanism of the kidney beans, but also provides a core functional gene for saline-alkaline tolerance genetic improvement of the kidney beans.

A construction method of double immobilized recombinant yeast engineering bacteria, engineering bacteria, a catalyst and an application in synthesis of rebaudioside. In the present invention, yeast is used as the chassis cell, an ordered self-assembled multi-enzyme cascade surface display system is constructed, and carrier-free immobilization is coupled with uridine piphosphate glucose (UDPG) in-situ regeneration technology, resulting in a novel yeast whole cell catalyst with double immobilization of enzymes and bacteria. The catalyst uses cheap steviol glycoside (St) and rebaudioside A (Reb A) as substrates, and can synthesize rebaudioside (Reb D / M) in an efficient and economical one-pot method without additional addition of expensive UDPG. The synthesis process does not require cumbersome operations including cell disruption, enzyme separation, purification and immobilization, and avoids bottleneck problems in the related art including material transmembrane transport resistance, mass transfer resistance and product hydrolysis by intracellular enzyme. The catalyst has reusability, and can realize continuous and high-intensity rebaudioside biosynthesis through strain rejuvenation, which provides strong technical support for its industrial application.

The invention belongs to the technical field of recombinant proteins, and particularly relates to a gene for coding a split-ring loganin synthase, the split-ring loganin synthase, a recombinant plasmid, a recombinant bacterium and an application of the split-ring loganin synthase in catalyzing loganin acid to generate a split-ring iridoid compound, wherein the gene is cloned and separated from a gene in gentiana macrophylla pall. The sequence of the gene for coding the split-ring loganin synthase is as shown in SEQ ID NO.01, and the amino acid sequence of the split-ring loganin synthase is as shown in SEQ ID NO.02. The recombinant plasmid contains a sequence shown in SEQ ID NO. 01. The recombinant bacteria comprise recombinant yeast WAT11 or recombinant agrobacterium GV3101 and the like containing recombinant plasmids. The ring-splitting loganin synthase produced and expressed by the recombinant yeast WAT11 can be used for catalyzing loganin acid to generate sweroside, gentiopicroside or ring-splitting loganin acid. The split-ring loganin synthase produced and expressed by the recombinant agrobacterium GV3101 catalyzes loganin acid in tobacco leaves to produce gentiopicroside.

The present disclosure concerns recombinant yeast host cells having a first genetic modification for downregulating a first metabolic pathway that converts NADP+ to NADPH, as well as a second genetic modification for upregulating a second metabolic pathway that converts NADP+ to NADPH. The second genetic modification allows the expression of a glyceraldehyde-3-phosphate dehydrogenase lacking phosphorylating activity, which can, in some embodiments, be from enzyme commission 1.2.1.9 or 1.2.1.90. The second pathway is distinct from the first metabolic pathway. The present disclosure also concerns a process for making and improving the yield of a fermented product, such as ethanol, using the recombinant yeast host cell.

This invention discloses a recombinant yeast for producing α-ionone, its construction method, and its applications. The construction method involves using a yeast encoding osmanthus flowers... Osmanthus fragrans The gene for the carotenoid cleavage dioxygenase OfCCD1 was obtained through rational modification and codon optimization. OfCCD1m The gene, with its 5' end fused to the N-terminal membrane-anchored peptide coding sequence of human tyrosine kinase LCK, was transformed into the δ site of Saccharomyces cerevisiae TW3 to obtain recombinant yeast TW4 producing α-ionone. The Saccharomyces cerevisiae strain TW4 producing α-ionone of this invention has the following beneficial effects: it achieves de novo synthesis of α-ionone in Saccharomyces cerevisiae for the first time, filling a technological gap in this field; by using a membrane anchoring strategy to locate the OfCCD1m enzyme in the yeast plasma membrane, the contact efficiency between the enzyme and the substrate ε-carotene is significantly improved, resulting in an α-ionone yield of 1.46 mg / L.

Disclosed are a novel nuclear localization sequence (NLS) mutant and a method for improving biosynthetic efficiency using same, belonging to the field of bioengineering. According to the present disclosure, a novel NLS is screened out, and recombinant yeast with improved metabolite synthesis efficiency is constructed by applying the NLS obtained through screening, which provides key technical support for developing a high-yield, stable and cost-effective yeast biosynthesis platform, and facilitates the use of recombinant yeast as a cell factory for large-scale industrial biomanufacturing.

The invention provides a recombinant yeast expression system of trypsin free from enterokinase activation, a construction method and application of the recombinant yeast expression system, modified trypsinogen coding nucleic acid, a recombinant yeast host cell, active trypsin and a production system. The propeptide sequence is connected with a mature enzyme sequence; and a mature enzyme sequence, wherein a basic amino acid residue is contained between the propeptide sequence and the mature enzyme sequence. According to the technical scheme, the problem that in the prior art, recombinant trypsin production must depend on exogenous enterokinase for activation can be effectively solved.