43 results about "Yeast genome" patented technology

The invention belongs to the technical field of bioengineering, and discloses a recombinant yarrowia lipolytica capable of efficiently and rapidly producing sclareol, which is characterized in that optimized sclareol synthase related coding genes ScLPPS, ScSCS, GGPPS1, GGPPS2, ERG20 and tHMGR coding genes are inserted into a yarrowia lipolytica genome; inserting optimized pentadienol utilization pathway coding genes such as IPK, THIM and IDI coding genes; and inserting an optimized ATP: citrate lyase coding gene ACLY to obtain the ATP: citrate lyase coding gene ACLY. The recombinant yarrowia lipolytica is based on knockout of a yarrowia lipolytica strain responsible for coding a non-homologous recombination gene ku70, so that the homologous recombination capability of the yarrowia lipolytica strain is enhanced, gene integration is realized through the homologous recombination function of the yarrowia lipolytica, the genetic stability of the introduced gene can be greatly improved, and the recombinant yarrowia lipolytica strain has a good application prospect. The construction method of the recombinant yarrowia lipolytica strain is efficient and simple to operate.

The invention relates to a high-activity recombinant XVII type collagen as well as a rapid preparation method and application thereof, and belongs to the technical field of bioengineering. The amino acid sequence of the high-activity recombinant XVII type collagen is as shown in SEQ ID NO. 1, and the nucleotide sequence of the high-activity recombinant XVII type collagen is as shown in SEQ ID NO. 2. The invention also provides a preparation method and application of the recombinant XVII type collagen. The high-activity recombinant XVII type collagen has good biological activity, has good effects of promoting cell adhesion, cell proliferation and cell migration, and can be compounded with other proteins to form raw materials of tissue engineering products, cosmetics, health care products or medicines. The high-activity recombinant XVII type collagen gene is integrated to different positions of a yeast genome through three different vectors, and the copy number of a target gene is increased, so that the expression quantity of the recombinant XVII type collagen is remarkably increased, and the yield of the high-activity recombinant XVII type collagen is effectively increased.

The invention discloses a Sarcabomyces lipolytica strain for synthesizing (2S)-naringenin and application of the Sarcabomyces lipolytica strain, and belongs to the technical field of engineering and bioengineering. According to the invention, a key enzyme CHS is subjected to semi-rational transformation, so that the yield of the strain (2S)-naringenin expressing the mutant V196A / T132C is increased by 60%. According to the method, the modified CHS is further integrated into a yeast genome, conversion of p-coumaric acid to (2S)-naringenin is promoted by strengthening a shikimic acid pathway and enhancing supply of cofactors, so that the yield of the (2S)-naringenin reaches 776mg / L after the constructed strain is fermented in a 24-pore plate for 96 hours, and the yield of the constructed strain can reach 8.65 g / L after the strain is fermented in a 5L fermentation tank; the method lays a foundation for the subsequent biosynthesis of flavonoid compounds, and has potential value and significance for the development of synthetic biology.

The invention belongs to the field of biosynthesis, and in particular relates to LBD (Levandanenediol Pyrophosphate) synthase and engineered yeast for producing sclareol. In order to solve the problem that in the prior art, the sclareol biosynthesis yield is generally low, LSLPPs mutant containing at least one mutation of D730L, D374L, N674L or G379M is obtained by performing mutation and optimization on LSLPPs pyrophosphate synthase, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability caused by free plasmid expression of part of saccharomyces cerevisiae strains, a sclareol synthesis pathway is constructed in a yeast genome, and meanwhile, an acetyl coenzyme A synthesis pathway, an MVA pathway and a sclareol synthesis pathway are enhanced; chassis bacteria for synthesizing sclareol are modified in a manner of knocking out part of transcription inhibition factors, and the yield of a 5L fermentation tank reaches 26.11 g / L and is greatly improved compared with the prior art.

The invention belongs to the field of biosynthesis, and particularly relates to sclareol synthase and engineered yeast for producing sclareol. In order to solve the problem that in the prior art, sclareol biosynthesis yield is generally low, sclareol synthase is mutated and optimized, an SsScs mutant containing at least one mutation of Y491F, G307L or N269F is obtained, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability caused by free plasmid expression of part of saccharomyces cerevisiae strains, a sclareol synthesis pathway is constructed in a yeast genome, and meanwhile, an acetyl coenzyme A synthesis pathway, an MVA pathway and a sclareol synthesis pathway are enhanced; chassis bacteria for synthesizing sclareol are modified in a manner of knocking out part of transcription inhibition factors, and the yield of a 5L fermentation tank reaches 21.13 g / L and is greatly improved compared with the prior art.

The invention discloses a yeast engineering bacterium for producing apigenin and application thereof. The yeast engineering bacterium comprises a yeast genome, a whole-cell catalytic synthesis apigenin module, a heterologous apigenin biosynthesis path and an NADPH supply module, the whole-cell catalytic synthesis apigenin module comprises an FNSI coding gene and an FNSII coding gene; the heterologous apigenin biosynthesis path comprises a TAL coding gene, a 4CL coding gene, a CHS coding gene, a CHI coding gene and an FNSII coding gene; the NADPH supply module comprises a G6PD coding gene and a PGD coding gene; the method disclosed by the invention has the characteristics of greenness and safety, and lays a foundation for synthesizing apigenin in a yeast cell factory.

The invention belongs to the technical field of bioengineering, and particularly relates to kluyveromyces marxianus engineering bacteria for synthesizing eicosapentaenoic acid and application of the kluyveromyces marxianus engineering bacteria. According to the engineering bacterium, pyruvate decarboxylase PDC1 genes, alcohol dehydrogenase ADH1 genes and alcohol dehydrogenase ADH2 genes are sequentially knocked out from a kluyveromyces marxianus FIM1 delta ura3 strain; meanwhile, a citrate lyase ACL gene, a pyruvate oxidase PO gene and a phosphate transacetylase PTA gene are inserted into the loci respectively, so that synthesis of acetyl coenzyme A is enhanced; then a plurality of genes are sequentially inserted into a Kluyveromyces marxianus genome, and then a vector of a delta 8 desaturase delta 8D gene is transferred into the Kluyveromyces marxianus genome. The invention also provides a method for efficiently preparing the EPA, namely the Kluyveromyces marxianus engineering strain is used as a fermentation strain, glucose, a yeast extract, peptone and other components are used as a culture medium, and the yield of the EPA produced by culturing and fermenting reaches 8.71 mg / g of dry cell weight.

Provided are a brewer's yeast having low ethanol synthesis and high acetyl-CoA synthesis, and a use thereof, belonging to the technical field of biology. In order to reduce the accumulation of ethanol, the main byproduct of brewer's yeast, and increase synthesis of acetyl-CoA, MTH1, MED2 and HXT2 transcription factors are modified, an optimal transcription factor combination for reducing ethanol synthesis is obtained by screening, and then heterologous acetyl-CoA synthesis pathways of different synthesis pathways integrated in the brewer's yeast genome are compared, and a recombinant brewer's yeast strain is provided. The strain can grow in a sterile culture medium using glucose as a carbon source, creating a platform for modifying brewer's yeast by metabolic engineering, to synthesize high-value compounds; in addition, the construction method is simple and easy to use, and has good application prospects.

The invention discloses pichia pastoris for efficiently synthesizing lactoferrin as well as a construction method and application of the pichia pastoris, and belongs to the technical field of biosynthesis. The pichia pastoris for efficiently synthesizing the lactoferrin expresses at least one of KAR2, PDI1 and ERO1 in molecular chaperones in a host in an enhanced manner and carries an optimized secretory signal peptide, and the secretory signal peptide is obtained by replacing a leading peptide of alpha-factor with any signal peptide in LacZ, PelB, FLO and alpha-factor. The yield of LF can be effectively increased by enhancing expression of at least one of KAR2, PDI1 and ERO1 molecular chaperones in molecular chaperones in a host; when the lactoferrin expression cassette is integrated into the recombinant pichia pastoris X-33 genome and the copy number is increased to 2-6, the LF yield can be further increased, and when the copy number is 4, the LF yield reaches a peak value.

The invention discloses an adenosine deaminase mutant. The adenosine deaminase mutant is a C141R mutant and an L144P mutant based on a wild type. The invention also discloses a pichia pastoris base editor containing the adenosine deaminase mutant, the pichia pastoris base editor can realize accurate editing of specific base sites of a pichia pastoris genome, and a gene editing tool is provided for basic and application research based on pichia pastoris.

The invention discloses a method for improving yeast CRISPR-Cas gene editing efficiency, and belongs to the technical field of gene engineering. Firstly, SPC plasmids are transferred into host bacteria, the SPC plasmids contain gRNA1 expression cassettes targeting target genes 1, 5'ends of the gRNA1 expression cassettes are connected with Checkpoint promoters, the gRNA1 expression cassettes serving as host cells are transferred into pCasT plasmids, the plasmids and gRNA2 targeting yeast genome target genes 2 achieve DNA double-strand breakage through Cas protein, the Checkpoint promoters of the SPC plasmids are activated, transcription of the gRNA1 is started, and gene editing of the target genes 1 is achieved. On the contrary, the Cas protein and the gRNA1 expression cassette do not realize gene editing on the target gene 1, the unedited strain dies by absorbing the externally added toxic substances, and the successfully edited strain survives due to the lack of the functions. According to the method, the gene editing efficiency of a single target site can be improved, the gene knockout, large-fragment DNA insertion and recombination efficiency can be improved, and the high-throughput genome editing efficiency is improved.

The present invention relates to an engineered Yarrowia lipolytica bacteria that secretes a large amount of L-astaxanthin, characterized in that the wild-type Yarrowia lipolytica is used as a starting strain, and four gene clusters GGS1-CarB-CarRP, PsCrtW-HpCrtZ, GGS1-CrtYB-CrtI and ACC1-DGA1 are randomly integrated into the wild-type Yarrowia lipolytica genome, and the endogenous gene SNQ2 of Yarrowia lipolytica is overexpressed and the MHY1 gene is knocked out. The engineered bacteria is named Yarrowia lipolytica YYYE112, the deposit number is CGMCC NO.30371, and the deposit date is April 17, 2024. The engineered bacteria provided by the present invention can be fermented with vegetable oil as a carbon source, and have the advantages of fast cell growth rate, high astaxanthin yield and high secretion capacity, which is conducive to large-scale production of astaxanthin and reduces the production and extraction costs of astaxanthin.

The present invention discloses an adenosine deaminase mutant, which is a C141R and L144P mutant based on the wild type. The present invention also discloses a Pichia pastoris base editor containing the adenosine deaminase mutant. The Pichia pastoris base editor can achieve precise editing of specific base sites in the Pichia pastoris genome, providing a gene editing tool for basic and applied research based on Pichia pastoris.

The invention relates to the technical field of bioengineering, and discloses a yeast engineering bacterium for producing squalene and application thereof, the yeast engineering bacterium comprises yeast chassis cells, a heterologous squalene biosynthesis module, an NADPH supply module, a precursor supply module and a grease body engineering module, the specific technological process comprises the following steps: integrating a key enzyme coding gene in a heterologous squalene biosynthesis module into a yeast chassis cell to obtain a yeast genome containing a squalene biosynthesis pathway; integrating the NADPH supply module, the precursor supply module and the grease body engineering module into a yeast genome to obtain the yeast engineering bacteria for producing squalene; and fermenting the yeast engineering bacteria by taking a YYG culture medium as a substrate to obtain squalene. The yeast engineering bacterium constructs and optimizes a biosynthesis integration module required by squalene, realizes adaptation of a squalene biosynthesis pathway and yeast chassis cells, can efficiently and stably produce squalene, and also has the characteristics of greenness and safety.

The invention relates to the technical field of molecular biology, in particular to microsatellite primers based on hansenula polymorpha yeast genomes and application of the microsatellite primers. The invention provides six groups of brand-new microsatellite site primer groups related to hansenula polymorpha. Wherein three groups are cactus hansenula polymorpha primer group sequences, and a cactus hansenula polymorpha yeast region can be divided into four genotypes, namely Hopa, Hopb, Hopc and Hopd, according to the types of electrophoretic bands of PCR (Polymerase Chain Reaction) products at three microsatellite loci HOP1, HOP2 and HOP5. The three groups are rare hansenula polymorpha primer group sequences, and the rare hansenula polymorpha can be divided into four genotypes, namely Hoca, Hocb, Hocc and Hocd, according to the types of electrophoretic bands of PCR (Polymerase Chain Reaction) products at three microsatellite sites, namely HOC2, HOC4 and HOC7.

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.

The present invention discloses flavonoid 4'-O-methyltransferase and its application in the production of hesperetin, belonging to the field of genetic engineering and bioengineering technology. The present invention screened and identified new flavonoid 4'-O-methyltransferases derived from tea-branch orange and Huazhou pomelo; the flavonoid 4'-O-methyltransferases CrcOMT-2 and CgtOMT-3 obtained by screening were integrated and expressed in the multi-copy site rDNA and ZETA site on the genome of Yarrowia lipolytica YE26, respectively, to achieve the de novo synthesis of hesperetin. The Yarrowia lipolytica Z02 constructed by the present invention achieved the de novo synthesis of 130.2 mg / L hesperetin in a 5L fermentor, which is also the highest yield of hesperetin de novo synthesis in microorganisms reported, laying the foundation for the subsequent biosynthesis of flavonoids.

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 application discloses a Yarrowia lipolytica strain for synthesizing (2S)-naringenin and application thereof, and belongs to the field of genetic engineering and bioengineering technology. The application improves the yield of (2S)-naringenin by 60% through semi-rational modification of a key enzyme CHS, and further integrates the modified CHS into the yeast genome. In addition, the application promotes the conversion of coumaric acid to (2S)-naringenin by strengthening the shikimic acid pathway and enhancing the supply of auxiliary factors, so that the yield of (2S)-naringenin reaches 776 mg / L after 96 hours of fermentation of the constructed strain in a 24-hole plate, and the yield reaches 8.65 g / L in a 5L fermenter. The application lays a foundation for subsequent biosynthesis of flavonoids, and has potential value and significance for the development of synthetic biology.

This invention belongs to the field of biosynthesis, specifically relating to a variant of lysine pyrophosphate diol synthase and the constructed engineered yeast strain for perillaldehyde synthesis. To address the generally low yield of perillaldehyde biosynthesis in existing technologies, this invention mutates and optimizes the lysine pyrophosphate diol synthase to obtain an SsLPPs mutant containing the G379M mutation, which is then used for perillaldehyde synthesis. Simultaneously, to address the genetic instability issues associated with free plasmid expression in some Saccharomyces cerevisiae strains, this invention constructs a perillaldehyde synthesis pathway within the yeast genome. Furthermore, by enhancing the acetyl-CoA synthesis pathway, the MVA pathway, and the perillaldehyde synthesis pathway, and by knocking out some transcriptional repressors, the substrate strain synthesizing perillaldehyde is modified, resulting in a significantly increased yield compared to existing technologies.

The application discloses a kind of production cordycepin yeast engineering bacteria and application.The yeast engineering bacteria is inserted in the genome of yeast cordycepin biosynthesis module, and purine metabolic pathway overexpression module or ATP and precursor supply module;The cordycepin biosynthesis module is composed of Cns1 gene and Cns2 gene;The purine metabolic pathway overexpression module is composed of ADE4 gene and Cns3-NK gene;The ATP and precursor supply module is composed of PGK gene and Cns3-NK gene.The application is according to the biosynthesis pathway of cordycepin and its metabolic regulation, constructs and optimizes the biosynthesis integrated module required in yeast in vivo cordycepin, creates yeast cell factory, and establishes microbial fermentation production cordycepin technical system with cheap carbon source sugarcane molasses as raw material, realizes the efficient, green, safe production of cordycepin.

The invention belongs to the field of gene engineering and enzyme engineering, and relates to lipase, a coding gene Lip1.1, a genetically engineered bacterium of the lipase and application of the lipase and the coding gene Lip1.1. The invention aims to provide lipase with a specific nucleotide sequence (SEQ ID NO. 1), and further provides a lipase gene, an extracellular secretion expression engineering bacterium and application of the lipase gene in the field of foods or medicines. Particularly, the invention emphasizes the application potential of the lipase gene in the aspect of fish oil fatty acid. The invention provides a lipase gene sequence which is integrated into a yeast genome through a plasmid vector, so that the high expression stability and the exocytosis capacity of target encoding protein are ensured. The recombined target gene fragment can be transferred to any strain with strong lipase production capacity, and the strong expandability of the recombined target gene fragment is embodied. The characteristic not only enhances the utilization efficiency of gene resources, but also provides powerful support for popularization of lipase in various industrial applications.

The invention discloses a preparation method of a protease overexpression yeast strain, the protease overexpression yeast strain and application of the protease overexpression yeast strain, and relates to the technical field of molecular biology.According to the technical scheme, a first expression plasmid of protease A is provided; carrying out one-step assembly on the first expression plasmid, a second expression plasmid containing the TPI1 gene and a third basic vector, and screening and verifying to obtain a third expression plasmid; the third expression plasmid is transferred into a yeast strain, a protease overexpression yeast strain is obtained through screening and verification, the yeast strain is a TPI1 gene knockout strain, a TPI1 gene transcription unit and a protease A gene transcription unit are connected in series and introduced into the yeast strain with the TPI1 gene knockout in a plasmid form, and overexpression of protease is achieved. The yeast genome does not need to be directly edited. The method not only simplifies the construction process, but also possibly improves the genetic stability of the engineering strain, and reduces the potential risk caused by genome integration.

Provided is a method for constructing an engineered yeast for de novo synthesis of mogroside. The method comprises: inserting constructed cucurbitadienol synthase gene expression cassette Ptef1-cds-Tcyc1, epoxide hydrolase gene expression cassette Ptef1-eph3-Tcyc1, cytochrome P450 enzyme-encoding gene CYP and cytochrome P450 enzyme reductase gene expression cassette Ttdh3-cyp87d18-P gal1 / 10-atcpr2-Tcyc1, and glycosyltransferase-encoding gene expression cassette Tadh1-ugt74ac1-Pgal1 / 10-ugtms1-Tcyc1 into a yeast genome, so as to obtain an engineered yeast for mogroside production. The engineered yeast can be used for de novo synthesis of mogroside by using glucose as a substrate for fermentation.

The invention belongs to the field of biosynthesis, and in particular relates to an improved variant of libdanenediol pyrophosphate synthase and engineered yeast for producing sclareol. In order to solve the problem that in the prior art, the sclareol biosynthesis yield is generally low, LsLPPs mutant containing N674L mutation is obtained by performing mutation and optimization on LsLPPs pyrophosphate synthase, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability caused by free plasmid expression of part of saccharomyces cerevisiae strains, a sclareol synthesis pathway is constructed in a yeast genome, and meanwhile, an acetyl coenzyme A synthesis pathway, an MVA pathway and a sclareol synthesis pathway are enhanced; and the chassis bacteria for synthesizing sclareol are modified in a manner of knocking out part of transcription inhibition factors, so that the yield is greatly improved compared with the prior art.

The invention discloses a saccharomyces cerevisiae genetically engineered bacterium for efficiently synthesizing mogroside V through multi-dimensional cytoplasm adaptability and a construction method of the saccharomyces cerevisiae genetically engineered bacterium. The construction method of the saccharomyces cerevisiae genetically engineered bacterium comprises the following steps: integrating truncated 3-hydroxy-3-methylglutaryl coenzyme A reductase tHMG1, pentenyl pyrophosphate isomerase IDI1, squalene synthetase ERG9, epoxy squalene cyclization enzyme ERG1, choline kinase ScCK, isopentenyl phosphokinase AtIPK, phosphoketolase BbPK, phosphotransacetylase PTA and dihydroxyacetone hydrolase EcHAD at a site of a saccharomyces cerevisiae genome GAL80, and carrying out enzymolysis, so as to obtain the saccharomyces cerevisiae genetically engineered bacterium. The invention relates to a preparation method of a cytochrome P450 enzyme, which comprises the following steps of: preparing a cycloepoxide hydrolase SgEPH3, a cytochrome P450 enzyme CYP87D18 and a cytochrome P450 enzyme reductase AtCPR1; an ABC efflux protein PDR11, a glycosyl transferase UGTMG1, a sucrose synthase Susy and a glycosyl transferase SgUGT94-289-3 are integrated at a site Exg1 of a saccharomyces cerevisiae genome. The saccharomyces cerevisiae genetically engineered bacterium constructed by the invention realizes a higher level of mogroside V yield synthesized from the beginning in microorganisms, and has important industrial application potential.

The invention relates to the technical field of saccharomyces cerevisiae genetic engineering transformation, in particular to a high-throughput saccharomyces cerevisiae transformation method. The method comprises the following steps: screening an IGR sequence on a saccharomyces cerevisiae genome through bioinformatics, respectively taking 45bp from upstream and downstream of the IGR sequence as genome homologous sequences for primer design, designing and subpackaging a PCR premixed solution and the designed primer by using an automatic platform script for PCR amplification, adding an obtained PCR product into competent cells, adding a conversion solution, and carrying out PCR amplification on the competent cells to obtain the saccharomyces cerevisiae gene. Performing heat shock; washing and resuspending the cells subjected to heat shock, and then coating the cells on a screened solid culture medium for culturing; the conversion process is carried out on an automatic platform island. According to the method, the transformation efficiency is improved, the operation time of high-throughput transformation is shortened, the intelligence and stability of each transformation are improved, and the method is suitable for large-scale yeast genome editing, functional gene screening and saccharomyces cerevisiae genome site screening.

The present application belongs to the field of biosynthesis, and particularly relates to a sclareol synthase and a yeast engineering bacterium for producing sclareol. In order to solve the problem of low yield of sclareol biosynthesis in the prior art, the sclareol synthase is mutated and optimized in the present application, a SsScs mutant containing at least one mutation of Y491F, G307L or N269F is obtained, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability existing in the expression of free plasmid in some Saccharomyces cerevisiae strains, the present application constructs a sclareol synthesis pathway in the yeast genome, strengthens the acetyl coenzyme A synthesis pathway, the MVA pathway and the sclareol synthesis pathway, and knocks out part of the transcriptional repressor to modify the chassis bacterium for synthesizing sclareol. The yield in a 5 L fermenter reaches 21.13 g / L g / L, which is greatly improved compared with the prior art.

The invention discloses a qPCR (quantitative polymerase chain reaction) method for detecting saccharomyces cerevisiae, Wickerhamomyces anomalus and zygosaccharomyces bayer and application of the qPCR method, and belongs to the technical field of bioengineering. The invention establishes a method for screening specific gene segments and designing qPCR primers based on multi-yeast genome comparison, successfully identifies the specific genes of saccharomyces cerevisiae, Wickerhamomyces anomalus and zygosaccharomyces bayer, verifies high specificity in non-target yeast, establishes qPCR standard curves of the three yeasts, and provides a basis for screening the specific genes of the saccharomyces cerevisiae, the Wickerhamomyces anomalus and the zygosaccharomyces bayer. The linear range is 103-108 copies / [mu] L, the detection limit reaches 10 copies / [mu] L, and the quantification limit reaches 103 copies / [mu] L. The method realizes quantitative and qualitative detection of saccharomyces cerevisiae, Wickerhamomyces anomalus and zygosaccharomyces bayer in a pure bacterium sample and a simulated fermentation sample, has the characteristics of high sensitivity, high specificity, high stability and the like, and is accurate and reliable in detection result.

The invention discloses a method for rapidly and quantitatively detecting the number of saccharomycetes cells in honey. The method comprises the following steps: (1) separating the saccharomycetes cells in the honey; (2) extracting a yeast genome; (3) carrying out real-time fluorescent PCR (Polymerase Chain Reaction) (Taqman qPCR) amplification; and (4) constructing a standard curve. According to the method, accurate quantification of the total number of saccharomycetes in the honey is realized through a specific molecular detection technology, and reliable technical support is provided for quality control, safety evaluation and decay early warning of honey products. The method successfully provides a first specific quantitative solution for yeast pollution in honey, and has high sensitivity, rapidity and universality, so that the method has important application value in the aspects of honey production quality monitoring, storage and transportation process risk early warning and food safety standard upgrading.