77 results about "Kluyveromyces sp." patented technology

The invention relates to a Kluyveromyces cerevisiae gene modified cell containing GDP-mannose dehydratase polypeptide, which can be applied to synthesis of human milk oligosaccharide, and belongs to the field of synthetic biology. According to the technical scheme, the gene modified cell comprises a recombinant nucleic acid sequence for coding a polypeptide with GDP-mannose dehydratase polypeptide (GDP-mannose dehydratase polypeptide) activity. The polypeptide capable of catalyzing the conversion of GDP-mannose into GDP-4-keto-6-deoxymannose comprises a polypeptide of which the amino acid sequence is at least 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical to the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2 and / or the amino acid sequence of polypeptides M1-M13, wherein the amino acid sequence of the polypeptide is at least 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical to the amino acid sequence of the polypeptides M1-M13. The invention provides a kluyveromyces kluyveromyces genetically modified cell with fucosyllactose synthesis activity.

The invention belongs to the technical field of biology, and discloses Kluyveromyces lactis FRI and application thereof, the preservation number of the Kluyveromyces lactis FRI is CGMCC (China General Microbiological Culture Collection Center) No.30910, and the Latin name of the Kluyveromyces lactis FRI is Kluyveromyces lactis. The preservation date is June 24, 2024, and the preservation unit is China General Microbiological Culture Collection Center (CGMCC). The invention provides a Kluyveromyces lactis FRI, which has very high lignin degrading enzyme activity. The Kluyveromyces lactis FRI can grow in straws and has strong utilization capability on the straws.

The invention belongs to the field of fermentation, and particularly relates to kluyveromyces marxianus, a complex microbial inoculant, fermented tea primary pulp as well as a preparation method and application of the kluyveromyces marxianus. Wherein the Kluyveromyces marxianus is Kluyveromyces marxianus Linzhi.KM-D3, the Kluyveromyces marxianus Linzhi.KM-D3 is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number of the Kluyveromyces marxianus Linzhi.KM-D3 is CCTCC NO: M 20231575. The DPPH free radical scavenging capacity of the fermented tea raw stock obtained through fermentation of the kluyveromyces marxianus is larger than or equal to 140%, the hydroxyl free radical scavenging capacity is larger than 130%, the ABTS free radical scavenging capacity is larger than 80%, and the SOD enzyme activity of each L of the fermented tea raw stock is larger than 1300U. Meanwhile, the pH value of the fermented tea puree provided by the invention is less than or equal to 4.0, the total acid content (in terms of lactic acid) is greater than or equal to 6g / 100g, the ethanol content is less than or equal to 0.5 g / 100g, the sucrose content is less than or equal to 0.5 g / 100g, and the fermented tea puree has no fermentation peculiar smell and light bitter taste in the aspect of sensory evaluation.

The invention provides a genetically engineered bacterium for producing polydeoxyribonucleotide as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium comprises kluyveromyces marxianus for expressing a salmon DNA polymerase I gene. According to the invention, the edible probiotic Kluyveromyces marxianus is transformed through genetic engineering, efficient, safe and controllable production of PDRN is realized, the biological activity of PDRN is equivalent to that of PDRN from a natural source, and large-scale production and application of PDRN are facilitated.

The invention relates to the field of microbial food fermentation, in particular to kluyveromyces marxianus capable of producing high-yield mellow substances and application of the kluyveromyces marxianus. The Kluyveromyces marxianus capable of producing the high-yield mellow substances is obtained through screening, the strain is named as Kluyveromyces marxianus Y4, the preservation number is CCTCC (China Center for Type Culture Collection) NO: M2025759, the preservation date is April 11, 2025, and the preservation address is China Center for Type Culture Collection. The Kluyveromyces marxianus obtained by the invention is applied to the preparation of the mellow flavored yoghourt, and has the advantages that the yield of the mellow substances is increased; the variety and the content of alcohol volatile flavor substances in the yoghourt can be remarkably improved, and richer taste levels and unique flavor are provided for the yoghourt.

The invention relates to the technical field of environmental microorganisms and resource fermentation, and discloses a Kluyveromyces marxianus BMK7 for protein production and biogas slurry purification and application of the Kluyveromyces marxianus BMK7, the BMK7 is Kluyveromyces marxianus and is obtained through ultraviolet mutagenesis and hygromycin B adaptive evolution, the preservation number is CCTCC NO: M 20251050, and the Kluyveromyces marxianus BMK7 has good growth performance and genetic stability. The strain can be efficiently fermented under the condition that biogas slurry serves as a main substrate, ammonia nitrogen and chemical oxygen demand (COD) in the biogas slurry are remarkably reduced, and meanwhile high-proportion mycoprotein is synthesized. The thalli have strong flocculability and good sedimentation characteristics, and are beneficial to protein recovery. The method can be widely applied to treatment and resource utilization of sewage such as breeding wastewater, kitchen fermentation liquor and anaerobic effluent, and has a good industrialization prospect.

The invention belongs to the technical field of microorganisms, and particularly relates to kluyveromyces marxianus and application thereof in preparation of rose fruity fermented milk. The Kluyveromyces marxianus strain is named as ALE-X20, is preserved in the China General Microbiological Culture Collection Center (CGMCC), is classified and named as Kluyveromyces marxianus, has the preservation number of CGMCC NO.32221, and is preserved on October 15, 2024. The Kluyveromyces marxianus ALE-X20 disclosed by the invention can efficiently convert an amino acid substrate into a key flavor substance, namely, 2-phenethyl alcohol, of rose fruit fragrance, so that the unique rose fragrance is given to the fermented milk under the condition that no essence or spice is added.

Disclosed herein are genetically engineered Kluyveromyces sp. cells capable of producing D-allulose. The genetically engineered Kluyveromyces sp. cells comprise an exogenous polynucleotide sequence encoding an allulose-6-phosphate 3-epimerase enzyme at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or100% identical to at least one of SEQ ID NOs:249-256, 258, and 259; and an exogenous polynucleotide sequence encoding an allulose-6-phosphate phosphatase enzyme at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identical to at least one of SEQ ID NOs:87, 89, 190, 123, 105, 107, 115, 83, 95, 113, 117, 119, 121, 127, 131, 137, 145, 169, 173, 179, and 183.

The invention discloses fermented soybean meal and a preparation method thereof, and belongs to the technical field of fermented food. After soybean meal and soybean slurry are mixed, bacillus natto and kluyveromyces marxianus are inoculated for fermentation, and the fermented soybean meal can be obtained. According to the method, bacillus natto and kluyveromyces marxianus are used as fermentation strains, a soybean meal processing technology is optimized through a single-bacterium and mixed-bacterium fermentation technology, the influence of fermentation on physicochemical properties, functional activity and flavor quality of soybean meal is systematically explored, a high-value fermented soybean meal product is prepared, and the nattokinase (NK) activity of the mixed-bacterium fermented soybean meal reaches 6562.49 IU / g. High-activity nattokinase can be produced by fermenting soybean meal with bacillus natto, and the flavor of the fermented soybean meal can be improved and the nutritional value of the soybean meal can be improved by mixed fermentation with kluyveromyces cerevisiae. According to the technical scheme, high-value utilization of soybean meal resources can be promoted, the cost of terminal products can be reduced, and dependence on soybean import can be reduced.

The present invention belongs to the technical field of microbial fermentation, and particularly relates to a Spirulina probiotic fermentation composition for preventing and treating metabolic associated fatty liver disease, a preparation method and an application. The present invention uses a combination of Lactobacillus acidophilus and Kluyveromyces marxianus to ferment Spirulina to obtain a fermentation composition for preventing and treating metabolic associated fatty liver disease; the Lactobacillus acidophilus was deposited at the China Center for Type Culture Collection on August 29, 2016, with the deposit address: Wuhan University, Wuhan, China, and the taxonomic name: Lactobacillus acidophilus KDB-03 ( Lactobacillus acidophilus KDB-03), deposit number: CCTCC NO: M 2016429. This fermentation composition can significantly increase the total flavonoid and total phenolic contents of Spirulina, enhance its antioxidant activity; significantly enhance the amino acid level in the liver of mice induced by a high-fat diet, significantly improve the metabolic profile of liver tissue, and relieve metabolic associated fatty liver disease induced by a high-fat diet; and can also effectively remove the fishy smell of Spirulina itself.

The invention belongs to the technical field of biology, and particularly relates to organic zinc-rich kluyveromyces marxianus and application of the organic zinc-rich kluyveromyces marxianus in preparation of quick-frozen rice and flour products, the zinc content of the organic zinc-rich kluyveromyces marxianus is 1000-15000 mg / kg, and the organic zinc proportion is not lower than 90%. And by adding Kluyveromyces marxianus containing high-concentration organic zinc, the content of the organic zinc in the product can be increased, the frost crack phenomenon of the quick-frozen rice and flour product in the cryopreservation process can be effectively avoided, the taste of the frozen rice and flour product can be remarkably improved, the nutrient elements of the frozen rice and flour product are enriched, and the growth and development of the body are promoted.

The invention discloses a method for enhancing the nutritional value of microbial mycoprotein. Protein rich in essential amino acid is heterologously expressed in the recombinant kluyveromyces marxianus, and the contents of crude protein, essential amino acid required for feeding human bodies and animals and branched chain amino acid in yeast obtained by fermentation are increased to different extents, so that the nutritional value of mycoprotein is enhanced. The method can be used for producing high-quality microbial protein, and has wide application value in the fields of food and feed.

The present application relates to a kind of preparation methods of legume fermented food, comprising the following steps: (1) providing first soybean milk, access first fermentation bacteria, anaerobic fermentation, fermentation end point is pH 4-5, obtain first fermentation product;The first fermentation bacteria are selected from one or more lactic acid bacteria;(2) providing second soybean milk, access second fermentation bacteria, anaerobic fermentation, fermentation length is 6-10h, obtain second fermentation product;The second fermentation bacteria are selected from one or more of Leuconostoc mesenteroides, Kluyveromyces marxianus and Saccharomyces cerevisiae;(3) according to 0.5-1.5:1 Weight ratio mixes first fermentation product with second fermentation product, aerobic fermentation is carried out to mixture, fermentation end point is to pH 4-6, obtain third fermentation product.

The application provides a method for improving the LPS stress resistance of broilers, which is realized by feeding a mannose oligosaccharide compound preparation prepared from enzymolysis coffee grounds. The preparation method of the mannose oligosaccharide compound preparation comprises the following steps: crushing coffee grounds, and adding a compound reagent for pretreatment; adding a compound enzyme preparation I to the mixed system after reaction, and carrying out enzymolysis for 6-8 hours under the condition of 50-60 DEG C; then adding a compound enzyme preparation II, and carrying out sufficient enzymolysis under the condition of 55-65 DEG C; after the enzymolysis is completed, a fermentation compound microbial agent is used for continuing fermentation for 6-12 hours under the condition of 30-35 DEG C, and drying is carried out; the compound enzyme preparation I comprises papain, alpha-amylase and saccharifying enzyme; the compound enzyme preparation II comprises cellulase, mannanase and glucanase; and the fermentation compound microbial agent comprises maxwell kluyver yeast and lactobacillus plantarum. The method can not only avoid drug residues caused by feeding drugs, but also realize the reuse of coffee grounds.

This invention provides a feed additive and compound feed for preventing and treating coccidiosis in broilers, comprising the following components: *Kluyveromyces martensii*, ferulic acid esterase, aminopeptidase, and glucanase. Preferably, the mass percentage of *Kluyveromyces martensii* is 30-40%; the mass percentage of ferulic acid esterase is 15-28%; the mass percentage of aminopeptidase is 15-25%; and the mass percentage of glucanase is 18-25%. Based on extensive research, this invention has determined that this combination has a significant effect on coccidiosis control in broilers, effectively improving broiler production performance indicators, increasing economic benefits, and reducing the use of anticoccidial drugs.

The invention discloses a Kluyveromyces marxianus engineering strain and an application thereof in high-temperature fermentation production of 3-hydracrylic acid, which are characterized in that heat-resistant Kluyveromyces marxianus is used as a platform, a gene editing technology is utilized, a glycerin metabolism way of the Kluyveromyces marxianus is reconstructed, and MCRN940V / K1106W / S1114R mutant gene is over-expressed to finally obtain a YZB521 strain. Under the aerobic condition of 37-42 DEG C, 21.26 g / L of 3-hydracrylic acid and 23.95 g / L of 3-hydracrylic acid can be synthesized through glycerol fed-batch fermentation, and 10.31 g / L of 3-hydracrylic acid and 9.02 g / L of 3-hydracrylic acid can be synthesized through industrial waste glycerol fed-batch fermentation.

The invention discloses a construction method and application of a kluyveromyces marxianus mutant, and belongs to the field of synthetic biology and molecular biology. According to the method, single-stranded DNA binding protein and cytosine deaminase pmCDA1 are fused, a random base editing system is constructed, and whole-genome efficient C-T / G-A mutation is realized in kluyveromyces marxianus by taking MCM2-7 as a verification target. The system does not need a specific target or an exogenous DNA template, the mutation efficiency is high, the distribution is wide, the mutation rate is improved by 303.1 times compared with that of a control group through optimal combination, and the application potential of kluyveromyces marxianus in industrial production is expanded. The method is combined with an efficient screening system, mutant strains with the intracellular protein expression quantity remarkably improved are screened out, the constructed mutation system is not only suitable for kluyveromyces marxianus cell factory transformation, but also can be popularized to genome evolution of other yeasts or eukaryotic hosts, and a new strategy is provided for industrial microorganism breeding and protein engineering.

Disclosed herein are genetically engineered Kluyveromyces marxianus cells capable of producing D-allulose. The genetically engineered Kluyveromyces marxianus cells comprise a deletion or disruption in a native PFK1 gene; an exogenous polynucleotide sequence encoding an allulose-6-phosphate 3-epimerase enzyme at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identical to at least one of SEQ ID NOs:192, 194, and 196, and an exogenous polynucleotide sequence encoding an allulose-6-phosphate phosphatase enzyme at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identical to at least one of SEQ ID NOs:87, 89, 190, 123, 105, 107, 115, 83, 95, 113, 117, 119, 121, 127, 131, 137, 145, 169, 173, 179, and 183.

This invention relates to the field of food fermentation technology, specifically to a high-efficiency fermentation process for purple sweet potato rice wine. It aims to solve the technical problems of significant anthocyanin degradation and fading, difficulty in balancing flavor and efficiency, poor strain adaptability, and short shelf life during the fermentation of purple sweet potato rice wine. This invention improves product quality and fermentation efficiency by constructing a specialized compound fermenting agent and employing a two-stage fermentation process with dynamic pH control. The compound fermenting agent consists of *Rhizopus oryzae*, *Monascus purpureus*, *Kluyveromyces martensii*, and *Lactobacillus plantarum*. The fermentation process employs two stages: the first stage is aerobic fermentation, which promotes the stable existence of anthocyanins and facilitates enzyme activity; the second stage is anaerobic fermentation, which further stabilizes the color and inhibits unwanted microorganisms. This invention uses ultra-high pressure sterilization combined with post-treatment with natural antioxidants to sterilize and preserve the purple sweet potato rice wine, which can improve the pigment retention rate and significantly extend the shelf life of the purple sweet potato rice wine.

The invention discloses comprehensive cereal enzyme powder and a preparation method thereof, and belongs to the technical field of functional food processing. The comprehensive cereal enzyme powder provided by the invention is prepared from the following components in parts by mass: 40-60 parts of a cereal group, 10-30 parts of a berry group and 20-35 parts of a rhizome group, the cereal group comprises germinated brown rice, highland barley and chickpeas; the berry group comprises blueberry pomace and red pitaya peel; the rhizome group comprises purple carrots and jerusalem artichoke powder. Through raw material three-dimensional compounding, strain time sequence control (Aspergillus niger-Kluyveromyces marxianus-Lactobacillus acidophilus) and a low-temperature activity protection process, the core problems of single function, poor stability, low process suitability and the like of the existing enzyme product are solved, and the enzyme has a wide application prospect.

The invention discloses an application of Kluyveromyces marxianus metabolite in preparation of drugs for reducing uric acid, protecting kidney and protecting intestinal tracts, and belongs to the technical field of medicines. The invention provides an oral composition / preparation which takes a Kluyveromyces marxianus fermentation metabolite as an effective component, and by inhibiting the activity of a purine metabolism key enzyme, regulating a kidney / intestinal uric acid transporter and remodeling the intestinal microecology, the effects of reducing uric acid and synchronously protecting the kidney and the intestine are realized, and the problems of single target spot, toxic and side effects, limitation on long-term medication and the like of the existing medicine are solved. The metabolite can significantly reduce serum uric acid, can protect kidney and intestinal functions, and realizes multiple effects of reducing uric acid, protecting kidney and protecting intestines. Compared with a single effect of an existing medicine, the kluyveromyces marxianus metabolite shows a more comprehensive treatment effect through regulation of intestinal flora, inhibition of xanthine oxidase and regulation and control of uric acid transporter.

The invention relates to the technical field of minced fillet deodorization, and discloses a minced fillet deodorization process for fish ball production, which comprises the following steps: (1) cleaning and dehydrating a minced fillet raw material, and controlling the water content at 80-95% after the raw material is dehydrated to obtain a primary minced fillet raw material; (2) activating a composite fungus material, adding the activated composite fungus material into the primary minced fillet raw material, uniformly stirring, fermenting at 28 DEG C for 50-60 minutes, heating to 30 DEG C, and fermenting for 50-60 minutes; wherein the composite fungus material comprises rhizomucor miehei, kluyveromyces marxianus, lactococcus lactis and saccharomyces cerevisiae; and (4) heating the minced fillet obtained in the step (3) to 65-68 DEG C, keeping the temperature for 2-3 minutes, rinsing with ice water, and dehydrating to obtain the fishy smell removed minced fillet. According to the method, the effect of removing the fishy smell of the minced fillet can be improved.

The invention provides a preparation method of germinated brown rice. The preparation method mainly comprises the following steps: screening brown rice, soaking the brown rice in a sodium hypochlorite solution, soaking the brown rice in a hydrophobic emulsion for airing, soaking the brown rice in a water-in-oil emulsion, taking out the brown rice, cleaning the brown rice with deionized water, culturing and germinating in a constant-temperature and constant-humidity box, drying, cooling to room temperature, and packaging in vacuum. Screened brown rice is soaked in a sodium hypochlorite solution, the brown rice is taken out, washed and then soaked in a hydrophobic emulsion to be aired, a hydrophobic porous film is formed on the surface of the brown rice, then the brown rice is soaked in a water-in-oil emulsion, an oil phase is prepared from cinnamyl aldehyde / L-glutamine cross-linked substance, carnauba wax and polyglycerol fatty acid ester, and the oil phase is prepared from a water-in-oil phase; the water phase is prepared from a staphylococcus xylosus seed solution, a Kluyveromyces marxianus seed solution and a papain solution, and the water-in-oil emulsion permeates into pores of the hydrophobic porous film and acts on the brown rice through pore channel diffusion, so that the cortex barrier of the brown rice is effectively degraded, the moisture and oxygen permeability is improved, and the germination rate and the uniformity are improved.

This invention relates to the field of microbial technology, specifically to Kluyveromyces martensii YF51-1 and its application in semi-dry fermented whey wine. This strain can efficiently ferment lactose and galactose, while also exhibiting a high ABTS scavenging rate. The fermentation broth contains a high amount of amino acid nitrogen. Under conditions where sucrose is present, the amino acid nitrogen content in the sweetened whey fermentation broth is significantly higher than that in the unsweetened whey fermentation broth. Through process optimization, the optimal fermentation conditions for strain YF51-1 were obtained, ultimately producing a semi-dry whey fermented wine with a rich aroma and a sweet and sour taste, achieving an amino acid nitrogen content of 0.337 g / L, significantly higher than existing technologies. Through optimization of the freeze-drying process, a freeze-drying protectant suitable for strain YF51-1 was derived. This protectant consists of mannose, corn steep liquor powder, lysine, and calcium chloride. After optimization, this protectant demonstrates good protective effect on the strain, with a survival rate of up to 97.22%.

The invention relates to the technical field of microorganisms, in particular to Kluyveromyces marxianus YF5-1 and application thereof in semi-dry type fermented whey wine.The Kluyveromyces marxianus YF5-1 can efficiently ferment lactose and galactose and has the high ABTS clearance rate, fermentation liquor contains much amino acid nitrogen, and under the condition that cane sugar exists, the content of the amino acid nitrogen in the fermentation liquor is increased, and the yield of the Kluyveromyces marxianus YF5-1 is increased. The content of amino acid nitrogen in the sugar-added whey fermentation liquor is obviously higher than that of sugar-free whey fermentation liquor; through process optimization, the optimal fermentation condition of the strain YF51-1 is obtained, the semi-dry whey fermented wine which is rich in fragrance, sour, sweet and delicious is finally produced, and the amino acid nitrogen of the semi-dry whey fermented wine can reach 0.337 g / L and is obviously higher than that in the prior art; a freeze-drying protective agent suitable for the strain YF51-1 is obtained by optimizing a freeze-drying powder process, the protective agent is composed of mannose, corn steep liquor powder, lysine and calcium chloride, and after optimization, the protective agent has a good protective effect on the strain, and the survival rate can reach 97.22%.

The present invention relates to a novel fermentation process using a particular strain of Kluyveromyces marxianus, referred to as KMPROTEO 1, to obtain a single-cell protein useful in the food industry.In the present description, the invention discloses a process for fermenting dextrose and other consumables to obtain a single-cell biomass. The disclosed process enables a high-protein product to be obtained that is in the range of 50 to 60% for use in the food industry. The disclosed process uses dextrose as a propagation medium for use by Kluyveromyces marxianus to obtain a single-cell biomass and, subsequently, a single-cell protein.

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 invention discloses kluyveromyces marxianus grx168 and application thereof, the kluyveromyces marxianus is preserved in the general microbiological center of the China Committee for Culture Collection of Microorganisms on June 9, 2025, and the preservation number is CGMCC NO.34816. The kluyveromyces marxianus grx168 can be used for preparing the kluyveromyces marxianus. The strain is screened from Kefir grains in Xinjiang, and a nerve cell cAMP experiment verifies that the opioid activity of a fermentation product of the strain is obviously higher than that of other yeast strains, and the cAMP content of the fermentation product of the strain is obviously lower than that of a blank control. The strain can be used for preparing fermented food or health care products with opioid activity, and a high-quality strain resource is provided for development of functional fermented products.

The invention belongs to the technical field of biological medicine, and discloses a method for preparing porcine parvovirus VP2 nanoparticles by auxotroph kluyveromyces marxianus. The dual-replicon expression vector applicable to the kluyveromyces marxianus is prepared by the applicant through screening and assembling, the vector realizes the effects of high conversion rate and high expression efficiency in the kluyveromyces marxianus, and the dual-replicon expression vector pGKD32 is shown as SEQ ID NO.2. The Kluyveromyces marxianus expression system provided by the invention is utilized by the applicant to successfully express the porcine parvovirus VP2 in a nanoparticle form, and a new thought is provided for research and development of a porcine parvovirus oral immunization strategy.

The invention belongs to the technical field of wine brewing, and discloses a low-alcohol wine brewing method applying a double-bacterium system to perform low-temperature staged fermentation, which comprises the following steps: accurately controlling the alcoholic strength to 3.8% ABV + / -0.1% ABV and ester substances to be more than or equal to 75mg / L through a heat-resistant Kluyveromyces cerevisiae and Kluyveromyces pichia composite strain system in combination with beta-glucan enzymolysis and low-temperature staged fermentation; the flavor substance retention rate is greater than or equal to 95%; wherein the Kluyveromyces thermotolerant firstly utilizes glucose to grow, breed and produce acid, so that a conversion precursor of sugar to alcohol is reduced; the kluyveromyces pichia is regulated and controlled by a metabolic pathway, most glucose is used for synthesizing flavor substances such as esters and higher alcohols, and only a small amount of glucose is converted into alcohol. The finally obtained low-alcohol grape wine has fresh and cool taste and rich fragrance, and meets the market requirements of healthy drinks. Under the joint cooperation of fermentation control, strain combination and enzymolysis treatment, the problems of out-of-control sugar conversion, flavor loss, fermentation stagnation and the like in the existing low-alcohol wine production are solved.