200 results about "Kluyveromyces marxianus" patented technology

It includes the stages of grinding the lignocellulosic biomass to a size of 15-30 mm, subjecting the product obtained to steam explosion pre-treatment at a temperature of 190-230° C. for between 1 and 10 minutes in a reactor (2), collecting the pre-treated material in a cyclone (3) and separating the liquid and solid fractions by filtration in a filter press (9), introducing the solid fraction in a fermentation deposit (10), adding a cellulase at a concentration of 15 UFP per gram of cellulose and 12.6 International Units of β-glucosidase enzyme dissolved in citrate buffer pH 4.8, inoculating the fermentation deposit (10) with a culture of the heat-tolerant bacteria Kluyveromyces marxianus CECT 10875, obtained by chemical mutagenesis from strain DER-26 of Kluyveromyces marxianus and shaking the mixture for 72 hours at 42° C.

The invention relates to a method for preparing a freeze-dried microbial agent of kluyveromyces marxianus and application of the freeze-dried microbial agent. The freeze-dried microbial agent is prepared by the aid of high-density fermentation culture and freeze-drying technologies. The kluyveromyces marxianus is preserved in the Common Microorganism Center of the China Committee for Culture Collection of Microorganisms, called as CGMCC for short, on April 27th, 2010, the preservation number of the kluyveromyces marxianus is CGMCC NO.3781, and the strain number is A3. The method and the application have the advantages that the viable cell number of high-density culture solution of the kluyveromyces marxianus can reach 3.2*10<8> cfu/mL at least and is higher than culture results of YEPD (yeast peptone dextrose) culture media by 1.1 order of magnitude at least; zymocyte solution is centrifuged, then bacterial sludge is collected and is added into cryoprotectant solution, the freeze-dried microbial agent can be prepared by the aid of the vacuum freeze-drying technologies, protectants for the freeze-dried microbial agent comprise maltose and skimmed milk powder, and the freeze-dried viable bacterium rate can reach 75% at least; the freeze-dried microbial agent is high in bacterium content, long in storage time, convenient to use and free of pollution, is environmentally friendly and can be used for fermenting milk beer and cheese and making whey wine, fruit vinegar, fruit juice and the like, processes for preparing the freeze-dried microbial agent are simple, and the like.

The invention discloses a preparation method of natural spice 2-phenethyl alcohol, which is implemented by selecting Candida drusei, Saccharomyces cerevisiae, Kluyveromyces marxianus, inoculating cultivated seed liquid or active dried yeast powder into fermentation culture medium, taking L-PHE produced by fermentation method or enzyme method as substrate, performing liquid aerobic fermentation in a fermentation shake flask or a fermentation tank, adding solid phase absorption medium, L-PHE and glucose in the process of fermentation, obtaining the solid phase absorption medium in a filtration or centrifugation manner, then adopting an elution solvent to carry out elution on the absorption medium, further distilling the eluent to obtain the natural 2-phenethyl alcohol with purity of 97.5-99.5%. The invention combines the fermentation and absorption separation of 2-phenethyl alcohol, has low production cost and excellent aroma quality of the product.

The invention provides a recombinant vector used for carrying out foreign gene secretory expression in an auxotrophic kluyveromyces marxianus strain as well as a preparation method and application of the recombinant vector. The recombinant vector comprises ampicillin resistance genes, a PKD1 vector, inulase promoters, signal peptides, multiple cloning sites, inulase terminators, nutritional gene promoters and a nutritional gene open reading flame according to the sequence. The constructed recombinant vector used for carrying out foreign gene secretory expression and the preparation method of the recombinant vector can be used for constructing a transformant to achieve secretory expression of foreign genes.

The invention relates to a method for preparing cholesterol reducing eggnog fermenting drink by utilizing lactobacillus and microzyme which produce bile salt hydrolase, belonging to a functional drinkwhich is prepared from the following ingredients: 15% of whole egg pulp, 35% of milk, 7% of saccharose, 0.3% of CMC-Na. and 0.2% of carrageenin, wherein the ratio of the raw materials (the whole eggplug and the milk) to water is 1:1 (W/V). The primary fermentation condition is described as following: streptococcus thermophilus Tx, Lactococcus lactis subsp.lactis KS4 and Lactobacillus casei KL1 are mixed in proportion of 1:1:1, the inoculum size is 3%, and then the mixture is fermented for 8h at the temperature of 43 DEG C. The secondary fermentation condition is described as following: Kluyveromyces marxianus M3 and K1 are compounded in proportion of 1:1 or the inoculum size of the single strain is 1%, and then the composite is fermented for 24-26 h at the temperature of 30 DEG C (mixture strain) or 32-34 DEG C; after ripening is carried out for 1 day at the temperature of 4 DEG C, the reducing rate of raw material cholesterol reaches as high as 89.3%. By utilizing the characteristics of strain high-yielding BSH, exopolysaccharide and the like, the invention has the efficiencies of immunological regulation, cholesterol reducing, tumor prevention and the like, and the problem of egg smell is not only solved, but also the content of raw material cholesterol is efficiently reduced through egg pulp proper sterilization and eggnog secondary fermentation. The eggnog drink preparedby the invention has simple fermentation process, short period and lower cost and is suitable for industrialized production.

The invention provides a kluyveromyces marxianus engineering bacterium obtained from recombinant expression of porcine circovirus type II capsid protein; and porcine circovirus type II virus-like particles are obtained by cloning porcine circovirus type II capsid protein genes into an expression vector and conducting recombinant expression in kluyveromyces marxianus. The invention also provides a preparation method of a porcine circovirus type II virus-like particle vaccine, wherein the preparation method comprises the following steps: preparing the porcine circovirus type II virus-like particles through high-density fermentation of the recombinant kluyveromyces marxianus, and then conducting separation and purification so as to prepare the vaccine. The porcine circovirus type II virus-like particle vaccine provided by the invention, which can generate a high-level serum IgG antibody just by implementing injection immunization once, has the advantages of being good in immunizing effect, high in safety, simple in culture operation, high in yield, low in production cost and the like, and the porcine circovirus type II virus-like particle vaccine can achieve large-scale enlarged production; and the porcine circovirus type II virus-like particle vaccine can be used for relieving and preventing related clinical symptoms caused by the infection of porcine circovirus type II (PCV2).

The invention discloses a composite microbial preparation and a preparation method and application thereof. The microbial preparation contains Enterococcus faecium SC13 with an accession number of CCTCC NO: M2014060 and Kluyveromyces marxianus C2 with an accession number of CCTCC NO: M2014059 which are separated by the inventor. Enterococcus faecium SC13 and Kluyveromyces marxianus C2 are combined with Bacillus amyloliquefaciens, jerusalem artichoke powder and food-grade diatomite; through cooperation among bacteria and between the bacteria and a filler, the immune system of an animal is strengthened, proliferation of beneficial bacteria in the intestines of the animal is selectively promoted, colonization and propagation of harmful bacteria are inhibited, the microbial environment in the body of the animal is improved, immune response of organisms is stimulated, immunity and production performance of livestock and poultry are enhanced, usage of antibiotics is reduced, good recognition adhesion and elimination capability on mycotoxins in a feed is obtained, and the conversion rate of the feed is increased.

The invention discloses a Kluyveromyces marxianus C2 and a preparation method and an application thereof. The preservation number of the Kluyveromyces marxianus C2 is CCTCC NO: M2014059. The aflatoxin B1, zearalenone and ochratoxin A can be degraded by Kluyveromyces marxianus C2 and the biodegradation rate reaches more than 80%. The fermented Kluyveromyces marxianus C2 is put into toxin-contaminated liquid or solid food or feed matrix at a certain ratio to enable mycotoxins to be fast, safely and efficiently degraded and detoxified and can also be used for oral detoxification of animals. The strain disclosed by the invention has the advantages of low production cost and safe application and has wide application prospects.

The invention relates to a sunflower seed meal microbial protein feed. The sunflower seed meal microbial protein feed is prepared by the following steps: mixing sunflower seed meal and wheat bran in a weight part ratio of 80 to 20 and adding water to prepare a solid-state fermentation culture medium; inoculating 5 parts by weight of a mixed strain on the solid-state fermentation culture medium and mixing and fermenting at 20-32 DEG C; and after fermenting, drying and crushing a fermented product to obtain the feed. The sunflower seed meal microbial protein feed takes the sunflower seed meal with husks as a main raw material, so that the production cost is low. The sunflower seed meal microbial protein feed contains bacillus subtilis, kluyveromyces marxianus, lactobacillus plantarum and candida tropicalis, has no any chemical additive and has the mellow flavor of fermented natural plants; the crude protein content of the feed is 35.5% and a plurality of amino acids, vitamins and microelements are contained in the feed, so that the sunflower seed meal microbial protein feed is a high-quality and low-price nutritional microbial protein feed.

The invention relates to a eukaryon expression method for exoinulinase, which mainly comprises the following steps: firstly, connecting a nucleotide sequence (as shown in SEQ ID No: 1) of the exoinulinase of Kluyveromyces marxianus with pichia expression plasmids pPICZ alpha A through a restricted enzyme site, and obtaining a recombinant expression vector; and secondly, introducing the recombinant expression vector into a thermococcus host strain X-33 or SMD1168, and expressing target proteins (the Kluyveromyces marxianus exoinulinase) through induction fermentation. The eukaryon expression method for the exoinulinase lays a foundation for developing the exoinulinase with industrial application value.

The present invention provides an application of kluyveromyces marxianus yeast, especially the applications in feed additives and feed fermentation, and relates to a feed and a feed additive. The kluyveromyces marxianus yeast is preferably used as an animal feed additive and/or an animal feed fermentation agent, and can improve the immune system of animals, and/or improve the digestion ability of animals, and/or protect animals from free radical damage, and/or produce more nutrients. More preferably, compared to conventional yeasts, the kluyveromyces marxianus yeast can better improve animal immunity, and/or improve animal digestion, and/or protect animals from free radical damage, and/or produce more nutrients.

The invention provides a preparation method of whey nutritional wine. The whey, a byproduct of cheese, is used as the raw material; the whey is sterilized under high temperature and cooled before being inoculated with 8-10% of bacillus subtilis zymotic fluid; the mixture is hydrolyzed for 30-36 hours at the preserved temperature of 30 DEG C, the mixture is sterilized and the temperature thereof is lowered to 25 DEG C; glucose is added to the mixture to adjust total sugar content of the zymotic fluid to 11-13%, the mixture is inoculated with 3-5% Marx Clewett microzyme suspension, heat preservation anaerobic fermentation is carried out on the mixture for 18-20 hours; then high-speed centrifugation deslagging and membrane separation are carried out on the mixture before supernatant fluid is extracted; then low temperature vacuum concentration is carried out on the mixture for alcohol reflows; finally, the mixture is blended before preparing milone of various alcohol contents.

The invention provides a recombinant vector for expressing a histidine-tag-fused foreign gene in a Kluyveromyces marxianus nutritional deficient strain, and a preparation method and application of the recombinant vector. The recombinant vector sequentially comprises an ampicillin resistance gene, a PKD1 vector sequence, an inulase promotor, multiple cloning sites, an inulase terminator, a nutritional gene promotor and a nutritional gene open reading frame, and further comprises histidine tags located at C ends or N ends of the multiple cloning sites. The recombinant vector provided by the invention can be used for building a transformant to realize expression of the foreign gene.

The invention belongs to the technical field of biological medicine, and concretely relates to a swine fever virus subunit vaccine and a preparation method and a purpose thereof. The invention provides a kluyveromyces marxianus yeast recombination strain used for preparing the swine fever virus subunit vaccine. The recombination strain is constructed by the following steps: swine fever virus envelope protein E2 is intercepted, through codon optimization, a coding sequence of the swine fever virus mE2 protein is obtained, and then is cloned to a kluyveromyces marxianus yeast expression vector,and the kluyveromyces marxianus yeast host strain is transformed. The invention also provides the method for preparing the swine fever virus subunit vaccine, which comprises the following steps: the kluyveromyces marxianus yeast host strain is subjected to recombination expression by using mE2, steps of culture, centrifugation, cell disruption, and separating purifying are carried out to obtain the swine fever virus mE2 protein antigen, and the purified antigen and an adjuvant are subjected to complex formulation to prepare the swine fever virus subunit vaccine. The injection immunotherapy ofswine fever virus mE2 protein recombination subunit vaccine can obtain a protective IgG antibody, and the subunit vaccine can reduce and prevent the swine fever virus infection-related disease.

The invention provides a feruloyl esterase gene. The feruloyl esterase gene is originated from microbes in the rumen of a Chinese yak and has a nucleotide sequence as shown in SEQ ID No. 1, wherein the nucleotide sequence codes an amino acid sequence as shown in SEQ ID No. 2. The invention also provides a recombinant Kluyveromyces marxianus expression vector containing the gene, a genetically engineered strain, a preparation method for feruloyl esterase and application of prepared feruloyl esterase. Feruloyl esterase is subjected recombinant expression in a Kluyveromyces marxianus expression system; and after high-density fermentation for 48 h, intracellular and extracellular feruloyl esterase vitalities are 686.35 U/mL and 346.34 U/mL, respectively. Feruloyl esterase having undergone recombinant expression by Kluyveromyces marxianus can release ferulic acid in corn bran. Feruloyl esterase prepared in the invention can be applied to food processing, feed addition, biomedicine, biotransformation of cellulosic substances, bioenergy and other fields.

The invention belongs to the field of environmental protection and particularly discloses aa microbial deodorant for deodorization. The microbial deodorant comprises the active components: a microbial agent, molasses, amino acids and vitamins; the microbial agent is a composite bacterial agent of Kluyveromyces marxianus and Bacillus subtilis; the microbial deodorant is capable of inhibiting the growth of putrefying bacteria, improving the degrading means of organic matters and decreasing the release of ammonia and hydrogen sulfide and occurrence of ammonia matters, and is capable of deceasing methane from bacterial decomposition, thereby arriving at deodorization; the materials of the microbial deodorant are easy to obtain, the preparation method is simple, and the microbial deodorant is worthy of popularization and application.

The invention relates to the technical field of fermented dairy products, and discloses multi-bacteria fermented skim camel milk for resisting diabetes and a production method of the multi-bacteria fermented skim camel milk. The multi-bacteria fermented skim camel milk is prepared from a lactobacillus helveticus leavening agent, a lactobacillus plantarum leavening agent, a lactobacillus paracasei leavening agent, a lactobacillus paracasei hard subspecies leavening agent, a lactobacillus mucosae leavening agent, a Harbin lactobacillus leavening agent, a lactobacillus hilgardii leavening agent, a lactobacillus pentosus leavening agent, a lactobacillus rhamnosus leavening agent, a lactococcus lactis leavening agent, an issatchenkia orientalis leavening agent, a kluyveromyces marxianus leavening agent, a pichia membranefaciens leavening agent and an ethanol candida leavening agent, which are added to the skim camel milk and are fermented. The multi-bacteria fermented skim camel milk is good in taste, the production method is easy to operate and simple in operation method; the obtained multi-bacteria fermented skim camel milk is abundant in nutrient, contains the components such as probiotics, immunomdodulating peptide, anti-oxidative peptide and the like, and has antidiabetic, anti-inflammation and antioxidant effects; and the physique and the overall vitality of people can be enhanced.

The invention discloses construction and application of xylitol high-temperature and high-yield engineered strains. Xylose reductase genes from different sources are efficiently expressed in heat-resistant yeast Kluyveromyces marxianus through a gene engineering method, so that the strain can be used for producing a large amount of xylitol by efficiently using and fermenting xylose under higher temperature (which is higher than 42 DEG C) through a xylose reductase metabolism path. Heat-resistant engineered yeast strains YZJ015 and YZJ017 capable of using xylose to carry out fermentation are constructed, and the preservation numbers are respectively CGMCC No.7819 and 7820. The yeast strains can be applied to produce xylitol with higher yield and higher production rate under the temperature of 42-45 DEG C by fermentation through different xylose concentrations. Furthermore, the strain YZJ017 (CGMCC No.7820) can also be used for preparing xylitol from glycerinum and xylose through fermentation.

The invention discloses a bacterial strain producing beta-galactosidase and a method for preparing high-purity galactooligosaccharide. The bacterial strain producing beta-galactosidase is pichia pastoris SMD1168H-pGAZaA-lac, and the preservation number is CGMCC No. 8349. The invention also discloses the method for preparing high-purity galactooligosaccharide. The method includes the following steps: using pichia pastoris strain SMD1168H-pGAZaA-lac to prepare beta-galactosidase; using beta-galactosidase to catalyze lactose, so as to synthesize galactooligosaccharide; purifying galactooligosaccharide mixed liquid through Kluyveromyces marxianus; refining to obtain galactooligosaccharide with high purity. The purity of galactooligosaccharide obtained through the method is equal to or greater than 90%, and the yield is equal to or greater than 30%.

The invention discloses Lactobacillus plantarum WZMX-2 as well as application and prepared whey pear wine thereof. Bitter-free whey pear wine is prepared by the following steps: uniformly mixing whey fermentation liquor composed of Lactobacillus plantarum WZMX-2, Kluyveromyces marxianus WWMJ-1 and saccharomyces cerevisiae at the volume ratio of 4:4:2 with whey bergamot pear juice mixed liquor; culturing at 25-37 DEG C for 24-72 hours; and fermenting when measuring that the alcohol degree reaches 7.8% vol, the total acid reaches 60.84 titration acidity, the total sugar content reaches 42.34 g/L, and the bitter peptide content reaches 0.15 g/100 mL. The prepared whey pear wine is yellowish green, clarified and translucent, has natural color and fragrance of whey and pear, and has no bitter feeling; functional tests prove that the prepared whey pear wine can prolong weight-bearing swimming time of mice, reduces the generation of serum urea, increases the reserve of liver glycogen, has no apparent influence on weight and blood lactic acid of the mice, and has a functional effect of alleviating physical fatigue.

The invention discloses an acetic acid and xylose co-used Kluyveromyces marxianus strain with a biological preservation number of CGMCC No.16757. The strain is obtained by long-term orient domestication of Kluyveromyces marxianus through cellulosic hydrolysate and is prepared by the following steps: picking single yeast colonies from a YPD flat plate and carrying out 40 DEG C overnight culture; reinoculating collected cells to a fresh culture medium containing the cellulosic hydrolysate with certain concentration, enabling an optical density value of the culture medium at 620nm to be approximate to 0.3; culturing for 24 to 48 hours at 40 DEG C and 150rmp/min; when OD620 of the acetic acid and xylose co-used Kluyveromyces marxianus strain is greater than or equal to 2.0 (5 to 7 generations), collecting the cells; repeating the steps until the growth speed of the cells is obviously improved under the stress condition of an inhibitor with the concentration; improving the concentration ofthe cellulosic hydrolysate and starting new repeated culture; after continuous domestication, coating the YPD flat plate containing the cellulosic hydrolysate with the collected cells and culturing at40 DEG C for 24 to 48 hours; picking the single colonies with good growth situation and putting into a YPD liquid medium, and culturing at 40 DEG C and 150rmp/min for 24 to 48 hours; preserving dominant resistance clones in 30 percent glycerinum.

The invention relates to a walnut milk fermentation agent which is applied to fermentation for preparing a walnut milk lactobacillus fermentation beverage. The walnut milk fermentation agent comprises lactobacillus plantarum subspecies, Pediococcus pentosaceus and kluyveromyces marxianus. Meanwhile, the invention also relates to a method for preparing the walnut milk lactobacillus fermentation beverage by utilizing the walnut milk fermentation agent and the walnut milk lactobacillus fermentation beverage prepared by utilizing the method. The walnut milk lactobacillus fermentation beverage which is favorable in taste, mouthfeel and quality and has dual functions of walnuts and fermentation food is prepared by carrying out inoculated fermentation on the walnut milk by the walnut milk fermentation agent. The fermentation process is simple without strain domesticating, and the stability of quality of the fermented milk can be ensured.

The invention provides a kluyveromyces marxianus with a preservation number of CGMCC No. 10621, and also provides the application of the kluyveromyces marxianus. Compared with existing kluyveromyces marxious such as ATCC8585 and ATCC26548, the kluyveromyces marxianus has higher protein secretion capacity and higher biomass, can be used for producing enzymic preparations, and also can be used for building a recombinant protein expression system.

In view of problems in the existing cellulose ethanol preparation technologies that inhibitors in a cellulose hydrolyzate inhibit fermentation, xylose cannot be completely converted, glucose inhibitsfermentation of the xylose, and a cellulase acting temperature and a microbe fermentation temperature are non-uniform, the invention provides a fermentation method for producing xylitol and ethanol byusing undetoxified cellulose raw materials. The fermentation method comprises the steps of inoculum culture, xylose fermentation, cellulase adding, synchronous diastatic fermentation and the like. According to the method, simultaneous utilization of the xylose and the inhibitors is achieved by adopting an acetic acid and xylose co-utilization based Kluyveromyces marxianus strain, and thus, the problem in inhibitor removal is avoided; by adopting high-temperature synchronous diastatic fermentation, the problem that the cellulase is unmatched with a fermentation temperature is avoided; and compared with the existing cellulose ethanol fermentation technologies, the production cost can be remarkably reduced.