77 results about "Genus Kluyveromyces" patented technology

The invention discloses an alcohol dehydrogenase, a gene thereof, a recombinant expression vector and a recombinant expression transformant respectively containing the gene, a recombinase of the alcohol dehydrogenase, and an application of the alcohol dehydrogenase in asymmetric reduction synthesis of chiral diaryl secondary alcohol as a catalyst, and belongs to the technical field of bioengineering. The alcohol dehydrogenase is from Kluyveromyces sp. CCTCCM2011385, has a carbonyl group reduction function, and also has a hydroxy group oxidation function. Extra addition of glucose dehydrogenase and other enzymes used for cofactor circulation is not needed when the alcohol dehydrogenase is used in the reduction of diaryl ketone into the chiral diaryl secondary alcohol as a biocatalyst, and the alcohol dehydrogenase has the advantages of high catalysis efficiency, mild reaction conditions, easy product recovery and low cost, so the alcohol dehydrogenase has very good application and exploitation prospect in the production of antihistamine medicines.

The invention discloses a composite strain starter and the preparation method, belonging to the microorganism fermentation field, which is mainly prepared with equivalent lactobacillus delbrueckii Bulgaria subspecies, lactococcus lactis, Leuconostoc mesenteroide, lactobacillus acidophilus, lactobacillus casei, kluyveromyces and brettanomyces. The preparation method is as follow: the strains are optimized and cultivated, and then are respectively fermented, centrifuge-treated, concentrated, added with cell protective agent, and freezed in vacuum. The invention is compounded with various functional strains; the acidophilus milk made by the omposite strain starter has the advantages of original flavor, which has the function indexes close to kefir particle fermentation acidophilus milk, and can be excellent direct distribution kefir starter; the fermented acidophilus milk has strong inhibition effect to pathogenic microorganism, and has good therapeutic effect to gastrointestinal diseases, metabolic disease, hypertension and heart disease.

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 belongs to the biotechnology field, in particular to a method of manufacturing ethanol by utilizing Jerusalem artichoke material. The method comprises the following steps: in material process step, the fresh Jerusalem artichoke is cleaned and juiced, or sliced up, dried and pulverized; in a seed liquid preparation step, kluyveromyces are cultivated in seed culture fluid taking Jerusalem artichoke powders or Jerusalem artichoke juice as a complete composition; in a ferment step, seed liquid is inoculated in culture medium confected by the complete Jerusalem artichoke powders or the complete Jerusalem artichoke juice, the ethanol is produced through anaerobic fermentation, and the ethanol is obtained by distillation at last. Both compositions of a seed culture medium and a ferment culture medium provided by the method are Jerusalem artichoke powders or Jerusalem artichoke juice; any auxiliary composition is not needed; and beforehand saccharification processing of Jerusalem artichoke material in the ferment culture medium is not needed; inulase excreted by the kluyveromyces in the seed liquid directly hydrolyzes and saccharifys multi fructosan in the Jerusalem artichoke material; the ferment is processed at the same time; and concentration of ethanol product is high to 12 percent (v/v). The method has the advantages that the method is simple; the synchronization process of saccharification and ferment is implemented; and the production cost is reduced.

A process is provided for the bioconversion of a carbon substrate to 1,3-propanediol by a single organism utilizing microorganisms, such as, Citrobacter, Enterobacter, Clostridium, Klebsiella, Aerobacter, Lactobacillus, Aspergillus, Saccharomyces, Zygosaccharomyces, Pichia, Kluyveromyces, Candida, Hansenula, Debaryomyces, Mucor, Torulopsis, Methylobacter, Escherichia, Salmonella, Bacillus, Streptomyces and Pseudomonas, containing the genes encoding for an active glycerol or diol dehydratase enzyme by contacting these organisms with a carbon substrate under the appropriate fermentation conditions. Specifically, Citrobacter and, Klebsiella provide the source of exogenous genes for such active dehydratase enzyme.

The invention relates to a micro-ecological composite fungus preparation and a method for preparing the same, in particular to a micro-ecological preparation and a method for preparing the same. The method solves the problems that the prior micro-ecological preparation can only temporarily increase and supplement the amount of probiotics in intestinal canal of a human body in a short period, and has poor competitiveness and viability so that the probiotics are used up after a period of time of stopping taking the micro-ecological preparation. The micro-ecological composite fungus preparation mainly comprises bacillus natto, lactic acid kluyveromyces, lactobacillus acidophilus and soybean oligosaccharides. The method comprises the following steps: culturing micro-ecological funguses; intermediately culturing the micro-ecological funguses; culturing and centrifuging the micro-ecological funguses after the micro-ecological funguses are transferred into a high concentration culture fluid; and adding auxiliary materials to the micro-ecological funguses, pre-freezing and vacuum freeze-drying the mixture, and adding the soybean oligosaccharides to the mixture to obtain the micro-ecological composite fungus preparation. The total number of viable beneficial bacteria in the micro-ecological composite fungus preparation is between 1.0 x 10<10> and 9.9 x 10<10> per gram, the micro-ecological composite fungus preparation not only has huge unit number, but also has high survival rate of the viable bacteria of more than 98 percent two years later.

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 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).

A process is provided for the production of an antibody or a fragment or functionalized fragment thereof using a transformed lower eukaryotic host containing an example DNA sequence encoding the antibody or (functionalized) fragment thereof, wherein the antibody or (functionalized) fragment thereof is derived from a heavy chain immunoglobulin of Camelidae and is devoid of light chains, and wherein the lower eukaryotic host is a mould, preferably belonging to the genera Aspergillus or Trichoderma, or a yeast, preferably belonging to the yeast genera Saccharomyces, Kluyveromyces, Hansenula, or Pichia. The heavy chain fragment can contain at least the whole variable domain. A complementary determining region (CDR) different from the CDR belonging to the natural antibody ex Camelidae can be grafted on the framework of the variable domain of the heavy chain immunoglobulin. The catalytic antibodies can be raised in Camelidae against transition state molecules. The functionalized antibody or fragment thereof can comprise a fusion protein of both a heavy chain immunoglobulin from Camelidae or a fragment thereof and another polypeptide, e.g., an enzyme, preferably an oxido-reductase. Also provided are new products obtainable by a process as described, and compositions containing a product produced by a process as described, which composition may contain a new product as provided.

The invention belongs to the technical field of gene engineering, in particular to a technical proposal used to improve the expression level of recombinant protein. In the method, a promoter, alpha-signal peptide, target protein of an inulase gene are orderly connected with a terminal subcode sequence of the inulase gene first; then the sequence is inserted into a Kluyveromyces lactis expression vector; finally the Kluyveromyces lactis is transformed and fermented, and a fermented supernatant is taken and separated. The method can accelerate the remarkable growth of the yield of the recombinant protein expressed in the Kluyveromyces lactis. The technical proposal can be used to improve the expression amount of various proteins and provide a new method for reducing production cost and enlarging production scale.

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 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 present invention relates to a composite bacteria milk product and its preparation method. It is characterized by that the composite bacterial culture composed of a fungus (lactic acid kluyveromyces) and a life-benefiting bacterium (lactobacillus delbruckii bulgaricus subspecies) can be inoculated in the sterilized milk so as to obtain the invented milk product with uniform gel, palatable taste and several health-care functions of regulating stomach and intestine. Said invention can do not use any food additives.

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 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 a recombinant yeast Kluyveromyces for expressing antibodies or antibody analogues and application thereof. The invention provides a method for constructing the recombinant yeast Kluyveromyces for expressign antibodies or antibody analogues. The method comprises the following steps: encoding genes of the antibodies or the antibody analogues are guided into the yeast Kluyveromyces to obtain recombinant bacteria; the antibody analogues are fusion proteins formed by Fc fragments of the antibodies and proteins or protein subunits; the encoding genes of the antibodies are heavy chain and light chain encoding genes of the antibodies. The recombinant yeast Kluyveromyces for expressing antibodies or antibody analogues can be cultured to produce complete antibodies or antibody analogues. The invention uses the yeast Kluyveromyces to prepare the antibodies or antibody analogues to preliminarily overcome the difficulties of difficult antibody production, high purity and large quantity requirements for products and provide good application prospects.

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 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.