206 results about "Saccharomycotina" patented technology

The invention discloses straw and pot ale mixed fermented feed and a production method thereof, and relates to the field of environmental science, in particular to the mixed fermented feed and the production method thereof. The production method comprises the following steps: (1) crushing straw, and then mixing pot ale; (2) preparing aerobic fermentation bacteria liquid, uniformly spraying the aerobic fermentation bacteria liquid into materials, adding compound trace elements, and fermenting; (3) preparing anaerobic fermentation bacteria liquid, appropriately adding according to the fermentation situation of the fermented materials, and fermenting; and (4) drying the fermented materials containing water, and then getting the straw and pot ale mixed feed. Aerobic bacteria used in the method are high-efficient cellulose and lignin degrading bacteria, thereby improving the degradation rate of cellulose and lignin, fully utilizing yeast and other microbes in the pot ale to produce single-cell proteins, improving the content of nutrition in the feed, enabling the protein content to be 13-20%, having certain bouquet and acid fragrance and improving the palatability of the feed. The fermentation process is simple to operate, the fermentation time is 7-9 days, and the production time of the feed is shortened.

The invention discloses a CRISPR/Cas9-mediated large DNA fragment assembling method. Specifically, the invention provides a nucleic acid construct capable of carrying out constitutive expression of Cas9 in yeast; the nucleic acid construct contains a yeast Tef1 promoter operationally connected with a Cas9 gene, a replication origin from pBR322 and a screening mark thereof, and a replication region from CEN6ARS4 and a screening mark thereof; and the nucleic acid construct is in the form of single-copy replication in the yeast and in the form of high-copy replication in Escherichia coli. The invention also provides a nucleic acid construct capable of carrying out constitutive expression of sgRNA in yeast, a nucleic acid construct used as a receptor vector and a DNA assembling method. According to the invention, two or more to-be-assembled large DNA fragments are successfully assembled in microzyme to form a large plasmid in one shot; so low-efficiency in-vitro digestion recovery, genetic transformation and the like of large DNA fragments are avoided, and the method provided by the invention is more convenient and efficient compared with traditional methods.

The utility model relates to a method for producing a live yeast cell derivative, which comprises following steps: selecting yeast as a bacteria source, adopting wort as inclined medium, adopting YEPD medium which is suitable for the yeast to grow as liquid medium to culture liquid spawn, culturing the liquid spawn to a logarithmic phase 8 hours-12 hours, obtaining yeast culture medium, transforming the yeast culture medium from the most suitable temperature to the high temperature of 37 DEG C, stressing for 30 minutes, adopting a refrigerated centrifuge to separate the yeast culture medium centrifugally, collecting yeast thalli, adopting a high pressure homogenizer to crush yeast thalli cell wall, centrifugally separating the yeast cells for the second time, removing sediment, and obtaining the live yeast cell derivative. The method does not adopt a traditional protease hydrolysis method to crush the wall but selects the high pressure homogenizer to crush the yeast cells, the crushing effect is good, the operation is convenient, the method is easily extended and applied and can realize industrialization, and simultaneously the method has the advantages of low production cost, little investment and strong practicability.

The invention discloses a method for processing sludge to prepare biological organic fertilizer and a special starter thereof. The active components of the sludge starter are composed of bacillus subtilis, bacillus stearothermophilus, aspergillus niger, thermophilic sporotrichum and saccharomyces cerevisiae. The provided method for processing sludge to prepare biological organic fertilizer is as follows: the sludge and excipient are mixed; then the provided sludge starter is inoculated into the mixture of the sludge and the excipient to ferment the mixture of the sludge and the excipient, and thus matured sludge is obtained; finally, streptomyces microflavus is inoculated into the matured sludge to cultivate the matured sludge continuously until the biological organic fertilizer is obtained. The method for processing sludge to prepare biological organic fertilizer has the advantages of simple technical procedures, less investment on the equipment, and good fermentation, dehydration, deodorization and maturity effects. Thus, the method for processing sludge to prepare biological organic fertilizer is a practical technique that makes the sludge harmless, makes the sludge become resource and recycles the sludge.

The invention discloses a micro-ecological flora of crop root system probiotics and an application thereof. The micro-ecological flora comprises a leading flora, a balance flora, a basic flora, a secondary-function flora and a special-function flora, wherein the leading flora comprises 25-35 parts of nitrogen-fixing bacteria by mass; the balance flora comprises 17-23 parts of phosphate solubilizing bacteria and 17-23 parts of potassium solubilizing bacteria by mass; the basic flora comprises 2-7 parts of yeast, 2-7 parts of rhizobia and 2-7 parts of photosynthetic bacteria by mass; the secondary-function flora comprises 2-7 parts of bacillus and 2-7 parts of actinomycetes by mass; the special-function flora comprises 2-4 parts of white rot fungi and 1-3 parts of pseudomonas. In the micro-ecological flora disclosed by the invention, the strain stability is good, functional microorganisms can be effectively planted in soil, and the functions of improving the crop yield and quality, improving the soil, detoxifying heavy metals and resisting diseases and pests can be realized.

The invention discloses a rapid assembling method of multi-fragment DNA yeast. The rapid assembling method comprises the following steps: (1) performing co-transformation on a plurality of DNA molecules with homologous arms, and linearized yeast shuttle vectors, so as to obtain yeast; (2) eluting all the transformed yeast colonies on a whole screening culture plate, centrifuging, and discarding the supernate to obtain transformed yeast cells; (3) extracting plasmid DNA of the yeast cells obtained in step (2), transforming the competent cells of colon bacillus; and (4) screening colon bacillus, and cloning to obtain large-fragment DNA. The method for assembling large-fragment DNA molecules is fast in speed, simple, convenient and feasible, high in success rate, low in cost, high in efficiency, easy to operate, beneficial in enlarging the industrialization scale and wide in application, and a plurality of small-fragment DNA molecules can be assembled into one large-fragment DNA molecule.

The invention relates to candida chromogenic medium, a detection kit and a detection method, belonging to the technical field of microbial diagnosis, in particular to the cultivation and identification of yeast in medical microbiology specimens, drugs and medical equipment sanitary inspection samples, public health surveillance samples and food (including cosmetics) sanitary inspection samples. The chromogenic medium of the invention is composed of basic medium, mixed chromogenic substrate and bacteriostat, wherein the mixed chromogenic substrate consists of aminocaproic glucosidase and alkaline phosphatase substrate, and candida specific enzyme is added into the mixed chromogenic substrate. The detection kit of the invention consists of the chromogenic medium, identification paper A containing enzyme substrate 5-bromo-4-chloro-3-indolyl-N-acetyl-beta-D-aminogalactose and identification paper B containing enzyme substrate 5-bromo-4-chloro-3-indolyl-beta-D- glycopyranoside. In the invention, the candida chromogenic medium and the detection kit have the advantages of low cost and simple configuration, and the method can be applied to the separation and identification of candida rapidly, simply and accurately.

A beta-glucosidese, its encoding gene and use are disclosed. It consists of (a) or (b) protein, (a) protein is made of amino-acid sequence in sequence 3; (b) protein is derived by (a) and substituted and/or lost and/or added by one or several amino acid in sequence 3 and has beta-glucosidese. The process is carried out by: taking cellulose as material and synchronized mashing and common fermenting. It has better enzyme activity and can be used to produce alcohol.

The invention relates to a complex lively bacterial preparation which is prepared through the following method: (1) mixing the seed solutions of bacillus subtilis with an OD value of 3.5 to 4.0, saccharomycetes with an OD value of 3.0 to 4.0, bacillus mucilaginosus with an OD value of 3.5 to 4.0 and bacillus licheniformis with an OD value of 3.5 to 4.0 according to the volume ratio of (1 to 5):(1 to 5):(1 to 5):(1 to 5), so as to obtain a mixed seed solution;(2) carrying out high density mixed fermentation cultivation on the mixed seed solution in a by-product produced during yeast fermentation, and not stopping until the total of viable counts is larger than 1 billion/mL, so as to obtain the complex lively bacterial preparation. The complex lively bacterial preparation provided by the invention can be mixed with nutrient elements such as nitrogen and phosphorus to prepare a microorganism water soluble fertilizer, and the water soluble fertilizer prepared from the complex lively bacterial preparation is comprehensive in nutrition and can be easily absorbed by plants, the preparation raw materials of the fertilizer are cheap and easy to obtain, and the microorganism therein can inhibit the growth of pathogenic bacteria, promote the growth of crops and induce the plants to generate resistance, and takes the effects of prophylaxis and growth promoting on the crops.

The invention discloses a microbial fermentation bed strain and a manufacturing method thereof. The microbial fermentation bed strain comprises the following components in percentage by weight: 50 to 70 percent of bacillus subtilis strain and 30 to 50 percent of saccharomyces cerevisiae. The strain adopted by the invention is safe and reliable; the production is low in cost, energy-saving and environmentally-friendly and adopts simple and controllable process; the pathogenic bacteria, parasite ova, grass seed and the like in packing can be killed; the strain can decompose livestock and poultry manure quickly, optimize breeding environment and convert the livestock and poultry manure into probiotic bacterium mycoproteins used for feeding the livestock and poultry, so the material to meat (egg) ratio is lowered effectively and breeding economic benefit is improved; and in a packing fermentation process, these kinds of bacteria have a mutually beneficial and symbiotic effect on the dynamic change of the pH value and the oxygen content, so a high constant stability is ensured in the long-term operation of the fermentation bed, and the requirement on the packing is low.

The invention discloses a biological control method for enhancing the postharvest storability of strawberries. The biological control method comprises the following steps of: respectively preparing 105-108 CFU/mL hanseniaspora uvarum suspension and 105-108 CFU/mL pichia guilliermondii suspension; compounding 105-108 CFU/mL compound antagonistic yeast suspension according to isoconcentration and iso-volume; respectively carrying out spraying treatment on strawberry fruits 1-7 days before the strawberry fruits are picked by using the compound antagonistic yeast suspension; and storing after picking under the conditions of 2 DEG C and 90%-95% of RH (Relative Humidity), or selling at normal temperature. The biological control method disclosed by the invention achieves no toxic or side effect on human bodies, prolongs the storage period and shelf life of the strawberries, carries out the field spraying treatment on the strawberries by utilizing compound antagonistic yeast and can keep the better quality of the strawberries in the storage period, outstandingly enhance the commodity rate of the strawberries and the activity of a relative disease-resistant defense enzyme and reduce the rotting rate.

The invention relates to a novel gene sequence of a recombinant chicken Alpha interferon, constructs a recombinant expression vector thereof and belongs to a gene engineering biological product obtained by a molecular biology method. The gene sequence of a newly designed chicken Alpha interferon is recombined into a pPICZ Alpha-A vector and then is confirmed on the special position of a microzyme by an electric conversion mode. Besides, a pichia expression system is adopted to express a foreign gene, thus being beneficial to the commercial production of the chicken interferon. The protein expressed by a gene group after being diluted by 4365158.3 times can completely restrain the attraction of vesicular stomatitis virus of 100-1000TCID50. Compared with a natural chicken Alpha interferon, the novel gene sequence of a recombinant chicken Alpha interferon has a higher anti-virus effect; the anti-virus effect thereof is improved by 8 times. Test also detects that the protein expressed by the gene can restrain the proliferation of a newcastle disease virus and an avian influenza virus; besides, the effect of the interferon with high concentration is more remarkable.

The invention discloses recombinant yeast and a preparation method thereof. The method comprises the following steps of: (1) transferring an expression vector I into host yeast to obtain recombinant yeast transferred into the expression vector I, and recording the recombinant yeast as yeast A; and (2) transferring an expression vector II into the yeast A to obtain recombinant yeast transferred into the expression vector I and the expression vector II, and recording the recombinant yeast of the expression vector I and the expression vector II as yeast B. Experiments prove that the recombinant yeast prepared by the preparation method does not carry any resistance genes; the transferred expression vectors have multiple copy numbers between 5 and 50, and the copies are all integrated into a genome of the host yeast; and exogenous gene products expressed by the recombinant yeast are safe food-grade products. The preparation method has the advantages of simple operation, time saving, labor saving and low cost, and thus the recombinant yeast and the preparation method thereof are suitable for popularization and application.

The invention belongs to the detection technical fields of feed science and feed additives, and more specifically relates to a rapid qualification and quantitation measurement method of saccharomyces cerevisiae in an additive premix sample. According the invention, cDNA gradient dilution is taken as an external standard substance used for distinguishing dead bacteria and live bacteria for accurately detecting the amount of the live bacteria in a sample to be measured. The invention is capable of measuring saccharomyces cerevisiae in premix sample with simple, rapid and accurate qualification and quantitation without pure culture of saacharomyces cereisiae, the whole process has the advantages of simple detection process, high detection efficiency, high accuracy and short detection period; and the invention provides a guarantee a long term development for a microecological preparation industry.

The invention discloses yeast powder for reducing and stabilizing blood sugar as well as a preparation method and application of the yeast powder. The yeast is expressed as GLP-1, and contains a glucose tolerance factor, selenium yeast and vanadium yeast converted by inorganic chromium, inorganic selenium and inorganic vanadium. According to the yeast powder disclosed by the invention, islet cells are promoted to secrete insulin through glucagon-like peptide-1; through the glucose tolerance factor, insulin resistance is reduced and insulin sensitivity is enhanced; and through the organic selenium yeast, repairing of the islet cells is promoted. From islet cell repairing and insulin releasing to insulin sensitizing, the organic vanadium directly serves as para-insulin and comprehensively treats pathogen of type II diabetes, and an effect is much better than that of a single treatment scheme; moreover, multiple efficacy factors are prepared through single yeast powder production; the yeast powder is easy for mass production, low in cost, and more economical and applicable.

Production of non-glycosylated protein from yeast is carried out by constructing expression carrier containing N-glycoamidase and purposive protein gene from molecular biological technology, transferred into yeasts from electric transformation, expressing N- glycoamidase on yeast cell surface, excretory expressing purposive protein into culture medium, removing chain of glucoprotein excretory expressed from N- glycoamidase surface expressed and obtaining non-glycosylated protein. It has better expression protein activity and correct tertiary structure and no immunogenicity to human body.

The invention discloses a transgenic method capable of controlling ALA biosynthesis in plants and promoting growth and stress resistance of plants belonging to genetic engineering field. The method is to do the trangenosis with an Arabidopsis thaliana HemAi gene photosensitive type promoter and a saccharomyces cerevisiae hansen 5-aminolevulinic acid (ALA) synthase gene (YHemi) in the plant through a plant expression vector. The invention can promote the synthesis of chlorophyll and heme and avoid the accumulation of ALA and other porphyrin intermedium in the dark through increasing ALA synthetic amount in plants, thereby avoiding plant photobleaching phenomenon and improving tolerances of plants to low temperature, high temperature, highlight, dim light and salt stress. Therefore, the method not only does not influence normal growth of the transgenic plants, but also can raise biological yield and economical yield of plants. The method can used for herbaceous plant like Arabidopis thaliana, tobacco, tomato, rape, paddy rice, watermelon, strawberry, etc, and can also used for woody plant like apple, pear, etc.

The invention discloses recomposed yeast for asymmetrically transforming and preparing (S)-4-chlorine-3-hydroxy-ethylbutyrate, which is Saccharomyces cerevisiae for reducing enzyme gene and glucose dehydrogenase gene by the introduction of carbonyl. The invention further discloses a composition method of the recomposed yeast. The method for preparing (S)-4-chlorine-3-hydroxy-ethylbutyrate by using the recomposed yeast is that: 4-chloracetyl-ethyl-acetate is used as a substrate, glucose is used as an auxiliary substrate, and the recomposed yeast is used for the transformation reaction so as to prepare (S)-4-chlorine-3-hydroxy-ethylbutyrate. The recomposed yeast, without adding any auxiliary enzyme, can effectively catalyze 4-chloracetyl-ethyl-acetate into (S)-4-chlorine-3-hydroxy-ethylbutyrate, with optical purity e.e value larger than 98 percent and the transformation ratio to the substrate larger than 95 percent, thus reducing the production cost. Compared with other strains without the need to add expensive auxiliary enzyme, the recomposed yeast has high transformation ratio to the substrate and high optical activity.

The invention discloses an easily digestible feed for sow in a lactation period. The feed is prepared from the following raw materials in parts by weight: 60-70 parts of peanut pulp, 60-70 parts of palm kernel pulp, 20-25 parts of corn fibers, 20-25 parts of green bean starch, 1-2 parts of maca, 2-3 parts of ramie roots, 3-4 parts of hemerocallis fulva, 10-13 parts of figs, 10-13 parts of malts, 20-25 parts of pennisetum purpureum schum., 8-12 parts of pennisetum sinese roxb, 10-14 parts of soy protein powder, 3-4 parts of yeast, 3-4 parts of a phagostimulant and a proper amount of water. The easily digestible feed for sow in the lactation period disclosed by the invention is prepared by a series of processes such as fermentation and puffing by taking peanut pulp, palm kernel pulp, corn fibers and green bean starch as primary raw materials. The feed is free of antibiotics, high in safety and absolutely safe and nutritional to sows in the lactation period. The feed not only is tasty and easily digestible, but also can be used for enhancing the immunity of the sow, so that the feed is a green and health nutritional feed.