82results about How to "Low nutritional requirements" patented technology

The invention discloses a pig small intestine epithelial cell line (ZYM-SIEC02), which was preserved in China Center for Type Culture Collection on January 13th, 2010 with a collection number of CCTCC-C201001. The invention also discloses the construction method of the cell line. The cell line has the advantages of low nutrient condition requirements, quick cell growth, easy cultivation and convenient promotion and application.

The invention discloses a method for producing glucaric acid by constructing recombinant yeast fermentation and belongs to the metabolic engineering field. A generation path of glucuronic acid is achieved by expressing an inositol-1-phosphate synthase gene (Inol) and an inositol oxygenase gene (MIOX) and meanwhile, aldehyde dehydrogenase (Udh) which is originated from Pseudomonas putida is expressed, so that a synthetic path of glucaric acid is successfully constructed. Recombinant pichia pastoris is used for generating 40mg / L glucaric acid in a YPD fermentation culture medium and 60mg / L glucaric acid in a YPD-MI (60mM inositol is added into the YPD culture medium) fermentation culture medium. The method for producing glucaric acid by constructing recombinant yeast fermentation has a broad development prospect, thereby laying a foundation for biosynthesis of glucaric acid.

A process for preparing the antigen protein used for hog cholera vaccine includes extracting hog cholera virus RNA, reverse transcription to obtain the immune gene E2 of hog cholera virus, using E2 as template for PCR while inserting them to expression carrier of Bichia yeast, introducing the recombinant expression carrier to Bichia yeast, screening the recombinant Bichia yeast, discriminating its expression product testing its immunoactivity, determining the chosen recombinant yeast, testing and analyzing its culture condition, choosing the optimal culture condition, culturing the recombinant yeast and preparing the antigen protein of said vaccine.

The invention relates to bacillus velezensis and an application thereof in prevention and treatment of porcine diarrhea. The invention provides a bacillus velezensis strain LL2018013. The bacillus velezensis is added into a weaned piglet milk replacer, wherein the addition amount is 107 to 109 CFU / L; and the bacillus velezensis is added into a feed for pigs at growth periods, wherein the additionamount is 106 to 108 CFU / g. The bacillus velezensis provided by the invention can improve the diversity of pig intestinal flora, reduces inflammatory factors through regulation of the flora abundance,promotes nutrient digestion and absorption, improves oxidation resistance of a body, improves immunity, increases the resisting capability to viruses, reduces generation of diarrhea, can inhibit common pig intestinal pathogens like escherichia coli K88, salmonella typhimurium and staphylococcus aureus, reduces the rate of diarrhea, improves the daily gain of pigs, can inhibit molds in a feed, improves the metabolic efficiency of the body to lipids, improves digestion and absorption of the pigs to feed nutritional ingredients, and has the trend of reducing chemical residues in excrement at thesame time.

The invention discloses an immortalized canine adipic mesenchymal stem cell line and a constructing method thereof. The immortalized canine adipic mesenchymal stem cell line capable of expressing SV40 large T antigen genes and having multidirectional differentiation is obtained by: transfecting a lentiviral vector carrying SV40 large T antigen (Ttag) by using canine adipic mesenchymal stem cells as host cells, integrating the Tag genes into an adipic mesenchymal stem cell genome and carrying out continuous passage and screening. This cell line is still active in case of 50 in-vitro passages, is high in differentiation and proliferation performance, never ages or dies, can also save medium cost and is an immortalized cell line.

The application of whterot fungus preparation in improving the continuous cropping performance of soil belongs to the field of microbe applying technology. Its implementation includes the following steps: applying whterot fungus preparation in the amount of 400-500 g / sq m into soil in the depth of 10-30 cm in two weeks before seeding, loosening soil, watering and maintaining moisture. The present invention can improve the continuous cropping performance of soil, promote the amplification of beneficial flora, preventing and controlling diseases and pests of crop, and raise the yield and quality of crop.

The invention relates to a sphingosine monad DX-T3-03 strain and an extracting method thereof. The sphingosine monad DX-T3-03 strain has high resistance against zinc and phenol degradation capability, is separated and extracted from the soil of a tailing dam of a mine, achieves high tolerance to the zinc at 4 mM / L and high degradation rate of phenol at 88 percent, has low requirements on nutrition and important application potential in biologically treating double pollution of phenol-containing industrial waste water and heavy metals; in addition, the extracting method is simple, convenient, fast, low in cost and environmental-friendly.

The invention discloses a Paecilomyces thermophila mutant strain and a mutation induction method and application thereof. The mutation induction method includes: Paecilomyces thermophila screened from high-temperature-fermented compost and used for producing high-temperature-resistant and acid-resistant glucose oxidase is used as the original strain, protoplast ultraviolet and ethyl methane sulfonate compound mutation induction is performed, and the screened strain is subjected to budding spore diethyl sulfate-microwave compound mutation induction to obtain the Paecilomyces thermophila mutant strain. The Paecilomyces thermophila mutant strain is preserved in China General Microbiological Culture Collection Center on January 11th, 2016, and the preservation number of the mutant strain is CGMCC No. 13182. Compared with a conventional mutation induction method, the mutation induction method has the advantages that the method is high in efficiency and simple in fast in screening, the productivity of the mutant strain is high, the yield of the mutant stain is 277.65% of that of the Paecilomyces thermophila original strain, fermentation liquor enzyme activity reaches 185U / ml, production cost is lowered, and the glucose oxidase produced by the mutant strain is good in high-temperature resistance and acid resistance and can be used as feed enzyme applied to feed industry.

The invention discloses a method for establishing recombinant yeast for biologically synthesizing glucuronic acid, belonging to the field of metabolic engineering. According to the method, inositol oxygenase genes (MIOX) of different resources are expressed in different yeast hosts, and biological synthesis of glucuronic acid is achieved by taking glucose or inositol as a substrate. 20mg / L of glucuronic acid is generated from a recombinant pichia pastoris strain in a YPD fermentation culture medium, and 50mg / L of pichia pastoris is generated from a YPD-MI (60mM of inositol is added into the YPD culture medium) fermentation culture medium. The method for establishing recombinant yeast for synthesizing glucuronic acid has wide development prospect, and a novel idea is provided for synthesizing glucuronic acid by using a biological method.

The invention relates to bacillus subtilis and a culture method and application thereof. The bacillus subtilis ASD05 is preserved in the General Microbiology Center of the China Committee for CultureCollection of Microorganisms on May 15, 2019, the preservation address is No.3, No.1 Courtyard, Beichen West Road, Chaoyang District, Beijing City, and the preservation number is CGMCC No.17807. The invention further relates to the culture method and application of the bacillus subtilis ASD05. It is found for the first time that the bacillus subtilis ASD05 has the effect of adhering microorganismsrich in straw degradation effect to the surface of straw, and meanwhile has the capacity of high yield of xylanase and pectinase, cellulase activity is low, and laccase activity is avoided.

The invention relates to a bacillus subtilis microbial air disinfectant for a livestock and poultry farm. The bacillus subtilis microbial air disinfectant for the livestock and poultry farm is prepared by the following steps: (1) inoculating bacillus subtilis into a liquid culture medium and performing activation culture until the logarithmic phase to prepare activated bacteria; (2) inoculating the activated bacteria into a liquid fermentation culture medium and performing fermentation culture to prepare microbial fermentation liquid; and (3) spray-drying the microbial fermentation liquid to prepare the microbial disinfectant. The bacillus subtilis microbial air disinfectant serving as spray for the first time is applied to the livestock and poultry farm and achieves antagonistic, inhibiting or killing effect on pathogenic bacteria by utilizing the growth advantage and the metabolite of the bacillus subtilis so as to disinfect and purify the farm environment.

The invention discloses bacillus coagulans H-1 and a method using the bacillus coagulans H-1 for calcium lactate production by in-situ product separating and fermentation. The strain is Bacillus coagulans H-1, and the accession number is CCTCC (China Center For Type Culture Collection) NO:M2013105. Calcium lactate with a high optical purity is obtained by seed solution culture and then 50 to 250 hours of fermentation at 50-60 DEG C. Crystallized calcium lactate crystals can be obtained by continuous fermentation with an in-situ product separating and fermentation technology until the final production concentration in a fermentation tank is 167-171g / L, the production rate can be up to 5g / L / h, the optical purity can be up to 99.7%, and the conversion rate can be up to 0.95g / g. Semi continuous fermentation process can be repeatedly combined with the in-situ product separating and fermentation technology according to needs, the separation process is simple, economic, efficient, environmental-friendly, and sustainable. According to the method for calcium lactate production by in-situ product separating and fermentation, the cost is saved, meanwhile the production efficiency is improved, and the method has important industrial application value.

The invention discloses a recombinant Escherichia coli, a construction method thereof and a method for producing glucaric acid by virtue of metabolic engineering and belongs to the field of metabolic engineering. According to the invention, a production way of glucuronic acid is realized by expressing an inositol-1-phosphate synthase gene (Ino1) and an inositol oxidase gene (MIOX) which are sourced from plants in Escherichia coli, and a synthesis route of the glucaric acid is also successfully constructed by expressing an aldehyde acid dehydrogenase gene (Udh) sourced from Agrobacterium tumefaciens, wherein the recombinant Escherichia coli generates 2.53g / L of glucaric acid in an LB-G (10g / L of glucose is added into an LB culture medium) fermentation culture medium. The method provided by the invention for producing the glucaric acid by the recombinant Escherichia coli through fermentation has a broad development prospect and lays a foundation for biosynthesis of the glucaric acid.

The invention discloses a method for constructing and differentiating a pancreatic stem cell line from human insulin. The pancreatic stem cell line is a pancreatic stem cell amplified from human insulin mass and has a multi-lineage differentiation potential. The induced pancreatic stem cell line can be differentiated to form nerve-like cells, fat-like cells, bone-like cells and functional insulin-like cell mass. The functional insulin-like cell mass is transplanted to a body of a nude mouse model suffered from diabetes, so that the blood sugar concentration of the nude mouse model can be reduced and normal blood sugar level can be maintained within one month.

The invention aims at providing a method for synthesizing alpha-ketoglutaric acid by a biological conversion method. According to the method, L-glutamic acid is used as a substrate; in a conversion culture solution obtained from kluyveromyces marxianus ATCC36534 through being cultured via a conversion culture substrate, alpha-ketoglutaricdehydrogenase inhibitors are added; the synthesis conversion culture is performed under the oscillation conditions of 25 to 30 DEG C and 200 to 250 r / min; after the synthesis conversion culture is completed, the alpha-ketoglutaric acid is obtained through separating and purifying the synthesis conversion solution; the L-glutamic acid is converted into alpha-ketoglutaric acid by yeast cells in the growth state; the alpha-ketoglutaricdehydrogenase inhibitors are added; the further metabolism consumption is blocked, so that a great amount of alpha-ketoglutaric acid is accumulated in the culture medium; when the feeing concentration of the substrate L-glutamic acid is 50g / L, the mol conversion rate can reach 83.2 percent. The low-value L-glutamic acid is converted into high-value alpha-ketoglutaric acid; the large-scale industrial application can be realized; a production process has the advantages of short period, high conversion rate, small environment pollution and the like.

The invention aims to quantitatively detect chloramphenicol by utilizing photobacterium leiognathus (YL). In the invention, the photobacterium leiognathus (YL) from marine environment is separated andpurified, and the photobacterium leiognathus is a gram-negative bacillus. China Center for Type Culture Collection (CCTCC) is authorized to preserve the photobacterium leiognathus strain on December27, 2006 with a preservation number of M 206139. The 16S rDNA gene sequence of the photobacterium leiognathus strain is submitted to the CenBank database with an access number of EF017227. The invention establishes a method system for quantitatively detecting the chloramphenicol by utilizing the characteristics of quick growth, stable luminous performance, low requirement on nutrition and sensitivity to the chloramphenicol of the photobacterium leiognathus strain and the characteristic that the concentration of the chloramphenicol is in a direct proportion to the luminous intensity suppressionratio of the photobacterium leiognathus strain through selecting indexes of an optimal detection wavelength, an optimal growth state, initiative luminous intensity, reaction time, reaction temperature, and the like so as to realize the quantitative detection of the chloramphenicol. The requirement on the quick quantitative detection of the chloramphenicol can be met by utilizing the characteristics of low cost, simple operation, high sensitivity, and the like for detecting the chloramphenicol of the invention.

The invention discloses a method for establishing goose embryo epithelial cell line and the established goose embryo epithelial cell line. The invention relates to the method for establishing the goose embryo epithelial cell line, which is characterized in that a primary goose tissue adherent method, a differential velocity enzymatic digestion and a monoclonal screening method are combined, so that primary culture condition can be optimized. The method has the advantages of simple operation process, and convenient popularization and application. The invention also relates to the goose embryo epithelial cell line established by the method, a preservation number of the goose embryo epithelial cell line is CCTCC NO: C2014137, The establishment of the goose embryo epithelial cell line solves the problem of no well-established goose source cell line in prior art. The invention also relates to a kit used for culturing and / or proliferating goose parvovirus, muscovy duck parvovirus and type I duck hepatitis virus and novel duck hepatitis virus, and is characterized in that a host cell to be infected is / or comprises the goose embryo epithelial cell line. The method for establishing the goose embryo epithelial cell line verifies the sensitive characteristic of the goose parvovirus, muscovy duck parvovirus and type I duck hepatitis virus and novel duck hepatitis virus.

The invention discloses a Trichoderma atroviride strain capable of producing high-temperature-resistant feruloyl esterase and high-temperature-resistant cellulase and application thereof. The collection number of the Trichoderma atroviride AWS26 strain is CGMCC No.8673. The Trichoderma atroviride strain has the advantages of low nutritional requirements, high reproduction speed and short fermentation period, and the fermentation products can be easily extracted and separated. The Trichoderma atroviride strain can be fermented to obtain the feruloyl esterase-cellulase composite enzyme. The composite enzyme has the advantages of high temperature resistance and high acid / alkali stability.

The invention discloses a method for degrading plant lignin by using Bacillus subtilis engineering bacterial, and belongs to the technical field of biology. According to the method, manganese peroxidase gene, lignin peroxidase gene and laccase gene are respectively cloned from microorganisms, a Bacillus subtilis expression vector containing three gene expression cassettes is constructed, the Bacillus subtilis expression vector is transformed into Bacillus subtilis, G418 resistance screening is performed to obtain the Bacillus subtilis engineering bacterial containing the manganese peroxidase gene expression cassette, the lignin peroxidase gene expression cassette and the laccase gene expression cassette, and the Bacillus subtilis engineering bacterial acts with a plant raw material to degrade plant lignin. The method has the following advantage that: the Bacillus subtilis engineering bacterial provides a stronger lignin degradation effect compared with nature microorganisms for plant lignin degradation, wherein the nature microorganisms express and secrete manganese peroxidase, lignin peroxidase and laccase.

The invention discloses a method of degrading lignin of plants with engineering bacteria of brewing yeast and belongs to the biotechnology field. The method includes following steps: (I) cloning lignin peroxidase gene, manganese peroxidase gene and laccase gene from the microorganisms; (II) constructing a brewing yeast expression carrier containing the three gene expression frames; (III) converting the brewing yeast expression carrier into brewing yeast; and (IV) performing G418 resistance screening to obtain the engineering bacteria of brewing yeast containing the gene expression frames of the lignin peroxidase gene, the manganese peroxidase gene and the laccase gene; and (V) degrading the lignin of plants by means of reaction between the three engineering bacteria of brewing yeast and plant raw materials. The degradation effect of the engineering bacteria of brewing yeast on the lignin is stronger than that of a natural microorganism which naturally expresses and secretes manganese peroxidase, lignin peroxidase and laccase and is used for degrading the lignin of the plants.

The invention aims at providing a Photobacterium leiognathi YL bacterial strain deriving from marine environment and quantitative detection on environmental pollutants by using the bacterial strain. In the invention, the luminous bacterial YL deriving from the marine environment is obtained by separating and purifying, is Gram-negative bacilli, and is rhabditiform. The luminous bacterial YL is identified to be Photobacterium leiognathi according to a phylogenetic tree established by a 16SrDNA sequencing result and a physiology and biochemistry result. The Photobacterium leiognathi YL is preserved in China Center for Type Culture Collection (CCTCC in short) with a collection number of M 206139. The 16SrDNA gene sequence of the luminous bacterial is submitted to GenBank nucleotide sequence database with the accession number of EF017227. The bacterial strain has the characteristics of rapid growth, stable luminous property, low requirements for nutrients, and sensibility to the environmental pollutants, and the like; the luminous intensity thereof is confirmed to form a maximum peak at 474 nm by fluorescence scanning; and the luminous intensity inhibition ratio thereof is in direct ratio to the concentration of multiple tested environmental pollutants, thereby the quantitative detection of the tested environmental pollutants can be realized. The invention has the characteristics of low cost, simple and convenient operation, high sensitivity and the like on detecting the environmental pollutants, , thereby satisfying the requirements on rapid detection of the pollutants.

The invention discloses a preparation method of a bacterial cell catalyst for catalyzing naringin ester synthesis reaction. The method comprises the following steps: inoculating a bacterium seed solution into an enzyme-inducer-containing liquid culture medium, and culturing at 30-50 DEG C for 12-144 hours, wherein the inoculum size is 0.5-20% (v / v); and centrifuging, collecting the thallus, washing with distilled water twice, carrying out vacuum freeze drying for 12-36 hours, and collecting the thallus dry powder to obtain the bacterial cell catalyst. The catalyst is used for catalyzing ester conversion reaction between esculine and an acyl reagent at high selectivity. Compared with the enzyme preparation, the obtained bacterial full-cell catalyst avoids the expensive and complex enzyme separation purification process, is easy to prepare and recover, is more beneficial to large-scale production, and has higher stability in organic solvents.

The invention provides photobacterium leiognathi (Photobacterium leiognathi YL) from a marine environment and quantitative detection of environmental pollutants by using the strain. A strain of luminous bacterium YL from the marine environment is separated and purified, and the strain is Gram-negative bacterium and has a rod shape. The luminous bacterium YL is identified as the Photobacterium leiognathi according to a phylogenesis tree constructed according to a 16S rDNA sequencing result and physiological and biochemical results. The strain was preserved in China Center for Type Culture Collection (CCTCC) on 27, December, 2006, and the collection number is M 206139. The 16S rDNA gene sequence of the luminous bacterium has been submitted to a GenBank nucleotide sequence database, and the access number is EF017227. The strain has the characteristics of fast growth, stable luminous performance, low nutrition requirement, sensitivity to environmental pollutants, and the like, and the luminous intensity of the luminous bacterium is determined to form the maximum peak at 474nm through fluorescent scanning. The luminous intensity inhibition rate is in a direct proportion to the concentration of various environmental pollutants, and the quantitative detection of the environmental pollutants can be realized. The method for detecting the environmental pollutants by the strain has the characteristics that: the cost is low and the method is easy and convenient to operate, high in sensitivity and the like, and can meet the requirement on rapid detection of the pollutants.

The invention discloses a method for constructing a human islet-derived pancreatic stem cell line and a method for differentiation of the human islet-derived pancreatic stem cell line into insulin-producing cells. The pancreatic stem cell line is obtained by amplifying pancreatic stem cells in human pancreatic clusters, and is a potential source for multidirectional differentiation. When induced, the pancreatic stem cell line can differentiate to form neuron-like cells, adipose-like cells, osteoblast-like cells and functional islet-like cell clusters. By transplanting the functional islet-like cell clusters into a body of a diabetic nude mouse model, the blood glucose concentration of the nude mouse model can be reduced and reach a normal level within 24 days.

The invention discloses a fermentation preparation method, and a separation and purification method of heat-resistant and acid-resistant glucose oxidase, and belongs to the technical field of microorganisms. The glucose oxidase prepared by fermentation has high heat resistance and acid resistance, and the defect of low enzyme activity caused by use in food and feed in a high temperature and acidic environment is overcome. The fermentation preparation method comprises the following specific steps: culturing a bacterial strain into a spore, cleaning the spore and then preparing a mono-spore suspension; introducing the mono-spore suspension into a seed culture medium for fermentation culture; introducing seed culture fluid into a shaking flask fermentation medium for fermentation culture, adding a fermentation supplementary nutrient solution into fermentation liquid; adding dextrin, soluble starch and potassium sorbate into the fermentation liquid, and uniformly stirring; freezing, drying and crushing the fermentation liquid to obtain the glucose oxidase. After crude enzyme fluid of the glucose oxidase obtained by the fermentation is subjected to super-heating treatment, ammonium sulfate-ethanol-compound precipitation, ion exchange chromatography and molecular sieve gel filtration chromatography, the glucose oxidase with high recovery rate and high purity is obtained.

The invention relates to penicillium citrinum LB and application in forsythiaside preparation through biotransformation of forsythin. The application comprises the following steps: with the wet thalli obtained by fermentation culture of penicillium citrinum LB as a biocatalyst, forsythin as a substrate and methanol as a cosolvent, forming a transformation system from fermentation liquid containing the wet thalli or by suspending the wet thalli in a buffer solution; performing a transformation reaction under a constant-temperature oscillation condition at 30-35 DEG C and at a speed of 200-250r / min; and after the transformation reaction, separating and purifying the transformed liquid to obtain forsythiaside. When the feed concentration of the substrate is 2g / L, the transformation yield of forsythiaside is 90.5%. The penicillium citrinum LB has the advantages of low nutrition requirements, short fermentation time, strong anti-pollution ability, fast growth, good specificity of transformation reaction, high transformation efficiency and few side products.

The invention provides a heat resistant beta-amylase-trehalose synthase fusion enzyme, an expression gene of the heat resistant beta-amylase-trehalose synthase fusion enzyme, an engineering bacterium secreting the fusion enzyme and application, and belongs to the technical field of genetic engineering. The heat resistant beta-amylase-trehalose synthase fusion enzyme comprises an amino acid sequence with SEQ ID No.1 and another amino acid sequence, and the similarity of the another amino acid sequence to the amino acid sequence with SEQ ID No.1 is 80%-100%. The expression gene of the heat resistant beta-amylase-trehalose synthase fusion enzyme comprises an amino acid sequence with SEQ ID No.2 and another amino acid sequence, and the similarity of the another amino acid sequence to the amino acid sequence with SEQ ID No.2 is 80%-100%. The obtained beta-amylase-trehalose synthase fusion enzyme has a heat resistant property, directly converts amylose into trehalose at the higher temperature, saves the complicated steps of separating and purifying endoenzyme, simplifies the production process of the trehalose, and reduces the production cost.

The invention discloses a preparation method of a mould cell catalyst for catalyzing synthetic reaction of cytarabine ocfosfate. The method comprises the following steps of: inoculating mould spores into a liquid basal culture medium containing an enzyme production inducer with the concentration of 0.5g / L-50g / L, wherein the inoculum concentration is 1*103-1*1012 mould spores / L, the culture condition is 10 DEG C-45 DEG C and the culture time is 10 hours-200 hours; collecting thalli, washing with distilled water or a buffer solution, then washing with acetone for 1-3 times, and performing suction filtration on the thalli to obtain the mould cell catalyst for catalyzing synthetic reaction of cytarabine ocfosfate; or removing water by a lyophilizing method to obtain the mould cell catalyst for catalyzing synthetic reaction of cytarabine ocfosfate. The mould cell catalyst disclosed by the invention has the advantages of low cost, easiness of preparation, environmental friendliness and high catalyst activity.

The invention belongs to the technical field of industrial microorganism, and in particular relates to fungus capable of efficiently degrading multiple types of dyes under non-sterilization condition. Microorganism from the soil of a phyllostachys praecox forest in Hangzhou Lin'an is separated on a PDA culture medium by a gradient dilution method and is subjected to laboratory screening, so that the important fungus-trichaptum abietinum 1302BG capable of efficiently degrading multiple types of dyes under the non-sterilization condition is obtained. The strain can decolorize multiple types of dyes, has wide pH adaptability, has low requirement on nutrient components of the culture medium, and still has efficient decolorization under the non-sterilization condition even the condition of artificially added with certain other microorganism pollution source. Dye solution decolorized by the strain does not have toxicity or the toxicity is largely reduced. The excellent characteristics show that the strain has high treatment and application capability for industrial dye wastewater.

The invention provides an ecological reconstruction method for a concrete spray layer surface based on growth of moss. According to the method, wild natural growth drought-enduring moss varieties are selected, after the moss is dried in the shade and cut, the moss is mixed with other auxiliary materials according to a corresponding proportion to prepare a medium for the growth of the moss, the medium is directly laid on a slope of the concrete spray layer, a corresponding maintenance measure is taken, and then ecological greening of the concrete spray layer is achieved. According to the ecological reconstruction method for the concrete spray layer surface based on the growth of the moss, the problem that plants cannot grow on the surface of the concrete spray layer, and thus greening is seriously lacked can be effectively solved, a new means is opened up for erosion control and ecological green returning, and the industrial prospect is very wide.