62 results about "Aspergillus usamii" patented technology

The invention provides rapeseed protein feed which is solid powder or particulate matter. The content of protein is between 35 and 50 percent (N * 6.25, dry basis); the content of water-soluble nitrogen is between 25 and 35 percent; the content of small peptide is between 10 and 25 percent; the content is counted by weight percentage; and thioglycoside accounts for 5 to 15 micromole per gram of rapeseed protein feed. The invention also provides a method for preparing the rapeseed protein feed; one of bacillus subtilis, lactobacillus or candida utilis and actinomucor elegans or aspergillus usamii are compounded to prepare a fermenting agent; the fermenting agent is added to pulverized rapeseed dreg for solid fermentation; and a culturing substrate is sterilized, dried and pulverized to obtain the rapeseed protein feed rich in small peptide. The preparation method is environment-friendly, has mild reaction condition, can fully exert the functions of protease hydrolysis and thioglycoside degradation of microbe and is a method which has low cost and is suitable for the industrialized large-batch preparation of the rapeseed protein feed.

The invention discloses a composite microbial inoculum for composting organic materials and a preparation method thereof. The microbial inoculum mainly comprises pichia pastoris, lactobacillus acidophilus, bacillus coagulans, enterococcus faecalis, bacillus natto, bacillus subtilis, aspergillus nidulans, white-rot fungi, aspergillus usamii, trichoderma harzianum, trichoderma reesei and aspergillus niger. The preparation method comprises the following steps: first classifying the microorganisms into bacteria, yeasts and filamentous fungi, then performing mixed culture on the same class of microorganisms till quality requirements are met, and then compounding according to the following proportions: 20-70 parts of bacteria, 30-60 parts of yeasts and 50-80 parts of filamentous fungi. The compositing microbial inoculum prepared by the preparation method has the advantages of fast fermentation start during composting fermentation, high temperature in the composting process, a long high-temperature period and fast organic material decomposition. An organic fertilizer prepared by using the microbial inoculum is good in quality; compost has a fertility-retaining function, has high effective utilization rate of a large number of elements such as nitrogen, phosphorus and potassium, and has the effects of dissolving phosphorus, dissolving potassium and fixing nitrogen; compared with a control group which is not inoculated with the microbial inoculum, a product provided by the invention has the advantages as follows: the content of water-soluble organic carbon is increased by about 2 times, the total humic acid content is increased by about 1 times or above, and the content of biochemical fulvic acid is increased by about 2 times.

The invention discloses aspergillus usamii epoxide hydrolase mutants with improved enantioselectivity, and belongs to the technical field of enzyme engineering and biological catalysis. Aspergillus usamii epoxide hydrolases (AuEH2) are subjected to molecular modification on the basis of rational designs and are combined with site-saturation mutagenesis methods for genes, so that the multiple epoxide hydrolase mutants with the improved enantioselectivity can be obtained. The aspergillus usamii epoxide hydrolase mutants have the advantages that racemic styrene oxide (rac-SO) can be catalyzed by the six mutants AuEH2A250I, AuEH2A250M, AuEH2A250Y, AuEH2A250S, AuEH2A250L and AuEH2A250V with the improved enantioselectivity, and the enantiomeric ratios (E values) of the six mutants can be increased and respectively reach 48, 27.2, 23.9, 21.8, 20.0 and 19.4 from the original 16 as compared with wild type mutants.

The invention provides a method for preparing rapeseed active peptide by using mixing bacteria to perform solid state fermentation. The method comprises the following steps: rapeseed pulp obtained by pressing or leaching oil preparation process is crushed to obtain crushed rapeseed pulp; one of bacillus subtilis, lactic acid bacteria or candida utilis is compounded with actinomucor elegans or aspergillus usamii to prepare a fermenting agent; the fermenting agent is injected in the crushed rapeseed pulp for solid state fermentation; a culture medium after solid state fermentation is extracted by adding water, and the residue is removed by separation so as to obtain a coarse extracting solution of the rapeseed active peptide; and the coarse extracting solution is decolored, desalted and dried to obtain the rapeseed active peptide. The method has the advantages of environmental protection and mild reaction conditions, and can fully utilize hydrolytic action and sulfur glycoside degradation action of protease, so the method is low in cost and suitable for industrialized production of the rapeseed active peptide.

The industrial process of producing xylanase with Aspergillus usamii belongs to the field of enzyme engineering technology. In the industrial process of producing xylanase, bagasse, corn cob, bran and other crop leftovers are used as the fermenting base material, and proper amount of inorganic salt, surfactant, water, etc are added for solid fermentation in a 30M3 solid fermenting tank with Aspergillus usamii E001 strain. The mature fermented material is stoved with hot blast at 45-50 deg.c to obtain xylanase product possessing xylanase activity of 7783-9616 IU / g. The present invention provides also the recipe of fermenting culture medium and the fermenting conditions.

The invention provides a method for preparing an organic zinc enzyme preparation. The method comprises the following steps of: mixing materials such as bran, bean pulp, powdered rice hulls, solution of zinc sulfate and the like to obtain a mixture, stirring uniformly, then performing high-temperature high-pressure sterilization, inoculating a koji plate strain of Candida utilis after the sterilization, stirring uniformly and then culturing, then taking a cultured material out, inoculating a koji plate strain of Aspergillus usamii, culturing again, taking a subcultured material out, inoculating a koji plate strain of bacillus subtilis, stirring uniformly and then culturing for the third time, and oven-drying and crushing a material obtained by the culturing for the third time to obtain the organic zinc enzyme preparation. In the method, a production technology is simple, the production cost is low, the three bacteria are adopted for three continuous transformations, and the transformation ratio from inorganic zinc to organic zinc is up to over 98 percent. The prepared enzyme preparation integrates organic trace elements, an enzyme preparation and probiotics into a whole, and can increase the disease resistance of raised animals and promote the healthy growth of the raised animals.

The invention aims at providing a method for thermal stability modification of GH10 xylanase Aus Xyn10A and efficient expression and purification of recombinant mutant enzyme. According to the protein sequence comparison result between Aus Xyn10A (from aspergillus usamii E001) and heat-resistant xylanase (from thermoascus aurantiacus 751K6A), the sequence of the N-terminal area of the heat-resistant enzyme is replaced by a corresponding sequence of Aus Xyn10A by a genetic engineering method. The obtained mutant enzyme is named ATx10AM. The experimental result indicates that the optimal temperature and thermal stability of the enzyme after mutation are obviously improved; and as a heat-resistant enzyme preparation, the xylanase has relatively great industrial production potential and economic value.

The invention aims to provide a high-efficiency expression and purification method for family-10 xylanase Aus Xyn10A thermostability modified and reconstructed mutant enzyme. According to a comparison result of protein sequences of the Aus Xyn10A (derived from Aspergillus usamii E001) and thermostable xylanase (derived from Thermotoga maritima), the C terminal repeat of thermostable enzyme is used to the C terminal of the Aus Xyn10A by a gene engineering method, and the obtained mutant enzyme is named Aus Xyn10A'. Experimental results prove that the optimum temperature of the mutant enzyme is obviously improved, and the thermostable enzyme serving as a thermostable enzyme preparation has high industrial production potential and economic value.

The method relates to a method for preparing a sumac extract by adopting aspergillus fermentation of aspergillus oryzae and the like and the sumac extract prepared by the method, in particular to the method for preparing the sumac extract with the following steps: a) the sumac is dried, grinded and water is added to carry out moisture absorption on the sumac; b) hygroscopic material prepared in step a) is cooked; c) aspergillus is inoculated, nurtured and fermented in the cooked material prepared in step b), the aspergillus is selected from a group composed of aspergillus oryzae, aspergillus kawachii, aspergillus usamii, aspergillus shirousamii, aspergillus niger and aspergillus awamori; d) water is added to leavening prepared in step c) and is heated; e) liquid ingredients are recovered from the heated material prepared in step d). The method for preparing the sumac extract of the invention sharply improves efficiency of numerous active ingredients of the sumac, eliminates toxic ingredients thereof and improves functional characteristics of the prepared sumac extract.

The invention provides a cloning method for complete mRNA and DNA sequences of beta-1,4-endo-glucanase gene from Aspergillus usamii E001. The nucleotide sequences of the complete mRNA and DNA are shown in SEQ ID NO:1 and SEQ ID NO:2 respectively. Bioinformatics analysis shows that the beta-1,4-endo-glucanase belongs to glycoside hydrolase family 12, is named Aus Cel12A, and has the amino acid sequence shown in SEQ ID NO:3; and the corresponding gene is named Aus cel12A. The invention also discloses a method for construction of Aus Cel12A engineering bacteria and high-efficiency expression and purification of recombinant Aus Cel12A. The most suitable operative temperature and pH of the prepared recombinant Aus Cel12A are respectively 60DEG C and 5.0; the recombinant Aus Cel12A is stable when the pH is 4.0-7.0 and the temperature is below 60DEG C, has higher industrial production potential and economic value, and lays a theoretical basis for researching other beta-endo-glucanase.

The invention provides a preparation method of an organic iron enzyme preparation, which comprises the steps of: uniformly mixing raw materials, such as wheat bran, bean pulp, rice husk powder, a ferrous sulfate solution and the like to prepare a mixture, carrying out high-temperature high-pressure sterilization, inoculating a koji tray strain of candida utilis after sterilizing, uniformly stirring and then culturing, then taking materials obtained by the culturing out, inoculating a koji tray strain of aspergillus usamii, uniformly stirring and then subculturing, then taking materials obtained by the subculturing out, inoculating a koji tray strain of bacillus subtilis, uniformly stirring and then culturing for the third time, and drying and crushing materials obtained by the third-time culturing to prepare the organic iron enzyme preparation. The method has the advantages of simple preparation process and low production cost; three times of transformation are continually carried out by adopting the three stains, and inorganic iron is transformed into organic iron with high transformation rate reaching above 98 percent; and the prepared enzyme preparation integrates organic trace elements, an enzyme preparation and probiotics into a whole, has multiple biological functions, and can enhance the disease-resistant capacity and the stress-resisting function on a fed animal.

The invention discloses a method for preparing (S)-styrene oxide through an enzyme method, which belongs to the technical field of biological catalysis. According to the method provided by the invention, in a two-phase system of hexyl alcohol / buffer solution, epoxide hydrolases from aspergillus usamii is utilized to catalyze hydrolytic kinetic resolution of racemization styrene oxide to prepare the (S)-styrene oxide. Compared with a single-phase reaction system, the concentration of the kinetic resolution rac-SO is improved to 120g / L from 24g / L; the space time yield is improved to 20.3g / L / h from 3.1g / L / h; an ee value for preparing the (S)-SO by kinetic resolution 120g / L rac-SO is improved to 98.3 percent from 36.8 percent, and the gram-scale preparation of the (S)-SO is realized. The method provided by the invention is simple in process, a product is high in enantiomer purity and yield, high in catalytic efficiency and environmental friendly, and the method has wider industrial application prospect.

The invention discloses a method for producing novel haematochrome from a pseudo-ginseng residue. The method is realized by the following steps: (1) carrying out pretreatments including drying, crushing and sieving on the pseudo-ginseng residue; (2) uniformly mixing the pretreated pseudo-ginseng residue with water and a nitrogen source in a certain proportion; (3) carrying out steam sterilization on the uniformly mixed material; (4) inoculating the sterilized material with at least one monascus and at least one of Aspergillus niger, Aspergillus usamii and rhizopus oryzae, and carrying out mixed bacteria solid state fermentation; and (5) drying the obtained fermented culture, so as to obtain the novel haematochrome. By utilizing the method, the pseudo-ginseng residue can be converted into the novel haematochrome; and the pseudo-ginseng residue is subjected to resource utilization and is then turned into wealth, so that environmental pollution caused by the pseudo-ginseng residue is reduced, and novel haematochrome is provided for the food and medicine industries.

The invention belongs to the field of feed preparation, and particularly relates to a method for preparing a feed additive from cymbopogon citratus residues. The method comprises the following steps of taking trichoderma koningii, aspergillus usamii and geotrichum candidum for fermenting cymbopogon citratus residues, star aniseed residues and Chinese cinnamon residues, putting the fermented materials to a drying device, and after the dried materials are crushed with a crusher, filtering the crushed materials with a 60-mesh sieve. According to the method for preparing the feed additive from cymbopogon citratus residues, provided by the invention, waste is sufficiently used, and when being used for breeding, the made feed has good health-care medicine effects on livestock.

The invention provides a preparation method of an organic manganese enzyme preparation, which comprises the steps of: uniformly mixing raw materials, such as wheat bran, bean pulp, rice husk powder, a manganese sulfate solution and the like to prepare a mixture, carrying out high-temperature high-pressure sterilization, inoculating a koji tray strain of candida utilis, uniformly stirring and then culturing, then taking materials obtained by the culturing out, inoculating a koji tray strain of aspergillus usamii, uniformly stirring and then subculturing, then taking materials obtained by the subculturing out, inoculating a koji tray strain of bacillus subtilis to the materials obtained by the subculturing, uniformly stirring and then culturing for the third time, and drying and crushing materials obtained by the third-time culturing to prepare the organic manganese enzyme preparation. The method has the advantages of simple preparation process and low production cost; three times of transformation are continually carried out by adopting the three stains, and inorganic manganese is transformed into organic manganese with high transformation rate reaching above 98 percent; and the prepared enzyme preparation integrates organic trace elements, an enzyme preparation and probiotics into a whole and has multiple biological functions, simple production process and low production cost.

The invention provides a building method of a novel pichia pastoris coexpression system for co-expressing engineering bacteria GS115 / faeA-xyn11A originated from aspergillus usamii E001 feruloyl esterase A gene and beta-1, 4-xylanase gene. The built pichia pastoris coexpression system can be used for the industrial production of the feruloyl esterase and the beta-1, 4-xylanase. The prepared two enzymes have the characteristics of being high in substrate affinity and high in catalytic efficiency, and are higher in industrial production potential and economic value.

The invention relates to the field of microbial fermentation, and in particular to a method for fermentation production of beta-mannase. The method comprises the step of expressing beta-mannase by aspergillus usamii By247 through fermentation. According to the mannose produced by the invention, palm meal is used as one of the fermenting materials, and animal experimental results show that the mannase can decompose antinutritional factors in the palm meal in unconventional feed well and remarkably improve the production performance of animals. The temperature resistance, the feed and material decomposing capacity of the mannase and the result of animal experiments are better than those of other merchant beta-mannase.

The invention discloses a method for preparing (R)-phenylglycol (PED) from racemic styrene oxide (rac-SO) under catalytic actions of double enzymes, belonging to the technical field of biological catalysis. By using rac-SO as the substrate, double epoxide hydrolases (epoxide hydrolase mutant AuEH2A250I and epoxide hydrolase VrEH3) are utilized to perform enantioconvergent hydrolysis on the rac-SO, thereby preparing the high-enantiopurity product (R)-PED. The VrEH3 derived from mung beans and AuEH2A250I derived from Aspergillus usamii are combined to perform enantioconvergent hydrolysis on the rac-SO for the first time in the invention. The ee value for preparing (R)-PED by using catalyzing 10mM rac-SO is 96.0%. The method has ideal behaviors applicable to industrial application, lays theoretical foundation for industrialized production of enzymes, and has higher industrialized application potential and economic value.

The invention provides a method for cloning and analyzing complete micro ribonucleic acid (mRNA) and deoxyribonucleic acid (DNA) sequences of a novel endo-beta-1,4-glucanase gene derived from an aspergillus usamii E001 strain. Nucleotide sequences of the complete mRNA and DNA sequences are shown as SEQ ID NO. 1 and SEQ ID NO. 2 respectively. Bioinformatic analysis shows that the endo-beta-1,4-glucanase belongs to the group V of glycoside hydrolase and is called Aus Cel5A, wherein the amino acid sequence of the glucanase is shown as SEQ ID No. 3; and a corresponding gene is called Aus cel5A. A theoretical basis is laid for heterogeneous expression and industrial production of the gene; greater industrial production and application potential and economic value are achieved; and a theoretical basis is laid for research of other endo-beta-1,4-glucanases.

The invention provides a preparation method of an organic selenium enzyme preparation, comprising the following steps of: mixing bran, bean pulp, rice hull powder, a sodium selenite solution, and the like, sterilizing at high temperature and high pressure, inoculating a kojitray strain of candida utilis after sterilization, culturing and then taking out a material obtained through culturing, inoculating a kojitray strain of aspergillus usamii on the material obtained through culturing, culturing for the second time, inoculating a kojitray strain of bacillus subtilis on a material obtained through culturing for the second time, culturing for the third time, drying and crushing a material obtained through culturing for the third time to prepare the organic selenium enzyme preparation. In the preparation method of the organic selenium enzyme preparation, continuous three times of conventions are carried out by adopting three types of strains, the conversion ratio of converting original inorganic selenium with extremely low valence into organic selenium with higher valence can reach above 98 percent, and the prepared enzyme preparation integrates organic trace elements, enzyme preparation and probiotic bacteria into a whole, simultaneously has a plurality of biological functions and has the efficacy of reducing anti-nutritional factors.

The invention relates to a method for preparing carboxypeptidase from aspergillus usamii and belongs to the technical field of biological engineering. In the method, agricultural and sideline products such as brans, bean cake powder, corn meal and the like are used as fermentation base materials; and the carboxypeptidase is prepared by suitably changing the ratio of a carbon source to a nitrogen source in a culture medium and a crude enzyme extraction time and utilizing the aspergillus usamii E001 to carry out solid fermentation. The invention provides a solid fermentation culture medium formula and a culturing condition for experimental production in a laboratory. The activity of the carboxypeptidase for maturely fermenting yeast materials reaches 3,452.4 to 5,321.6U per gram of brans. The method has the characteristics of simple equipment, low investment, high response, low production cost, environment friendliness, higher activity of the carboxypeptidase and the like, is beneficial to development and utilization of the carboxypeptidase and has higher economic and practical values.

The invention discloses mannose oligomer oat vinegar and a preparation method thereof. The problems that in existing oat vinegar, nutritional ingredients are single, the mouthfeel and health-care functions are expected to be further improved, and a large quantity of konjac flying powder, waste yeast and red algae are abandoned and not effectively utilized are solved. According to the technical scheme, the preparation method comprises the steps that mixed slurry of the konjac flying powder, the waste yeast and the red algae is hydrolyzed through beta-mannosidase generated by aspergillus usamii and then mixed with oat slurry, and amylase and glucoamylase are added to obtain saccharified liquid; in the fermentation process, saccharomyces cerevisiae, aroma-producing yeast and acetobacter pasteurianus are added for further fermentation, and then the mannose oligomer oat vinegar is prepared. The preparation method is simple and low in production cost, the konjac flying powder, the waste yeast and the red algae are effectively utilized, the prepared oat vinegar is rich in nutritional ingredient and pure in flavor, the health-care effect of the mannose oligomer oat vinegar is enhanced, and the quality and health-care effect of the product are improved.

The invention provides a cloning method of complete mRNA and DNA sequences of a novel endo-chitosanase gene originated from Aspergillus usamii E001 strain. The nucleotide sequences are respectively SEQ ID NO:1 and SEQ ID NO:2. It shows through bioinformatics analysis that the endo-chitosanase belongs to glucoside hydrolase family 75 and is named as Aus CsnA. The amino acid sequence is SEQ ID NO:3 and the corresponding gene is named as Aus csnA. The invention also discloses an Aus CsnA engineering bacteria construction method and a recombinant Aus CsnA highly expression and purification method. The most appropriate operation temperature and pH of the prepared recombinant Aus CsnA are respectively 50 DEG C and 5.0, and the recombinant Aus CsnA is stable below 50 DEG C when pH is 4.0-7.0. The invention has great industrial production potential and economical values.

The invention discloses a method for producing chitosanase by using Aspergillus usamii, which belongs to the technical field of biological engineering. In the method, agricultural and sideline products such as bran, soybean cake powder and rapeseed cake powder are used as fermentation substrates, a proper amount of inorganic nitrogen source, chitosan, inorganic salt, surfactant and the like are added, and the chitosanase is produced by the solid fermentation of an Aspergillus usamii E001 strain. The invention provides the formula and culture conditions of a solid fermentation culture medium for test production of chitosanase by the Aspergillus usamii E001 in a laboratory, and the chitosanase activity of a mature fermented grain material of the medium reaches 1,942 to 2,461U / g. The method has the advantages of simple equipment, small investment, quick effectiveness, low production cost, environmentally-friendly, high activity of chitosanase fermentation enzyme, and the like, contributes to the development and utilization of chitosanase and has high economic and practical value.

The invention relates to a method for preparing rapeseed peptides by microorganism liquid fermentation, which sufficiently utilizes the functions of hydrolyzing proteins and degrading sulfuric glucoside by microorganism, thereby effectively improving the soluble nitrogen content and the bioactivity of the product. The method for preparing the rapeseed peptides by microorganism liquid fermentationcomprises the following steps: (1) mixing rapeseed meal powder with nutrient solution to prepare liquid fermentation medium; (2) inoculating the microorganism into the fermentation medium to carry outliquid fermentation; (3) carrying out solid-liquid separation on the liquid fermentation medium after fermentation to obtain the crude extract of the rapeseed peptides; and (4) purifying the crude extract to obtain the rapeseed peptides. A strain capable of generating protease is actinomucor elegans, aspergillus usamii, bacillus subtilis, lactobacillus or candida utilis. The invention has simplepreparation process, utilizes the microorganism liquid fermentation to generate a plurality of metabolites and enzymes and sufficiently utilizes the functions of hydrolyzing proteins and degrading sulfuric glucoside by the microorganism, thereby improving the soluble nitrogen content, the bioactivity and the yield of the rapeseed peptides.

The invention provides a novel construction method of engineering bacteria GS115 / man5A-xyn II for co-expressing a beta-mannase gene derived from Aspergillus niger LW-1 and a beta-1,4-xylanase gene derived from Aspergillus usamii E001. A constructed Pichia pastoris co-expression system can be used for industrial production of beta-mannase and beta-1,4-xylanase. The prepared two enzymes have the characteristics of strong substrate appetency and high catalytic efficiency and have large industrial production potentials and economic values.

The invention discloses mold-culture-containing enzyme special for growing pigs and a preparation method thereof and belongs to the technical field of preparation of enzyme preparations. Barley malt, burnt malt, figs and pineapples which are rich in a variety of enzyme systems and garlic, vanilla beans, illicium verum and fennel which can enhance animal appetite and resist to and inhibit bacteria serve as main raw materials, the low temperature wall breaking and extraction technologies are adopted, effective constituents in the raw materials are extracted to the greatest extent, natural plant extract is prepared, meanwhile, aspergillus usamii which can produce high-vitality acid protease serves as an original strain for producing a mold culture containing high-vitality acidic protease, Chinese herbal extract, protective agents, activating agents, anti-oxidant and other food-grade feed enzyme are compounded scientifically, by means of the obtained feed compound enzyme, safe and comprehensive digestive enzyme is provided for the growing pigs, in addition, pleasant fragrance can be brought to feed, appetite of the growing pigs is enhanced, the digestion and absorption functions of the stomachs and intestines of the growing pigs are effectively improved, and productivity of the growing pigs is raised.

The invention discloses Aspergillus usamii epoxide hydrolase mutants with improved enantioselectivity, and belongs to the technical fields of enzyme engineering and biocatalysis. Based on rational design, molecular transformation of the Aspergillus usamii epoxide hydrolase (AuEH2) is carried out; and combined with a site-directed saturation mutation method, a plurality of epoxide hydrolase mutantswith enhanced enantioselectivity are obtained. Enantiomeric ratios (E value) of the 9 enantioselectivity-enhanced mutants catalyzing rac-pMPGE are increases to 32.4, 31.1, 27.8, 25.8, 23.5, 22.4, 20.2, 14.1, and 13.6 correspondingly compared to an E value of 12.7 of a wild type.

The invention relates to a microecological ferment for fattening pig feeds and a manufacturing method thereof, and an active solid-state fermentation feed. The microecological ferment is composed of the following components in parts by weight: 3-5 parts of Aspergillus usamii, 1-3 parts of Rhizopus oryzae, 1-3 parts of Debaromyces hansenii, 1-3 parts of Geotrichum candidum, 2-4 parts of Bacillus subtilis subsp.subtilis, 2-4 parts of Aspergillus niger and 1-3 parts of Mortierella minutissima. The screening and combination of a plurality of strains are performed to solve the problems of singleness, incomprehensive fermentation and poor fermentation effect in the existing microbial fermentation preparation, thereby achieving the goals of enhancing the product quality and lowering the production cost. When being used for breeding fattening pigs, the feed provided by the invention has obvious effects on promoting growth of fattening pigs and enhancing the disease resistance, has the characteristics of high feed utilization efficiency and the like, and reduces the breeding environment pollution. The bred product has the advantages of no residues of antibiotics, hormones or other drugs, abundant nutrition, and pure and natural taste.