44 results about "Rhizomucor miehei" patented technology

There is provided an enzymatic detergent composition which comprises: (a) surfactant; (b) 10-20,000 LU per gram of the detergent composition of a lipolytic enzyme obtainable from Humicola lanuginosa, Pseudomonas pseudoalcaligenes, Rhizomucor miehei and (c) a non-cross-bridged polydentate N-donor ligand capable of forming a complex with a transition metal, wherein said complex is capable of catalysing the bleaching of stains on fabrics by means of atmospheric oxygen.

The invention relates to an improved rhizomucor miehei lipase gene and an application to yeast display. The sequence of the improved rhizomucor miehei lipase gene is SEQ.ID.No2, with respect to a recombinant vector pMD18-T-RML containing the gene, RML means lipase gene; and the collection number of a bacterial strain Escherichia coli TOP10 / pMD18-T-RML carrying the plasmid is CCTCC M 208136. In the invention, the gene is transferred into pichia stipitis host strain, so that the rhizomucor miehei lipase is displayed and expressed in the pichia stipitis. The provided pichia stipitis can effectively display the rhizomucor miehei lipase. The lipase can be widely applicable for producing fatty acid methyl ester, ethyl caproate, triglycerides which have different melting points but does not contain various types of fatty acid and a few 'reconstructed esters'.

The invention discloses recombinant pichia pastoris for heterogenous high level secretory expression of a rhizomucor miehei lipase. The recombinant pichia pastoris is obtained by introducing a self leader peptide gene for encoding the rhizomucor miehei lipase by employing a molecular biology technology, then optimizing a copy number of an objective gene, and constructing a vector which can be expressed in pasteur pichia pastoris. Researches show that: the introduction of the self leader peptide greatly increases the secretion amount of the objective lipase, a pichia pastoris recombinant of a lipase gene (pro-rml) containing two copies has highest enzyme activity, and the lipase Pro-RML has higher activity of hydrolysis of tri-acylglycerol.

PCT No. PCT / DK95 / 00511 Sec. 371 Date Aug. 14, 1997 Sec. 102(e) Date Aug. 14, 1997 PCT Filed Dec. 20, 1995 PCT Pub. No. WO96 / 19582 PCT Pub. Date Jun. 27, 1996A method of producing a milk clotting enzyme including the steps of (a) fermenting a strain of Rhizomucor miehei or Aspergillus oryzae to form a fermentation product having a glycoslated Rhizomucor miehei aspartic protease and other proteins, and (b) subjecting a quantity of the fermentation product to a deglycosylating treatment to form a coagulant preparation having an at least partly deglycoslated aspartic protease and the other proteins. The at least partly deglycosylated protease has a milk clotting activity that is at least 10% higher than a milk clotting activity of the glycosylated aspartic protease.

A method for producing a lipase powder, which comprises the step of spray drying an aqueous solution containing a lipase adjusted to pH 6 to 7.5; the lipase powder which is a 1,3-specific lipase derived from Rhizomucor miehei, spherical, and contains not more than 10% by weight of moisture; and a lipase composition wherein the lipase powder is immersed or impregnated in fatty oil. According to the present invention, a lipase powder of which 1,3-selectivity is improved can be provided.

Topical compositions comprising extracts of Rhizomucor miehei that are substantially devoid of acid-protease activity and their use in treating dermatologic conditions, including reducing the appearance of signs of skin aging.

The invention discloses recombinant pichia pastoris for heterologously and efficiently expressing lipase and application of the recombinant pichia pastoris. The recombinant pichia pastoris is obtained through converting plasmid SEC31-pPIC3.5K overexpressing a gene SEC31 in recombinant pichia pastoris X-33, which contains a 4-copy number pro-rml gene and can express Pro-RML. According to the recombinant pichia pastoris and the application thereof, the expression of the rhizomucor miehei lipase is effectively promoted, the efficiency of secretion of the lipase is increased, the enzyme activity reaches 996U / mL after 144 hours of shake-flask fermentation, and the strain ectoenzyme activity secretion efficiency reaches 38U / OD600 and is higher than that (26U / OD600) of two recombinant pichia pastoris strains of rhizomucor miehei lipase copy numbers concerned in the patent CN103361327A.

The invention provides a method for fermenting chymosin by using nisin fermentation wastewater as a partial nitrogen source, relating to the method for producing the chymosin by microbial fermentation. The invention solves the problems that at current, the method for producing the chymosin by the microbial fermentation has high cost, and the direct discharge of the fermentation wastewater of the nisin results in environment pollution. The method comprises the following steps: 1, preparing a culture medium; 2, preparing rhizomucor miehei seed liquor; 3, inoculating the rhizomucor miehei seed liquor to a triangular flask; and 4, culturing the inoculated seed liquor for 96h at the temperature of 37-38 DEG C to obtain the chymosin. The invention can utilize waste, reduce the environment pollution caused by directly discharging the fermentation wastewater of the nisin, and lower the cost for fermenting the chymosin. The invention can be applied to the field of microbial fermentation.

The invention discloses a rhizomucor miehei strain and an application thereof to preparation of Beta-glucanase and chymosin. The rhizomucor miehei CAU432 disclosed by the invention has a preservation number of CGMCC (China General Microbiological Culture Collection Center) No.4967. The rhizomucor miehei CAU432 disclosed by the invention is good in heat resistance and strain stability, high in enzyme yield, low in fermentation cost (a used carbon source and a used nitrogen source are easy to buy and low in price), short in production peroid, suitable for being applied to the field of industrial production and better in industrial application prospect.

The invention relates to a method of catalytically synthesizing sucrose-6-acetate by Rhizomucor miehei lipase. The method is characterized by comprising the steps of a. adding DMF and tert-pentyl alcohol into a reaction kettle according to a volume ratio of 1:(3 to 5), and performing uniform mixing; b, adding sucrose into a reaction kettle; adding vinyl acetate and the Rhizomucor miehei lipase, controlling the concentration of the sucrose to be 0.03 to 0.12 mmoL / mL, the concentration of the Rhizomucor miehei lipase to be 6 to 60 mg / mL, the mol ratio of the sucrose to the vinyl acetate to be 1:(5 to 20) and the temperature to be 30 to 35 DEG C to react for 8 to 24 h, performing filtration, and placing filter liquid at a room temperature for 20 to 40 min; c, performing reduced pressure distillation on the filter liquid at the temperature of 50 to 70 DEG C and the pressure of -0.080 to -0.1 MPa, cooling liquid in a condensing groove to the room temperature, adding dichloromethane, addingthe dichloromethane again after turbidity occurs, controlling the ratio of the total quantity of the dichloromethane to the volume of the condensing liquid to be (4 to 7):3, performing still standingand filtration to obtain mother liquid; and d, pumping the mother liquid into a centrifugal machine to obtain faint yellow crystals. The method provided by the invention has the advantages that the reaction conditions are mild; the operation is simple; the generation of isomers is avoided; the catalytic selectivity, the product safety and the conversion rate are high; and the cost is low.

Topical compositions comprising extracts of Rhizomucor miehei that are substantially devoid of acid-protease activity and their use in treating dermatologic conditions, including reducing the appearance of signs of skin aging.

The invention discloses a method for preparing Rhizomucor miehei renninum through liquid fermentation of bran, which comprises the following steps: activating a strain, preparing a spore suspension, preparing a liquid fermentation culture medium, inoculating, performing fermentation culture, and preparing an enzyme liquid. The preparation method disclosed by the invention has the advantages of low raw material cost, simple operating process and short fermentation period, lowers the production cost and is suitable for being popularized in production.

The invention relates to rhizomucor miehei alpha-amylase, and an encoding gene and an application of the rhizomucor miehei alpha-amylase. A protein has alpha-amylase activity; the gene of the proteinis efficiently expressed in pichia pastoris; the maximum enzyme activity after high density fermentation of the pichia pastoris is 29794.2U / mL; the specific enzyme activity after purification is 3502.1U / mg; an optimum reaction pH (potential of hydrogen) is 6.0; the protein keeps stable when the pH is 4.5-8.0; an optimum reaction temperature is 75 DEG C; and the protein keeps the higher enzymic activity below 65 DEG C and has better thermal resistance. The amylase can increase a specific volume of steamed bread by 7.7% and retard aging of the steamed bread when the amylase is applied to the steamed bread; and a maltose content can reach 54.1% when the amylase is applied to maltose production. The amylase has good utilization potentiality in the food industry.

Methods and compositions for reducing and / or inhibiting inflammation by topical application of dermatocosmetic compositions comprising effective amounts of extracts of Coriolus versicolor that have been hydrolyzed by an acid protease, preferably Rhizomucor miehei, and thereafter rendered substantially devoid of acid-protease activity.

The invention discloses a production method and applications of a novel rhizomucor miehei aspartic protease, and particularly relates to a recombinant aspartic protease, a coding gene, and a production method and applications of the recombinant aspartic protease. The recombinant aspartic protease is protein having an amino acid sequence shown as SEQ ID NO:1. The coding gene of the recombinant aspartic protease is a DNA molecule shown as SEQ ID NO:2. An aspartic protease gene that is RmproA of rhizomucor miehei is connected to a pichia pastoris GS115 expression vector pPIC9K, and is converted into pichia pastoris and induced expression is performed to obtain the recombinant aspartic protease. Through high-density fermentation in a fermentation tank having a volume of 5 L, the maximum protease production activity at 156 h of a recombinant strain is 3400 U / mL, and the protein content is 6.42 mg / mL. The recombinant aspartic protease is capable of effectively tendering pork, reducing shearing force, making meat palatable, and effectively hydrolyzing animal and plant protein to prepare low-molecular-weight polypeptides.

The invention discloses rhizomucor miehei-derived beta-1,3-glucanase and application thereof. The rhizomucor miehei-derived beta-1,3-glucanase is thermophilic fungus rhizomucor miehei-derived endo-type beta-1,3-glucanase (RmLam81A) that specifically acts on the beta-1,3 glycosidic bond in curdlan polysaccharide. Pichia pastoris is used for heterologous expression, and after high-density fermentation culture, the final enzyme activity is up to 694U / mL. Food grade beta-1,3-glucan (the curdlan polysaccharide, laminarin, and the like) are used as raw materials for enzymatic hydrolysis to obtain beta-1,3-glucosyl-oligosaccharides with degree of polymerization (DP) of 2-5. The yield of the soluble oligosaccharides is about 75-85%. The beta-1,3-glucosyl-oligosaccharides can be obtained by slag liquid separation, material liquid enrichment and spray-drying of hydrolysate. The beta-1,3-glucanase disclosed in the present invention has excellent enzymatic properties. Compared with that of the chemical or enzymatic methods in the prior art for preparing the beta-1,3-glucosyl-oligosaccharides, the method using the rhizomucor miehei-derived beta-1,3-glucanase mild in reaction conditions, environmentally-friendly, and higher in the yield of the oligosaccharides, and has important application value in enzymatic preparation of the beta-1,3-glucosyl-oligosaccharides.

Methods and compositions for reducing and / or inhibiting inflammation by topical application of dermatocosmetic compositions comprising effective amounts of extracts of Coriolus versicolor that have been hydrolyzed by an acid protease, preferably Rhizomucor miehei, and thereafter rendered substantially devoid of acid-protease activity.

The present invention discloses a soy protein preparation method which comprises the following steps: (1) soy protein in aqueous solution is heated at a pH value of 4.5-5.0 and a temperature of 65-75 DEG C for 15-20 min, and then at a pH value of 5.5-5.8 and a temperature of 78-85 DEG C for 25-45 min; (2) rhizomucor miehei (1.5-2% of the soy protein by mass) and carboxymethylcellulose sodium (3-10% of the soy protein by mass) are added and stirred at a pH value of 5-5.5 for 75-95 min, and then stirred again at a pH value of 5.5-6.5 for 65-75 min; (3) enzyme is inactivated, the solution is cooled and the pH value is adjusted to 5-6.8, and then the whole system undergoes ultrasonic processing at 10-15 DEG C and 400-500rpm for 15-20 min; and (4) the final step is freeze drying. The soy protein obtained from the preparation method has good foaming properties and excellent foam stability.

The invention discloses a preparation method of konjaku mannan oligosaccharide and a special beta-mannase mutant adopted by the same. The beta-mannase mutant provided by the invention is from rhizomucor miehei, has the advantages of high stability, high enzymatic specific activity and the like and has very high application value in the industries of foods, feeds and the like. Engineering bacteriaformed by importing the beta-mannase mutant into pichia pastoris are fermented under a high density in a 5L fermentation tank, and the enzymatic activity of fermented liquid can reach 72,600U / mL (protein content is 9.1mg / mL). The beta-mannase mutant is used for hydrolyzing konjaku flour to prepare the konjaku mannan oligosaccharide; mainly mannan oligosaccharide with a polymerization degree beingbetween 2 and 6 is in hydrolysate; a konjaku hydrolysis rate is 90.2%; reducing sugar yield is 69.9%. The invention provides important bases for preparation of the konjaku mannan oligosaccharide by utilizing the konjaku flour.

The invention discloses a rhizomucor miehei strain and an application thereof to preparation of Beta-glucanase and chymosin. The rhizomucor miehei CAU432 disclosed by the invention has a preservation number of CGMCC (China General Microbiological Culture Collection Center) No.4967. The rhizomucor miehei CAU432 disclosed by the invention is good in heat resistance and strain stability, high in enzyme yield, low in fermentation cost (a used carbon source and a used nitrogen source are easy to buy and low in price), short in production peroid, suitable for being applied to the field of industrial production and better in industrial application prospect.

The present invention relates to a mutated Aspergillus oryzae strain. Specifically, the invention relates to a mutated Aspergillus oryzae strain of which the productivity is improved by being compared with a non-mutated strain, endogenous enzymes, such as amylase and / or foreign protein, and food-purpose lipase, preferably the food-purpose lipase selected from rhizomucor miehei lipase, thermomyces lanuginosus lipase and fusarium oxysporum lipase. The invention also relates to various applications of the strain.

The invention discloses a wood fiber degradation complex microbial inoculant of mixture of livestock and poultry manure and straw. The complex microbial inoculant is prepared from rhizomucor miehei NJAU-F4-5 fermentation liquor with the concentration of 109 / ml and the preservation number of CGMCC No.18135 and brevibacillus reuteri NJAU-N2 with the concentration of 109 CFU / ml and the preservation number of CGMCC No.18020. The wood fiber degradation complex microbial inoculant has the following beneficial effects: the production efficiency of organic fertilizer is improved, the economic benefitsand the ecological environment benefits of enterprises are improved, meanwhile, the straw degradation rate of the complex microbial inoculant reaches 20.55%, the straw degradation effect of the complex microbial inoculant is better than that of single bacteria and bacteria-free bacteria, the degradation time can be shortened, the degradation process is accelerated, and the production cost is saved.

The invention discloses a fully synthetic culture medium and a method for producing rhizomucor miehei lipase by a fermentation method. The molar weight ratio of the carbon element to the nitrogen element in the total synthesis culture medium is 11-18; the total synthesis culture medium comprises a carbon source and a nitrogen source, and the carbon source is cane sugar, trehalose, lactose or fructose; the nitrogen source is alanine. According to the total synthesis culture medium for producing rhizomucor miehei lipase through aspergillus oryzae fermentation, sucrose, trehalose and the like are used as a carbon source, alanine is used as a nitrogen source, the carbon source and the nitrogen source are cooperatively added and used according to a scientific carbon-nitrogen ratio, the carbon source and the nitrogen source have an obvious interaction effect, and the aspergillus oryzae can be effectively promoted to synthesize the rhizomucor miehei lipase. The activity of the rhizomucor miehei lipase obtained through fermentation culture of the fully-synthetic culture medium reaches up to 60.42 U / ml, and the effect is remarkable.

The invention provides a chiral resolution method of an isobutyrate compound. The isobutyrate compound is a racemic isobutyrate compound and has the following structural general formula, R1 is selected from any one of the following substituted or unsubstituted groups: pyridine, pyrazole, pyrimidinone, 4hydroxyquinoline, quinazoline and quinazolinone, and R2 is selected from substituted or unsubstituted alkyl and aryl. According to the method, a racemic isobutyrate compound is subjected to hydrolysis treatment by adopting hydrolase to obtain a hydrolyzed system, and the hydrolase is from Thermomyces sp., Rhizomucor miehei and Bacillus sp.; and step S2, separating acid in the hydrolyzed system from the unhydrolyzed isobutyrate compound. The hydrolase used in the method is good in stability,good in enantiomer selectivity, mild in reaction condition, low in cost and simple to operate.

This patent introduces a chiral resolution method for obtaining (RR)- and (SR)-loxoprofen enantiomers, that is, an enzymatic stereoselective esterification method for splitting loxoprofen partial enantiomers The invention belongs to the technical field of splitting chiral compounds by biological methods. Choose Novozym from Rhizomucor miehei ® 40086 lipase is used as a biocatalyst, and the lipase has the characteristics of high selectivity and high catalytic efficiency in the catalytic esterification of loxoprofen, and can obtain (RR)-loxoprofen and (SR) with high optical purity ‑Loxoprofen. After enzyme-catalyzed stereoselective esterification resolution, the enantiomeric excess value of the target product reached 98.64%, the yield was 72.70%, and the overall conversion rate of the substrate was 63.40%. Compared with other methods, the method has high reaction concentration, fast reaction rate, high optical purity of the product, simple operation and post-treatment, low cost, and green and environment-friendly production process, and is an efficient resolution method for loxoprofen.

The invention relates to a sulfur-containing flavor ester composition and a preparation method and cheese essence thereof. The preparation method comprises the following steps: (1) taking anhydrous milk fat and 3-methylthio-propanol as raw materials, and adding 200-400U of rhizomucor miehei lipase into the raw materials per gram for performing enzymatic hydrolysis, thereby obtaining an enzymolysis product; (2) performing level-I molecular distillation on the enzymolysis product obtained in the step (1), wherein the vacuum degree of level-I molecular distillation is 30-40Pa, removing the fraction on a temperature section of 50 DEG C or below, and continuously performing level-II molecular distillation, wherein the vacuum degree of level-II molecular distillation is less than 5Pa, collecting a light phase part of the fraction on a temperature section of 50-180 DEG C, thereby obtaining the product. According to the invention, the preparation method is simple, and the prepared sulfur-containing flavor ester composition has aroma of cheese and cream cheese after high-temperature processing. The cheese essence prepared by taking the sulfur-containing flavor ester composition disclosed by the invention as a raw material is strong in flavor, mellow, natural and good in high-temperature stability.

The invention provides a chiral resolution method of an isobutyrate compound. The isobutyrate compound is a racemic isobutyrate compound, and has the following structural general formula: R1 is selected from any one of the following substituted or unsubstituted groups: pyridine, pyrazole, pyrimidone, 4-hydroxyquinoline, quinazoline and quinazolinone, R2 is selected from substituted or unsubstituted alkyl and aryl, and the chiral resolution method comprises the following steps: S1, carrying out hydrolysis treatment on a racemic isobutyrate compound by adopting hydrolase to obtain a hydrolyzed system, wherein the hydrolase is derived from Thermomyces sp.Rhizomucor miehei, Bacillus sp.; and S2, separating the acid in the hydrolyzed system from the isobutyrate compound which is not hydrolyzed. The hydrolase used in the method has the advantages of good stability, good enantiomer selectivity, mild reaction conditions, low cost and simple operation.

The present invention provides an efficient wood fiber degradation composite bacteria agent and an application thereof in composting. For further improving the composting effect of livestock and poultry breeding feces and straw wastes, a composite bacteria agent containing Rhizomucor miehei NJAU-F4-5, Brevibacillus reuszeri NJAU-N20 and bacillus sp. NJAU-N30 is developed, thereby providing composite bacteria resources and technical support for efficient composting of the livestock and poultry breeding feces and the straw wastes. The composite bacteria agent is applied to acceleration of composting fermentation. A fermentation process comprises the following steps of uniformly mixing the livestock and poultry breeding feces and straws according to the certain C / N, inoculating the liquid composite bacteria agent, building a strip-shaped stack, and by adopting a strip-shaped stack type fermentation process, performing turning-throwing fermentation. Compared with the non-addition of bacteria and the addition of single bacteria for treatment in a control, the addition of the composite bacteria agent effectively promotes the temperature rise of a compost body in the initial stage of composting, facilitates the compost body to enter the cooling stage in advance, promotes the increase in germination index of a compost product and effectively drives the compost maturity.

The invention discloses a method for efficiently producing rhizopus oryzae lipase by using aspergillus oryzae. The method comprises the following steps of: seed culture: inoculating a spore suspension of the aspergillus oryzae into a seed culture medium, and culturing under the conditions that the temperature is 26-30 DEG C and the rotating speed is 100-200rpm to obtain a seed solution; fermentation culture: inoculating the seed solution into a fermentation culture medium, fermenting under the conditions that the temperature is 26-30 DEG C and the initial rotating speed is 300rpm, and obtaining fermentation liquor after the fermentation is finished, namely obtaining the rhizopus oryzae lipase, wherein the initial nitrogen source amount in the fermentation culture medium is 10-15% of all nitrogen sources, and the nitrogen sources are supplemented in the subsequent fermentation process. By adopting the method disclosed by the invention, the bacterial form can be controlled, a stable fermentation system is obtained under the conditions of nitrogen source supplementation and rotating speed control, the effective volume of fermentation is obviously improved, and the RML yield and enzyme activity are improved.