119 results about "Industrial enzymes" patented technology

The invention provides a nicotinamide ribokinase mutant and application thereof. Compared with an amino acid sequence SEQID NO. 2, the difference of the amino acid sequence of the mutant is that the D45th, D58th, R161th and Y164 sites in the amino acid sequence SEQID NO. 2 are subjected to single mutation or in-pair combined mutation or three combined mutation or four combined mutation. Novel nicotinamide ribokinase mutant industrial enzyme is used for synthesis and preparation of beta-nicotinamide mononucleotide. The constructed nicotinamide ribokinase mutant enzyme has the advantages that the enzyme cost is low, the transformation time is short, and the process operation is simple, and has a broad large-scale industrial application prospect.

The invention relates to a host cell and a method for the host cell applied in efficient secretion expression of a recombinant protein, which belongs to the filed of microbiological engineering and fermentation engineering. The invention provides a bacillus licheniformis host cell CBB3008 which is preserved in the China Center for Type Culture Collection with the preserving number of CCTCC NO: M 208236. The invention transforms an expression plasmid of an industrial enzyme which is obtained by a gene cloning technology into the host cell to synthesize an industrial enzyme preparation in the host cell efficiently, and then secretes the synthesized enzyme protein into a culture medium efficiently through a protein secretion system of the host cell, thus the invention can guide efficient secretion production of the industrial enzyme preparation. Therefore, the invention is helpful to reduce the fermentative production cost of the industrial enzyme preparation, simplify the fermentative production process and reduce the environmental pressure of the fermentation industry. A method for host cell screening and genetic improvement of the invention can also be used for other types of host cells, in particular for the breeding of the host cells of bacillus subtilis, bacillus megaterium, bacillus pumilus, bacillus starch solution and the like.

The invention discloses a method for producing a wastewater denitrification carbon source through lactic acid fermentation of kitchen waste, and belongs to the technical field of solid waste resourceful treatment. The method comprises the steps of adopting kitchen waste as a raw material; sorting and impurity removal, controlling the solid-to-liquid ratio, grinding, pulping, and carrying out quality-based and phase-splitting treatment; after the waste oil is heated and recycled, the residual garbage slurry is injected into an anaerobic fermentation tank for batch lactic acid fermentation, after fermentation is finished, solid-liquid separation is carried out, lactic acid-containing supernatant can be directly used as a liquid carbon source in the sewage denitrification stage, a good biological denitrification effect is achieved, nitrogen-containing residues can be collected and recycled, and full recycling is achieved. According to the invention, kitchen waste is directly used for efficient open lactic acid fermentation; according to the method, the indigenous microbial flora in the kitchen waste is fully utilized to hydrolyze the substrate and efficiently and directionally convertthe substrate into lactic acid, exogenous nutrition and industrial enzyme preparations do not need to be added, the process is simple, the production cost is low, a cheap carbon source is provided for a denitrification section of a sewage treatment plant while resource utilization of the kitchen waste is realized, and the biological denitrification efficiency is improved.

The invention discloses bacillus tequilensis CGX5-1 which is preserved with the number CGMCC No.5935 in China General Microbiological Culture Collection Center on March 26, 2012. The bacillus tequilensis has high capability of producing beta-1, 3-1, 4-glucanase, and the bacillus tequilensis and good genes thereof are applied to the field of production of industrial enzyme preparations and have good prospects in reduction of cost for producing the beta-1, 3-1, 4-glucanase and improvement on enzyme preparation quality.

The present invention relates to the field of microbial engineering technology. The enzyme preparation is prepared through the process including slant culture and storing, shaking culture, seeding tank culture and fermenting tank production of high enzyme activity bacteria strain. Bacillus subtilis of No.13 strain and preservation number CCTCC No.M200038 is selected. The hemp degumming process includes the fermentation of enzyme liquid filtering residue, diluting bacteria liquid to 3-6 times, regulating the pH value of fermented enzyme liquid in degluing tank to 7.5-9.0, hemp material pre-treatment, degumming with degumming liquid for 2-10 hr. The process of the present invention has no environmental pollution, and can produce hemp top with length, fineness and breaking strength superior to those of chemical process.

The invention discloses gene engineering bacteria with a high yield of sucrose isomerase, and a method for efficiently producing the sucrose isomerase, and belongs to the technical field of gene engineering and fermentation engineering. According to the method, codon and signal peptide optimization is adopted to ensure that the expression quantity of a sucrose isomerase gene in bacillus licheniformis is improved by more than or equal to 3 times compared with the original expression quantity. Finally, a growth medium and a fermentation process suitable for synthesis and secretion of an enzyme, and recovery and preparation of a product are established, so that the content of isomaltulose in the product reaches 100%. Therefore, a firm foundation is laid for the application of the sucrose isomerase in food and other industries, and a technical support is provided for efficient preparation of other industrial enzyme preparations.

The invention discloses genes and engineering bacteria of acid and high temperature resistant alpha-amylase and a preparation method of the genes and engineering bacteria. Compared with a mutant reported by literature, under the condition that the pH ranges from 4.2 to 4.6, and the temperature is 95 DEG C, the thermostability of the acid and high temperature resistant alpha-amylase is enhanced, the expression quantity is 50 times that of existing acid and high temperature resistant alpha-amylase, the alpha-amylase is applied to the fields of chemical industry, food, medicine and the like, starch can be efficiently degraded under the conditions of strong acidity and high temperature, the process can be simplified, environmental pollution is reduced, and a wide application prospect is achieved. Meanwhile, according to genes and engineering bacteria of the acid and high temperature resistant alpha-amylase and the preparation method of the genes and engineering bacteria, rational design of applied combined construction prediction has important guiding significance in improving thermostability of an industrial enzyme.

A high temperature alpha-amylase and a high efficiency preparation method of the mutant thereof belongs to the fermentation engineering field. The invention provides a method for high efficiently secreting expressing a high temperature alpha-amylase and mutant thereof with alpha-amylase activity through a bacillus, especially a bacillus licheniformis mutant strain as a host cell. The synthesis and secretion level of the high temperature alpha-amylase or mutant thereof of the recombinant strain of the invention reaches 18-25mg/mL during fed-batch fermentation and is 3-4.5 times of the original strain high temperature alpha-amylase synthesis level. The high temperature alpha-amylase of the invention is useful for reducing the fermentation manufacture cost of the industrial enzyme, simplifying the fermentation manufacture process and reducing fermentation industrial environment pressure.

The invention relates to an optimization method of culture conditions of alkaline pectinase gene engineering bacteria, comprising the following steps: (1) adopting a single-factor experiment to optimize the fermentation medium formation of the alkaline pectinase gene engineering bacteria to obtain a preliminarily optimized fermentation medium; (2) optimizing a target strain medium with a response surface methodology to obtain the target strain medium; and (3) optimizing the fermentation culture conditions to the target strain by a post-optimizing medium to obtain the optimization culture conditions. The invention gropes a set of optimal fermentation culture conditions to ensure that enzymatic activity produced by engineering bacteria fermentation reaches 758.7825U/ml which is improved by 50 times compared with the activity of alkaline pectinase produced by original wide strains and is improved by 3 times compared with the enzymatic activity of alkaline pectinase produced by LB fermentation medium fermentation gene engineering bacteria, so that the invention has an important meaning for improving the market competitiveness of industrial enzyme.

The invention discloses a keratinase mutant with improved catalytic performance and an application, belongs to the technical field of genetic engineering and provides recombinant keratinase constructed on the basis of leading peptide engineering and multi-site saturated mutation and a leading peptide engineering method. A recombination strain M7 shows high keratinase catalytic potential, enzyme activity is remarkably improved from 179 U/mL to 1114 U/mL, the optimal reaction temperature is 40 DEG C, the optimal reaction pH is 10.0, over 80% enzyme activity is retained after water bath processing is performed for 90 min at 40 DEG C, heat stability is better, and the keratinase mutant is suitable for industrial large-scale production. Besides, the method for modifying protease with the leading peptide engineering can be a universal way for modifying industrial enzyme with the self-shearing characteristic.

The invention provides a method for preparing lactose enzyme which is a novel high-efficient preparation. In the method, bacteria are processed by NTG with a certain concentration to produce high yield bacteria, solid fermentation is carried out under the condition which is suitable for the growth of microorganisms of the high yield bacteria, and industrial enzyme with high purity quotient and ideal yield coefficient is obtained by extracting and separating under a totally enclosed condition. The method has the advantages of soft preparing condition and little generation of three wastes.

The invention relates to a method for preparing clove oil microcapsules employing cyclodextrin mother solution, belonging to the technical field of the preparation of clove oil microcapsules. The method comprises the following steps: using the waste cyclodextrin mother solution in the industrial enzyme conversion preparation method of beta-cyclodextrin as raw material, treating with activated carbon, selecting proper inclusion process and adopting effective ingredients such as beta-cyclodextrin and the like in the mother solution to pack clove oil which is volatile flavoring fragrancy industrial material and preparing clove oil microcapsules. Compared with the clove oil microcapsules prepared from the finished beta-cyclodextrin, the product of the invention has the similar shape and properties. The cost of the method of the invention is much lower that that of the traditional method so as to realize the effective reuse of waste and the method also reduces the pollution to the environment.

The invention discloses an enzyme-metal ion nano compound and a preparation method thereof.The preparation method includes following steps: drying a water solution of metal ions and enzyme to obtain the enzyme-metal ion nano compound, wherein the metal ions are one of calcium ions, magnesium ions, iron ions, ferrous ions, copper ions, barium ions, manganous ions, bivalent nickel ions and zinc ions.An enzyme immobilization method has the advantages of simple operation and low cost.By controlling concentration of the metal ions and the enzyme in the solution, the enzyme-metal ion nano compound prepared by the method is not generated in a precipitate form while a nano compound of 500-1200 nm soluble in the water solution is formed, stability of the enzyme in the nano compound is improved remarkably, and enzyme activity is basically maintained unchanged and even is improved.Due to above characteristics, the enzyme-metal ion nano compound is different from conventional nano carrier immobilized enzyme, and has good application prospect in the field of industrial enzyme catalysis, biosensors and biomedicine.

The invention discloses a method for preparing an oligopeptide refreshing beverage by using endogenous endopeptidase and exopeptidase to hydrolyze sesame protein, and belongs to the technical field ofplant protein processing. The method comprises the following steps: sesame seeds are soaked at low temperature and peeled, pulping is performed, filtration is performed to obtain water extracts of peeled sesame seeds, centrifugation is performed on the water extracts to obtain a light phase, an intermediate phase and a heavy phase, protein in the intermediate phase is hydrolyzed by using the sesame endogenous endopeptidase and exopeptidase, centrifugation is performed to remove oil in the intermediate phase, the obtained supernatant and other materials are mixed, sterilization is performed, and packaging is performed to obtain the sesame oligopeptide refreshing beverage. The sesame oligopeptide refreshing beverage prepared by the method has the fragrant smell of the sesame seeds, and notonly is easier to digest and absorb, but also contains some physiologically active peptides; and the method provided by the invention does not use any industrial enzyme preparations or organic solvents, and the preparation process is environmentally friendly.

The invention discloses an avirulent Bacillus cereus strain pBC-1, which is preserved in China General Microbiological Culture Collection Center (CGMCC, in Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard No.1, Beicheng West Road, Zhaoyang District, Beijing City, China, 100101), with an accession number of CGMCC No. 9535. Identification results show that the sequence of the strain pBC-1 is Bacillus cereus. Results of analysis at the levels of genes, protein and cells, the strain pBC-1 provided by the invention does not generate any toxin selected from a group consisting of cereulide, hemolysin BL (Hbl), non-hemolytic enterotoxin (Nhe) and cytotoxin K (CytK). The strain has wide application potential and can be applied to medicine and health care, feed additives or biological veterinary drugs as a probiotic candidate bacterial strain and used for removing pollutants in water bodies and preparing and purifying industrial enzymes.

The invention discloses a solution blending device for industrial enzyme preparation. The device structurally comprises a connecting pipe, a stirring tank, a transmission pipe and supports, the connecting pipe is horizontally connected to the left and right sides of the upper end of the stirring tank, the transmission pipe is located at the front end of the stirring tank, and the supports are fixedly embedded and connected to four corners of the lower end of the stirring tank; the stirring tank is composed of a stirring tank, spraying devices, a rotating shaft and transverse rods, the spraying devices are fixedly connected to the left end and the right end of the inner side of the stirring tank in an embedded mode, the rotating shaft is located between the spraying devices, and the transverse rods are movably clamped to the left side and the right side of the rotating shaft. When new raw materials are mixed and sprayed, an overturning mechanism is extruded to swing outwards to a vertical state, while swinging, a sliding block can drive a magnetic block to slide along with the inclined gradient under the limitation of a sliding rod, when the magnetic block slides, a removing block can be dragged to move towards a collecting device from the upper end of the overturning mechanism, and a pushing plate makes contact with the surface of the overturning mechanism in the moving process to remove mixed liquid adhering to the surface, and the mixed liquid is avoided from mixing with new raw materials.

The invention relates to the technical field of preparation of an industrial enzyme preparation, and particularly relates to a mixing device for preparing an industrial enzyme preparation. The mixingdevice comprises a mixing box, wherein supporting legs are fixedly mounted on the lower end face of the mixing box, a box cover is in threaded connection with the upper end of the mixing box, a main stirring piece is fixedly mounted on the upper end face of the box cover, a containing box is fixedly installed on one side of the upper end face of the box cover, the main stirring piece comprises a limiting sleeve, an installation plate, a motor, a first gear, a second gear, a connecting shaft, a sleeving ball, a driving shaft, stirring blades, a third belt, a linkage disc and a driving plate, the limiting sleeve is fixedly installed in the middle of the box cover, and the installation plate is fixedly installed on the upper end face of the box cover. The mixing device is good in mixing effect and worthy of popularization.

The invention discloses an enzymatic thermal stability improving method. Fusion expression of an amphiphilic short peptide and recombinase is carried out to obtain thermal stability-improved recombinase. The optimal sequence of the amphiphilic short peptide is DWLKAFYDKVAEKLKEAFKVQPYLDDWLKAFYDKVAEKLKEAF. The method has the advantages of substantial effect, simple process and convenient popularization, and provides a new method and idea for the rapid improvement of the thermal stability of industrial enzymes.

The invention relates to the field of eukaryotic expression of industrial enzymes, and particularly relates to a method of producing glycosylated 5-hydroxymethyl furfural oxidase (HMFO) by means of pichia pastoris. According to the codon preference of the pichia pastoris, the gene sequence of the HMFO is optimized; the optimized sequence is connected to a pPICZa-A carrier to convert pichia pastoris X-33. Pichia pastoris having high-copy HMFO gene is then screened, thereby achieving high-effective secretion expression of the HMFO in the pichia pastoris. In addition, the purity of the expressed protein is high and purification steps are simple. Meanwhile, the protein, when being subjected to galactosylated modification, is higher in activity than other proteins in the prior art. The preparation process is low in energy consumption and is high in efficiency and has simple and green production process. The glycosylated HMFO has excellent industrial application prospect.