44results about How to "Improve enzyme production efficiency" patented technology

The invention relates to a construction method for co-expression hemoglobin VHb and cellulase protein in pichia pastoris, in particular to cellulase protein gene codon optimization and construction of a cellulase protein and vitreoscilla hemoglobin protein (VHb)-co-expressed pichia pastoris system. According to the construction method, codon bias optimization is performed on the nucleotide sequence of EG II (GenBank Accession No.DQ178347.1) through Gene Designer (DNA2.0, Menlo Park, CA, USA) software, a pPIC9K-eg2 expression vector is constructed, and a recombined pichia pastoris strain is obtained by taking Pichiapastoris GS115 as a host through electrotransformation. In addition, the nucleotide sequence of the VHb (GenBank Accession No.M30794.1) is obtained from NCBI and artificially synthesized into a gene, then a pPICZalphaA-vgb expression vector is constructed, and the co-expressed pichia pastoris strain is obtained by taking the recombined pichia pastoris strain containing the EG II gene as a host through electrotransformation. Detection shows that the co-expressed strain is improved on the aspects of bacterial concentration growth and enzyme activity, wherein the OD600 value is increased by 7.2%, and the enzyme activity is improved by 2.2%.

The invention discloses a bacillus licheniformis and a method for utilizing the sludge as raw material for producing alkaline protease. The bacillus licheniformis is gram-positive bacteria and is shaped like a straight rod, and the collection number is CGMCC 1970. The method for utilizing the bacillus licheniformis for producing the alkaline protease includes (1) the conditioning of a culture medium; (2) shaking culture; (3) the fermentation by a fermentation tank; and (4) the purification of the protease. The bacterial strain of the invention is utilized for producing the alkaline protease, which can not only dispose sludge, but also obtain the alkaline protease product with high additional value and lower cost. The method uses urban waste sludge to replace the traditional food culture medium for producing the alkaline protease product, which not only provides a new way for the resource disposal of urban sludge, but also can greatly reduce the price of the product and promote the industrial production and application of protease preparations.

The invention belongs to the technical field of enzyme engineering and particularly relates to a method for obtaining sucrose isomerase through site-directed mutation so as to improve the product specificity of the sucrose isomerase, and a method for preparing immobilized sucrose isomerase and achieving separation of the immobilized enzyme through a non-aqueous phase catalytic system. The content of isomaltulose in a catalytic product of the sucrose isomerase is substantially increased and increased to 99.16% from 90.28%, and the enzyme preparation process is easy to implement and is high in enzyme producing efficiency; due to the sucrose isomerase immobilization technology, the sucrose isomerase can easily keep high catalytic activity and low enzyme amount loss in the repeated catalysis process, multi-batch continuous use can be performed, operating difficulty can be greatly reduced, and economic cost is saved.

The invention discloses a high-temperature-resistant high-yield cellulase bacillus subtilis and application thereof. A cellulase production strain is separated from surface soil of a corn straw pile in a Jiangjin area of Chongqing, and the preservation number of the bacillus subtilis is CCTCC NO: M 2020002; a strain Z2 with the best enzyme production performance is finally obtained through enrichment culture, a CMC-Na plate experiment, a lignin plate experiment and an enzyme production experiment, and the obtained strain is subjected to 16S rRNA sequence comparison and identification to determine that the strain is bacillus subtilis Z2. By optimizing the culture medium and the culture conditions, the filter paper enzyme activity, the CMC enzyme activity and the beta-glucosidase activity under the optimal enzyme production conditions of 0.800 U / ml, 5.20 U / ml and 2.07 U / ml are obtained respectively. Cellulase produced by the strain has relatively high activity and stability under the conditions of a buffer solution with the pH value ranging from 4.0 to 10.0 and the temperature of 30 DEG C and 80 DEG C. Cellulase produced by the strain is mixed with commercial xylanase, then a saccharification experiment is carried out on pretreated straw stalks, and 84.27 mg / ml of total reducing sugar can be obtained.

The invention discloses a method for increasing the output of alkaline pectinase produced by fermentation by adding sorbitol, relating to the technical field of application of the carbon source adding policy in the optimization of fermentation. In the method, recombinant Pichia pastoris GS115 is adopted as fermentation strains, and sorbitol is added into the fermentation liquid at a constant speed or accelerated speed during fermentation induction phase to improve the activity of cells, weaken the toxicity of methanol, and increase the output of the alkaline pectinase. The lower adding amount of sorbitol can not inhibit the enzyme production effect of methanol to the strains, on the other hand, the toxicity to the strains from the methanol can be reduced to a certain extent, and the efficiency of enzyme production is further increased. When the sorbitol is added at a varying speed (0.9g / h-7.2g / h), the activity of enzyme is increased from 930U / ml to 1541U / ml. The invention is simple and effective in process, capable of being applied in the industrial production of pectinase in the future, and instructive to the optimization of fermentation of other Pichia pastoris.

The invention relates to an inulase mutant and a preparing method. According to the inulase mutant and the preparing method, directed evolution is carried out on an encoding gene inuI of inulase in vitro with a codon optimization technology, an error-prone PCR technology and a site-directed mutagenesis technology, and the mutant with the inulase activity remarkably improved is screened out. The K<cat> value of the mutant is 3.1 times that of the inulase of an original strain; in addition, the preparing process of the inulase is easy to implement and high in enzyme production efficiency, and the enzyme activity of fermentation liquor in a 30-m<3> system can be 60,000 U / ml. Meanwhile, the invention discloses a DNA sequence, an expression vector and a host cell of the encoding inulase mutant. According to the inulase mutant, the preparing method and the DNA sequence, the expression vector and the host cell of the encoding inulase mutant, high efficiency and simple and convenient operability are achieved.

The invention discloses a fungus-derived laccase gene, and relates to the technical field of biology. The laccase gene is lac1, and the cDNA sequence of the lac1 is as shown in SEQ ID NO.2, or the lac1 has a base sequence for encoding an amino acid sequence as shown in SEQ ID NO.3. The invention also provides a recombinant pichia pastoris engineering strain carrying a laccase gene expression vector and an application of the recombinant pichia pastoris engineering strain. The fungus-derived laccase and the recombinant pichia pastoris engineering bacteria thereof have the beneficial effect that the laccase Lac1 newly identified in Trametes sp. Is expressed in a heterologous manner. The obtained recombinant laccase has good thermal stability and good tolerance to metal ions, organic solvents and inhibitors, can catalyze decoloration of indigo blue and other dyes, and has a good application prospect in industry.

The invention discloses a bacillus licheniformis and a method for utilizing the sludge as raw material for producing alkaline protease. The bacillus licheniformis is gram-positive bacteria and is shaped like a straight rod, and the collection number is CGMCC 1970. The method for utilizing the bacillus licheniformis for producing the alkaline protease includes (1) the conditioning of a culture medium; (2) shaking culture; (3) the fermentation by a fermentation tank; and (4) the purification of the protease. The bacterial strain of the invention is utilized for producing the alkaline protease, which can not only dispose sludge, but also obtain the alkaline protease product with high additional value and lower cost. The method uses urban waste sludge to replace the traditional food culture medium for producing the alkaline protease product, which not only provides a new way for the resource disposal of urban sludge, but also can greatly reduce the price of the product and promote the industrial production and application of protease preparations.

The invention provides an excrement treatment composition, a preparation method and application thereof, and a method for treating anhydrous ecological toilet excrement by using the excrement treatment composition. The excrement treatment composition comprises a complex microbial inoculant and a biological padding, the complex microbial inoculant comprises, by mass, 1-3 parts of aspergillus, 3-7 parts of bacillus subtilis, 5-9 parts of saccharomycetes and 1-4 parts of trichoderma, and the biological padding comprises crop straw. By combining the characteristics of the biological padding and the excrement, the preparation of the complex microbial inoculant is optimized, organic macromolecular substances such as protein in the excrement can be efficiently decomposed, the fermentation periodis shortened, and the frequency of cleaning toilets is reduced. When the excrement treatment composition is used for excrement treatment, ammonia gas and other peculiar smells can be effectively removed, and the excrement treatment composition is suitable for indoor use and has important significance in application and popularization of waterless ecological toilets.

The present invention relates to a composition that can catalyze the degradation of uric acid in the intestinal tract. The composition contains Cordyceps militaris fungus powder or dry enzyme powder, and the Cordyceps militaris fungus powder or dry enzyme powder contains uric acid oxidase that can catalyze the degradation of uric acid. . The present invention also provides the preparation method and application of the composition of the present invention that can catalyze the degradation of uric acid in the intestinal tract. The composition of the present invention can catalyze the degradation of uric acid into allantoin, reduce the uric acid content in the intestinal tract, increase the ratio of uric acid secretion and excretion in the intestinal tract, thereby reducing the concentration of uric acid in the blood, prevent and treat hyperuricemia and Gout disease; at the same time, it also reduces the excretion of uric acid in urine and reduces the incidence of uric acid stones.

The invention discloses a Sartorya Vuill W-10 with high cellulase production, with a preservation No. CGMCC No. 11991. The invention can obtain high-yield cellulose crude enzyme liquor by fermentationof Sartorya Vuill W 10, wherein the filter paper enzyme activity of the cellulase can reach 1.26 U / ml; The cellulase from fermentation can be used in the production of bioethanol, which can effectively improve the enzymatic hydrolysis rate of straw residue and ethanol yield, and has a certain application value. The invention not only has low production cost and high ethanol yield, which is favorable for commercial application and implementation of bioethanol, but also provides a way for comprehensive utilization of waste organisms.

The invention relates to the technical field of microorganisms, in particular to a bacillus velei and its application. The preservation number of Bacillus Velez Lpl-wx in the present invention is CGMCC No.17045. The bacterial strain can be applied to the preparation of astaxanthin esterase, and because the Bacillus velaisi belongs to prokaryote, the fermentation process is fast and the enzyme production efficiency is high, and the astaxanthin esterase can be obtained quickly and efficiently. The strain can be further applied to the preparation of astaxanthin monomers, which can effectively degrade astaxanthin esters and obtain astaxanthin monomers, providing a green and efficient catalytic pathway with less by-products, which is a good catalyst for the production of astaxanthin monomers. Preparation and promotion provide the technical basis.

The invention discloses a method for increasing the output of alkaline pectinase produced by fermentation by adding sorbitol, relating to the technical field of application of the carbon source adding policy in the optimization of fermentation. In the method, recombinant Pichia pastoris GS115 is adopted as fermentation strains, and sorbitol is added into the fermentation liquid at a constant speed or accelerated speed during fermentation induction phase to improve the activity of cells, weaken the toxicity of methanol, and increase the output of the alkaline pectinase. The lower adding amountof sorbitol can not inhibit the enzyme production effect of methanol to the strains, on the other hand, the toxicity to the strains from the methanol can be reduced to a certain extent, and the efficiency of enzyme production is further increased. When the sorbitol is added at a varying speed (0.9g / h-7.2g / h), the activity of enzyme is increased from 930U / ml to 1541U / ml. The invention is simple and effective in process, capable of being applied in the industrial production of pectinase in the future, and instructive to the optimization of fermentation of other Pichia pastoris.