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138 results about "Isomerase Gene" patented technology

Isomerase Genes encode enzymes (Isomerases) that catalyze spatial or structural changes within a molecule by rearrangement or transfer of specific atoms or moieties to a new intramolecular location to form a new single product. The reactions do not involve a net change in the concentrations of compounds other than the substrate and the product. (NCI)

Genetically engineered bacterium for producing N-acetylneuraminic acid as well as construction and application of genetically engineered bacterium

The invention belongs to the technical field of genetic engineering, and particularly relates to a genetically engineered bacterium for producing N-acetylneuraminic acid through xylose induction. Escherichia coli is used as a starting strain, an N-acetylglucosamine synthesis pathway is integrated on a genome, an N-acetylglucosamine 2-epimerase gene bAGE and an N-acetylneuraminic acid synthetase gene neuB from collar algae are introduced, an N-acetylneuraminic acid synthesis pathway is constructed, and a key gene nano ATEK of a catabolism pathway of the N-acetylneuraminic acid is knocked out. Meanwhile, metabolic pathways of precursor substances required by synthesis of the N-acetylneuraminic acid are subjected to multi-copy reinforcement, part of bypass metabolic pathways are knocked out, key enzyme genes for producing GlcNAc and Neu5Ac are optimized according to different copy numbers, the optimal proportion of the key enzyme genes is finally determined, and the high-yield strain of the N-acetylneuraminic acid is obtained. The highest yield of the N-acetylneuraminic acid can reach 28g / L, the highest production intensity can reach 0.67 g / (L*h) which is the highest value reported at present, and the N-acetylneuraminic acid has important industrial application value.
Owner:TIANJIN UNIV OF SCI & TECH

Immobilization method for cells containing glucose isomerase

The invention discloses an immobilization method for cells containing glucose isomerase. The immobilization method comprises the following steps of adding wet bacteria obtained by performing fermented culture on a recombinant genetic engineering strain containing glucose isomerase genes in buffer liquor so as to obtain bacterial suspension; adding a carrier in the bacterial suspension; uniformly stirring the bacterial suspension; adding polyethyleneimine and glutaraldehyde to perform crosslinking; filtering the bacterial suspension after performing stirring crosslinking on the bacterial suspension for 1-2 hours at the temperature of 0-30 DEG C; washing filter cakes by using distilled water; extruding the filter cakes into long strips by using an axial extruder; airing the long strips at room temperature; and smashing the long strips into granules so as to obtain immobilized glucose isomerase cells containing the glucose isomerase. The immobilization method has the advantages that the cost of an immobilization material is low, the method is easy to operate and high in mechanical strength, the immobilization material is stable at high temperature, prepared recombinant escherichia coli whole cells for producing the glucose isomerase in an immobilization manner can be used for catalyzing D-glucose to produce D-fructose at high temperature, the conversion rate is 54% at the temperature of 85 DEG C, after being repeatedly used ten times, the cells still have enzyme activity which is higher than 90%, and the industrial application prospect is high.
Owner:ZHEJIANG UNIV OF TECH

Engineered Escherichia coli and method of synthesis of catalyzing fumaric acid from maleic acid in presence of Engineered Escherichia coli

The invention relates to engineered Escherichia coli capable of high-yield production of fumaric acid; specifically, fumarase coded genes fumA and fumC of Escherichia coli are knocked off, and maleic cis-trans isomerase gene from Serratia marcescens is converted to obtain the engineered Escherichia coli. The invention also discloses a method of synthesis of catalyzing fumaric acid from maleic acid by using the engineered Escherichia coli. The method includes culturing the engineered Escherichia coli by fermenting, subjecting the engineered Escherichia coli to high-density inductive expression when OD600 reaches 40-80, and applying the obtained fermentation broth to biochemical catalysis of maleic acid in presence of Engineered Escherichia coli to obtain fumaric acid. The genes fumA and fumC in the Escherichia coli are knocked off by using Red homologous recombination, a metabolic pathway of its fumaric acid to L-malic acid is cut off, the maleic cis-trans isomerase from Serratia marcescens is then expressed, and the obtained genetically-engineered Escherichia coli can efficiently convert maleic matrix to synthesize high-purity fumaric acid, there is nearly no L-malic acid byproduct synthesized, and basis is provided for the industrialized production of fumaric acid by whole-cell process catalysis of maleic acid.
Owner:JIANGNAN UNIV

Glucose isomerase and gene, mutant, engineering bacteria and application thereof

InactiveCN104745563AIncrease productionSolve the problem of low optimum reaction temperatureBacteriaMicroorganism based processesBiotechnologyEscherichia coli
The invention discloses a glucose isomerase, an encoding gene, a recombinant vector, a genetically engineered bacteria and a mutant thereof and an application of the glucose isomerase and the mutant thereof in preparation of D-fructose for catalyzing D-glucose isomerization. A recombinant escherichia coli capable of efficiently expressing high-temperature resistant glucose isomerase, provided by the invention, solves the problem of low optimal reaction temperature of a common glucose isomerase, is applied to production of the glucose isomerase and has the advantages of high yield, simple process and convenient industrial application; the strain can be directly used for the high fructose syrup production without cell disruption; after finishing fermentation, the total enzyme activity for D-fructose at the temperature of 90 DEG C is 414.3U / g, the conversion rate of D-fructose is 55.4%, the remnant enzyme activity at the temperature of 90 DEG C after storage for 24h is 68% and the equilibrium of reaction is shortened to 1.5h; a good technical support for the large-scale production of the high fructose syrup and for the use of high fructose syrup as an important food additive can be provided.
Owner:ZHEJIANG UNIV OF TECH

Recombinant escherichia coli for synthesizing lactoyl N-neotetraose and construction method and application of recombinant escherichia coli

The invention discloses recombinant escherichia coli for synthesizing lactoyl N-neotetraose and a construction method and application of the recombinant escherichia coli. The recombinant escherichia coli are obtained by knocking out a beta-galactosidase lacZ gene, a glucosamine-6-phosphodeaminase nagB gene, a UDP-acetylglucosamine epitope isomerase wecB gene and a UDP-glucose dehydrogenase ugd gene in the host genome of escherichia coli and overexpressing a lactose transportase lacY gene, beta-1, 3-N-glucosaminidase lgtA gene and a beta-1, 4 galactosyltransferase lgtB gene on the genome. The yield of lactoyl-N-neotetraose synthesized by recombinant escherichia coli reaches 985 mg / L, and a foundation is laid for further metabolic engineering transformation of escherichia coli to produce lactoyl-N-neotetraose.
Owner:JIANGNAN UNIV

Chondroitin producing genetic engineering bacterium, method for constructing same and application of chondroitin producing genetic engineering bacterium

The invention discloses a chondroitin producing genetic engineering bacterium, a method for constructing the same and application of the chondroitin producing genetic engineering bacterium, and belongs to the technical field of genetic engineering. Chondroitin synthetase genes and UDP (uridine diphosphate)-glucosamine isomerase genes are transferred into corynebacterium glutamicum to obtain the chondroitin producing genetic engineering bacterium. The chondroitin producing genetic engineering bacterium, the method and the application have the advantages that the corynebacterium glutamicum is used as a host for the chondroitin producing genetic engineering bacterium, is a GRAS (generally recognized as safe) strain affirmed by the Food and Drug Administration of the America, does not secreteoptional endotoxin or exotoxin, is safe and can be applied to producing amino acid or food additives and the like for a long term, and chondroitin produced by the chondroitin producing genetic engineering bacterium is high in molecular weight and good in yield; the chondroitin yield of preferable recombinant bacteria of the chondroitin producing genetic engineering bacterium can reach 3.7 g / L, chondroitin products with high molecular weights can be produced by the chondroitin producing genetic engineering bacterium, and accordingly the chondroitin producing genetic engineering bacterium has anexcellent industrial prospect and can be effectively applied to medicines and health protection.
Owner:TSINGHUA UNIV

Glucose isomerase genes, encoding enzymes, vectors and engineering bacteria and application

InactiveCN104293759AIncrease productionEfficient expression resistant to high yieldBacteriaMicroorganism based processesEscherichia coliBiotechnology
The invention discloses glucose isomerase genes, encoding enzymes, recombinant vectors and genetic engineering bacteria and application of glucose isomerase gene engineering bacteria in aspects of preparation of D-fructose by virtue of catalysis of D-glucose isomerization. According to the recombinant Escherichia coli for efficiently expressing high-temperature-resistant glucose isomerase, the problem that general glucose isomerase is low in optimal reaction temperature is solved; the glucose isomerase produced by using the strain has the advantages of high yield, simple process, convenient industrial application and the like, the strain can be directly used for producing high fructose syrup, and cell disruption is not needed; and moreover, after fermentation is ended, the total enzyme activity of D-glucose reaches 715U / L under the condition of 85 DEG C, and the conversion rate of the D-fructose is 55 percent, so that a good technical support is provided for performing large-scale production of the high fructose syrup and taking the high fructose syrup as an important food additive.
Owner:ZHEJIANG UNIV OF TECH +1

Glucose isomerase mutant and application thereof

The invention discloses a glucose isomerase mutant and application thereof, belonging to the technical field of enzyme genetic engineering and enzyme engineering. According to the invention, a high temperature glucose isomerase gene (NCBI coding: CP000088) is obtained from total DNA of thermobifida fusca, and after site-directed mutagenesis, high efficiency expression of the glucose isomerase gene with a high conversion rate is realized, with the plasmid pT7-7 or a vector that can express glucose isomerase as an expression vector and E. coli BL21 (DE3) or a bacterial strain that can express glucose isomerase as an expression host; the glucose isomerase gene has altogether 1158 nucleotide and encodes 385 amino acids; expression plasmid is constructed in the invention, glucose isomerase is expressed through conversion of bacteria or yeast, and an obtained recombinase mutant has activity of glucose isomerase and has a conversion rate of 60% at a temperature of 70 DEG C, 7% higher than the conversion rate of a parent; optimum temperature of the recombinant glucose isomerase is 80 DEG C, an optimum pH is 10, and the half life of the recombinant glucose isomerase at a temperature of 70 DEG C is no less than 30 h. The recombinant glucose isomerase is particularly applicable to production of F55 high-fructose syrup in the industry of foodstuffs.
Owner:JIANGNAN UNIV

Construction method for streptomycete expression plasmids and production method for keratinase

The invention relates to a construction method for streptomycete expression plasmids and a production method for keratinase. The construction method for streptomycete expression plasmids includes the following steps: a promoter Xi of actinoplanes missouriensis xylose isomerase and a terminator of streptomycete avermitilis amylase are sleeved, and the promoter-shine-dalgarno (SD) sequence comes from 90 to 269bp of an actinoplanes missouriensis xylose isomerase gene; a cloned src family kinases (sfks) gene is inserted in a construction expression frame structure of a synthetic Xi promoter-SD-amyA2 terminator fragment, and then a conventional method is used for constructing the streptomycete expression plasmids. The production method for keratinase includes: leading the streptomycete expression plasmids into an expression host of streptomycete lividans TK24 through conjugal transfer, performing recombinant expression and generating the keratinase. Specific activity of a crude enzyme solution is 1700 U/mg after expression of the expression plasmids in the streptomycete lividans TK24 and is improved by 50 times compared with specific activity of starting strain streptomycete fradiae varieties S-221, and yield of the keratinase is much higher than starting strains after the expression.
Owner:SHANGHAI JIAO TONG UNIV

Disulfide bond isomerase gene Trpdi2 from Trichoderma reesei and application thereof

InactiveCN104694560AActivity recovery or maintenanceIncrease productionIsomerasesFermentationBiotechnologyNucleotide
The invention discloses a disulfide bond isomerase gene Trpdi2 from Trichoderma reesei and application thereof. The disulfide bond isomerase gene separated from Trichoderma reesei has a polynucleotide sequence of (a) (b) or (c): (a) a polynucleotide sequence shown as SEQ ID No.1; (b) a polynucleotide sequence complementary with the polynucleotide sequence in (a) according to the principle of complementary base pairing; and (c) a cDNA sequence of the polynucleotide sequence (a)shown in the SEQ ID NO.1 or (b) polynucleotide sequence: a polynucleotide sequence shown in SEQ ID NO.2. The invention also obtains disulfide isomerase the from the encoding of the disulfide bond isomerase gene separated from the Trichoderma reesei and verifies the application of disulfide isomerase.
Owner:TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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