9863results about "Oxidoreductases" patented technology

The present application relates to a kind of deletion mutant nucleic acid and its application in anti-herbicide.The deletion mutation occurs in the promoter of rice OsHPPD Gene, the length of deletion sequence is at least 10 bp, and the A in the start codon ATG of gene is 0, at least the bases located in the upstream of start codon ATG from 2041 to 2032 are deleted. OsHPPD The present application finds that by deleting part of the sequence in the promoter of rice OsHPPD Gene, rice can obtain resistance to HPPD inhibitor herbicides, which is of great application value for ensuring agricultural production safety and improving the efficiency of herbicide use.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P)+) or derivative thereof, an NAD(P)+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P)+) or derivative thereof, an NAD(P)+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

The present disclosure provides proteins useful for preventing, treating, inhibiting and / or eliminating infestation and infection of plants caused by various phytopathogenic pests, as well as formulations comprising such proteins, polynucleotides encoding such proteins, organisms expressing such proteins, and methods for preparing such proteins. And methods of using such proteins, formulations, polynucleotides, and organisms in agricultural and other effort areas.

The invention discloses acid-resistant pichia kudriavzevii and an application of the acid-resistant pichia kudriavzevii. The acid-resistant pichia kudriavzevii is Pichia kudriavzevii S9 and is preserved in the Guangdong Microbial Culture Collection Center on September 20, 2024, and the preservation number of the acid-resistant pichia kudriavzevii S9 is GDMCC.No: 65148. The acid-resistant pichia kudriavzevii S9 has the advantages that the acid-resistant pichia kudriavzevii S9 is used for According to the invention, a low-pH-resistant pichia kudriavzevii strain is screened from surface microorganisms of grapes, and the pichia kudriavzevii strain can tolerate an environment with pH of 1.8 and ferment organic acid under the condition of not adding a neutralizing agent; besides, genetic modification is carried out based on pichia kudriavzevii, including knocking out succinic acid degradation related genes, knocking out ethanol and glycerol production pathways and enhancing succinic acid transporter expression, so that engineering bacteria for high-yield succinic acid are obtained, and the fermentation yield of succinic acid reaches 31.74 g / L under the condition that a neutralizer is not added.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P)+) or derivative thereof; an NAD(P)+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

The invention provides a genetically engineered bacterium, a preparation method thereof and application of the genetically engineered bacterium in synthesis of sclareol. The genetically engineered bacterium comprises a CcLPPS gene derived from Cistanus crenatus, and a CcLPPS gene derived from Cistanus crenatus, the genetically engineered bacterium is selected from yarrowia lipolytica. According to the invention, the stably expressed sclareol efficient de novo synthesis strain is constructed, and no organic reagent or any path intermediate substance is added in the whole fermentation process, so that the production cost is greatly reduced, and the subsequent separation and purification steps are simplified. The engineering yarrowia lipolytica strain provided by the invention is high in production intensity and simple in fermentation process, has a relatively good application prospect, and lays a foundation for producing sclareol by a microbial fermentation method.

The invention provides a carbonyl reductase mutant. The carbonyl reductase mutant is obtained by performing single-point mutation or multi-point combined mutation on the 17th site, the 40th site and the 64th site of an amino acid sequence as shown in SEQ ID NO.1. The invention also provides a coding gene, a recombinant vector containing the coding gene, a co-expression engineering bacterium and an application of the co-expression engineering bacterium. Compared with a wild type enzyme, the carbonyl reductase mutant has high activity and high stereoselectivity, the enzyme activity can reach more than two times that of the wild type enzyme, the catalytic efficiency on a substrate precursor ketone is remarkably improved, the yield of a statin drug intermediate and chiral alcohol synthesized by an enzyme method of the carbonyl reductase mutant is remarkably improved, and the yield of a rosuvastatin intermediate (3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R) is remarkably improved. The highest yields of the (3R, 5R)-6-chloro-3, 5-dihydroxyhexanoic acid tert-butyl ester and the atorvastatin intermediate (3R, 5R)-6-cyano-3, 5-dihydroxyhexanoic acid tert-butyl ester reach 99% and 97.6% respectively, e.e. Is larger than 99%, and the method has high industrial application value.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P)+) or derivative thereof; an NAD(P)+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

The invention relates to an engineering bacterium of high-yield ornamental blue as well as a construction method and application of the engineering bacterium, and belongs to the technical field of natural dye synthesis by a biological method. The engineering bacterium capable of highly producing the ornamental blue, disclosed by the invention, expresses icd, bpsA, glnAY405F and gd hA; acnR, yggB, glsK, aceA and ldh are knocked out from the engineering bacteria. According to the application, yggB and aceA genes are knocked out, site-directed mutagenesis (Y405F) is carried out on a glnA gene, a glsK gene is replaced by a glnAY405F gene, an acnR gene is replaced by an icd gene, an ldh gene is replaced by a gdhA gene, and a bpsA gene is integrated, so that the formation of by-products such as lactate and succinic acid in a blue-viewing synthesis process can be blocked; the flow direction of a passage from citric acid to isocitric acid to alpha-ketoglutaric acid to glutamic acid to glutamine is improved, so that the concentration of glutamine in cells is improved, and the yield of the ornamental blue is improved.

The invention discloses a biosynthesis system for high-yield 1, 6-hexamethylenediamine and application of the biosynthesis system, and belongs to the technical field of biology. According to the method, key genes which are excavated and optimized in a 1, 6-hexamethylenediamine biosynthetic pathway are co-expressed in a single cell module or a multi-cell module, and different enzyme elements expressed by the cell module are subjected to enzyme cascade reaction, so that 1, 6-hexamethylenediamine can be efficiently synthesized by taking caprolactam or 6-aminocaproic acid as a substrate; furthermore, after the carboxylic acid reductase MaCAR is mutated, the catalytic activity of the carboxylic acid reductase MaCAR can be remarkably improved, and the yield of the 1, 6-hexamethylenediamine is further improved; in addition, the biosynthesis system disclosed by the invention directly takes the cell module as a whole-cell catalyst, and an enzyme purification step is avoided, so that the biosynthesis system has the advantage of low cost.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P)+) or derivative thereof, an NAD(P)+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P)+) or derivative thereof; an NAD(P)+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

The invention discloses a carbonyl reductase, a mutant and an application of the carbonyl reductase in synthesis of a chiral alcohol compound. The carbonyl reductase mutant is obtained by performing single-site or multi-site mutation on the 96th site, the 145th site, the 202nd site or the 206th site of the amino acid sequence of the carbonyl reductase LbCR derived from lactobacillus brevis as shown in SEQ ID No.2. The activity and stability of the carbonyl reductase LbCR mutant are obviously improved, and the carbonyl reductase LbCR mutant has a wide substrate spectrum. Compared with a wild type, the mutant LbCRM4 has the advantages that the enzyme activity is improved by 86.3 times, the substrate conversion rate is greater than 99%, the e.e.p value of the product is always kept at 99.5% or above, the space time yield reaches 1288.9 g / L / d, and the half-life period at 40 DEG C is 101.4 h and is improved by 83.5 times. The LbCR mutant has an industrial application prospect.

The invention relates to a saccharomyces cerevisiae strain for efficiently synthesizing cordycepin as well as a construction method and application of the saccharomyces cerevisiae strain, and belongs to the technical field of microbial genetic engineering. The construction method provided by the invention comprises the following steps: carrying out multi-copy genome integrated expression on heterologous codon optimized 2 '-carbonyl-3'-deoxyadenosine reductase genes and 3 '-adenosine monophosphate phosphohydrolase genes, so as to increase the dosage of key enzymes; and supplementing the defective gene of the engineering strain to obtain the saccharomyces cerevisiae strain B5U3 with high yield of cordycepin. According to the obtained saccharomyces cerevisiae strain for efficiently synthesizing the cordycepin, the cordycepin yield reaches 2.92 g / L after shake flask fermentation, the cordycepin yield reaches 7.5 g / L after fermentation in a fermentation tank, and a foundation is laid for safer and more reliable industrial production of the cordycepin.

The invention relates to the field of gene engineering, and discloses a D-3-phosphoglycerate dehydrogenase mutant, a coding gene thereof and application of the D-3-phosphoglycerate dehydrogenase mutant and the coding gene. Compared with a protein with an amino acid sequence as shown in SEQ ID NO: 1, the mutant comprises mutation at the 131 site and / or the 285 site. The mutant can specifically catalyze a substrate 3-phosphoglycerate to synthesize 3-phosphate-hydroxypyruvic acid, can effectively promote synthesis of L-serine in a metabolic pathway of microorganisms, improves the yield and conversion rate of the L-serine, and reduces the yield of byproducts at the same time.

The invention provides recombinant escherichia coli with high yield of ergothioneine and a method for preparing ergothioneine, the recombinant escherichia coli co-expresses ergothioneine anabolic enzymes Egt1 and Egt2, the methyltransferase structural domain of the Egt1 is derived from trichoderma reesei, mycobacterium smegmatis EgtD or rhizopus nigricans, and the methyltransferase structural domain of the Egt2 is derived from the trichoderma reesei, the mycobacterium smegmatis EgtD or the rhizopus nigricans. The histidine sulfoxide synthase structural domain is derived from trichoderma reesei, schizosaccharomyces pombe or rhizopus nigricans, and the Egt2 is derived from claviceps purpurea, neurospora crassa or aspergillus niger. After the recombinant escherichia coli obtained by screening is cultured for 68 hours in a 5L fermentation tank, the yield of ergothioneine can reach 5g / L or above; wherein after recombinant escherichia coli co-expressing Egt1 enzyme (the structural domain of methyltransferase is derived from mycobacterium smegmatis EgtD, and the structural domain of histidine sulfoxide synthase is derived from trichoderma reesei) and Egt2 enzyme (derived from neurospora crassa) are cultured in a 5L fermentation tank for 68 hours, the yield of ergothioneine can reach 8.7 g / L, and the production intensity reaches 127.9 mg / (L.h).

The invention discloses a formate dehydrogenase mutant and application thereof in catalytic synthesis of D-mannitol, and belongs to the technical field of bioengineering. According to the formate dehydrogenase mutant provided by the invention, the 18th-site glutamic acid of wild-type formate dehydrogenase with an amino acid sequence shown as SEQ ID NO.2 is mutated into proline, the 57th-site asparagine is mutated into glutamic acid, the 70th-site histidine is mutated into tryptophan, the 235th-site serine is mutated into threonine, and the 316th-site valine is mutated into threonine, so that the formate dehydrogenase mutant is high in enzyme activity and catalytic activity; the coenzyme circulation efficiency during the production of D-mannitol can be obviously improved, so that the yield of D-mannitol can be effectively improved. When the genetically engineered bacterium co-expressed by the formate dehydrogenase mutant and the mannitol dehydrogenase mutant is used for catalyzing 150 g / L fructose to produce D-mannitol, the yield of D-mannitol within 15 h can reach 140 g / L or above.

The invention discloses thermophilic fungus cleavable polysaccharide monooxygenase and application thereof, and belongs to the technical field of enzyme engineering and microbial engineering. The thermophilic fungus lysing polysaccharide monooxygenase provided by the invention has excellent thermal stability; when the lysable polysaccharide monooxygenase is used for synergistically hydrolyzing phosphoric acid swelling cellulose, microcrystalline cellulose and bagasse, the enzymolysis rate is increased, and the problems of poor thermal stability of the lysable polysaccharide monooxygenase, low enzymolysis rate of synergic cellulase and high LPMOs enzyme dosage are solved. The method has high practicability and can be widely applied to the enzymolysis process of various cellulosic substrates, so that the enzymolysis efficiency of the cellulosic substrates is remarkably improved.

The present invention provides a tobacco fragrance gene, its targeted sgRNA and applications, belonging to the technical field of tobacco breeding. A tobacco fragrance gene includes the NtBADH2a and / or NtBADH2b gene; the nucleotide sequence of the NtBADH2a gene is as shown in SEQ ID NO:1; the nucleotide sequence of the NtBADH2b gene is as shown in SEQ ID NO:2. The application of the tobacco fragrance gene as a target in improving the fragrance of tobacco. In view of the fact that the tobacco fragrance gene affects the content of tobacco fragrance substances through negative regulation, the present invention also provides the application of an sgRNA or gene editing system targeting the tobacco fragrance gene in constructing a tobacco line rich in fragrance substances. It can be seen that the tobacco fragrance gene provided by the present invention provides an effective target for tobacco breeding.

The invention discloses a method for preparing recombinant III-type human collagen. The method comprises the following steps: constructing genetically engineered bacteria for expressing proline hydroxylase and full-length recombinant III-type human collagen; and culturing the genetically engineered bacteria to obtain the recombinant III-type human collagen, the proline hydroxylase is prepared from P4Ha2, P4Hb and vP4H; the ratio of the copy number of the P4Ha2 to the copy number of the P4Hb to the copy number of the vP4H to the copy number of the full-length recombinant III type human collagen is (1.80 to 2.20) to (1.80 to 2.20) to (0.80 to 1.60) to 1. The method can be used for large-scale production, the purity is high, and the production cost can be reduced; the prepared recombinant III-type human collagen is completely consistent with the full-length sequence of natural III-type human collagen, the hydroxyproline content of the recombinant III-type human collagen is basically consistent with that of the natural III-type human collagen, and the recombinant III-type human collagen has a triple-helix structure, has the biological activity of the natural III-type human collagen, and is free of immunogenicity and virus hidden danger.

The invention relates to a gene modified cell and a method for producing fucosyllactose by using the gene modified cell, in particular to a kluyveromyces kluyveromyces gene modified cell and application thereof, and belongs to the technical field of biological gene engineering. The present invention provides a genetically modified cell comprising a recombinant nucleic acid sequence encoding a polypeptide having an [alpha]-1, 2-fucosyltransferase polypeptide ([alpha]-1, 2-fucosyltransferase) activity. The alpha-1, 2-fucosyltransferase polypeptide includes, but is not limited to, polypeptides having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical amino acid sequences with the amino acid sequences of SEQ ID NO: 1-25. According to the gene modified cell, the fucosyllactose synthesis capability of an original strain is improved.

The invention relates to the technical field of genetic engineering, and particularly discloses a genetic engineering bacterium for producing melanin as well as a construction method and application of the genetic engineering bacterium. According to the application, the yield of the melanin can be effectively increased by modifying a metabolic pathway of the genetically engineered bacterium, and the yield of the melanin reaches 6.2 g / L and 18.5 g / L respectively through shake-flask culture and 5L fermentation tank culture. Compared with the prior art, the yield of melanin produced by the genetically engineered bacterium is improved by 30% at the shake flask fermentation level, the fermentation yield of a fermentation tank is improved by 4.3 times, in addition, expensive tyrosine is changed into glucose as a fermentation precursor substance, the production cost is greatly reduced, and an efficient and feasible solution is provided for large-scale production of melanin.

The invention relates to the technical field of enzyme engineering, in particular to an L-glutamate oxidase mutant as well as preparation and application thereof. Through mutation of the 77th amino acid and the 171st amino acid, the active center configuration of the enzyme is optimized, the binding capacity with a substrate and the catalytic conversion efficiency are enhanced, and the enzyme activity is remarkably improved. The mutant disclosed by the invention shows stronger specificity to L-glutamic acid (salt), effectively reduces non-specific reaction and improves the product purity. And in the presence of a high-concentration substrate, the enzyme activity is not inhibited by the substrate. The prepared L-glutamate oxidase mutant with high enzymatic activity can be efficiently expressed in escherichia coli, the problem that the expression quantity of natural enzymes in streptomyces is limited is solved, the acquisition efficiency of the enzymes is greatly improved, and the production cost is reduced. The mutants can be directly applied to biological manufacturing of alpha-ketoglutaric acid, so that the production efficiency of products is remarkably improved.

The invention discloses a mesophilic carotenoid-producing recombinant yarrowia lipolytica strain as well as a construction method and application thereof, the recombinant strain exogenously expresses mesophilic original fatty acid desaturase (FAD3) and isocitrate dehydrogenase (IDH2) mutants from phaffia rhodozyma, and efficient synthesis under the condition of 30 DEG C is realized; secondly, a metabolic pathway of the recombinant strain is optimized based on a metabolic flow push-pull strategy, and isopentenyl pyrophosphate isomerase (IDI), farnesyl pyrophosphate synthase (ERG20), plant alkene dehydrogenase (CarB) and bifunctional plant alkene synthase (CarRP) are overexpressed, so that the flux of a precursor is increased; finally, gene multi-copy of astaxanthin synthase (crtS) and cytochrome P450 reductase (crtR) from phaffia rhodozyma is carried out, and the carotenoid production capacity of yarrowia lipolytica is further improved. The construction method of the mesophilic yarrowia lipolytica is simple, efficient synthesis of carotenoid can be better achieved at the higher temperature, the fermentation cost is greatly reduced, and the industrial production potential is achieved.

Methods for making synthetic gene clusters are described.

The invention discloses a method for efficiently synthesizing epiinositol by using a heat-resistant myo-inositol dehydrogenase mutant cascade system, and belongs to the field of biocatalysis engineering. Whole cells for expressing a scyllo-inositol dehydrogenase mutant and a heat-resistant scyllo-inositol dehydrogenase mutant are utilized to catalyze the scyllo-inositol to synthesize the epiinositol in a two-step cascade manner, and different from reported scyllo-inositol and D-chiral inositol synthesis methods, in the process of synthesizing the epiinositol by taking the scyllo-inositol as a substrate, the C4 position of the scyllo-inositol is dehydrogenated firstly, then the C4 position of the scyllo-inositol is dehydrogenated, and the C4 position of the D-chiral inositol According to the present invention, the structure of the substrate and the structure of the product are symmetric, and the enzyme is difficult to specifically recognize during the catalysis process, such that the synthesis of the different chiral compounds of the inositol is the important problem in the biological catalysis field. The invention provides a brand-new way for biocatalytically synthesizing the epiinositol, the yield of the epiinositol is increased to 1.89 g / L through enzyme modification, the conversion rate is 52.6%, and the yield and the conversion rate are the highest level reported publicly at present.

The invention provides a nano transdermal delivery system based on superoxide dismutase, the nano transdermal delivery system is of a capsule composite structure of a small molecule active component-superoxide dismutase-polymer shell layer, the small molecule active component is loaded in superoxide dismutase, the surface of the superoxide dismutase is coated by the polymer shell layer, and the superoxide dismutase is loaded in the capsule composite structure. The diameter of the superoxide dismutase-based active ingredient transdermal delivery system is 20-100 nm, and the thickness of the polymer shell layer is 7.5-52.5 nm. In the delivery system, the polymer shell layer provides a protection effect for the superoxide dismutase and the micromolecular functional substances with the anti-oxidation effect, the capsule composite structure is in a nanogel form, and the nanogel not only promotes the penetration depth of the capsule, but also reduces the irritation effect of the capsule on the skin. The raw materials are biologically friendly, the preparation method is simple, the yield is high, and industrial large-scale production can be realized.

A working wire of a continuous biological sensor is disclosed. The working wire includes a substrate for the sensor having a conductive surface. An enzyme layer is on the conductive surface and includes an enzyme, an immobilization matrix, and an enzyme immobilization network. The enzyme immobilization network is formed using a polymeric crosslinking agent and a non-polymeric crosslinking agent crosslinking the enzyme and the immobilization matrix. The polymeric crosslinking agent and the non-polymeric crosslinking agent are a combination of polyethylene glycol (PEG) dialdehyde and glutaraldehyde. A protective layer is over the enzyme layer.

The invention provides a keto-alcohol acid reductoisomerase mutant and application of the keto-alcohol acid reductoisomerase mutant in construction of genetically engineered bacteria for high yield of D-pantothenic acid. In order to solve the technical problem that in the prior art, the yield of D-pantothenic acid synthesized through microbial fermentation is not high, semi-rational design of enzyme is adopted, and a keto-alcohol acid reductoisomerase mutant with substrate specificity is obtained and applied to construction of genetically engineered bacteria for high yield of D-pantothenic acid. According to the invention, a genetically engineered bacterium with high yield of D-pantothenic acid is constructed by utilizing the keto-alcohol acid reductoisomerase mutant through a CRISPR-Cas9 gene editing technology, so that the yield of D-pantothenic acid is increased by about 12.04%, and compared with an original strain, the valine accumulation amount in obtained fermentation liquor is almost unchanged compared with that of a control, and the yield of D-pantothenic acid is greatly improved. However, the accumulation of branched chain amino acid isoleucine is less, so that the modification of the substrate specificity of the keto-alcohol acid reductoisomerase is achieved.