13675results about "Oxidoreductases" patented technology

Disclosed herein are methods, compositions, and kits for determining the presence or absence of a target polypeptide (e.g., a polypeptide involved in nitrogen fixation) in a sample (e.g., a sample of soil from the rhizosphere of a non-leguminous plant).

The invention belongs to the technical field of bioengineering, and discloses a recombinant yarrowia lipolytica capable of efficiently and rapidly producing sclareol, which is characterized in that optimized sclareol synthase related coding genes ScLPPS, ScSCS, GGPPS1, GGPPS2, ERG20 and tHMGR coding genes are inserted into a yarrowia lipolytica genome; inserting optimized pentadienol utilization pathway coding genes such as IPK, THIM and IDI coding genes; and inserting an optimized ATP: citrate lyase coding gene ACLY to obtain the ATP: citrate lyase coding gene ACLY. The recombinant yarrowia lipolytica is based on knockout of a yarrowia lipolytica strain responsible for coding a non-homologous recombination gene ku70, so that the homologous recombination capability of the yarrowia lipolytica strain is enhanced, gene integration is realized through the homologous recombination function of the yarrowia lipolytica, the genetic stability of the introduced gene can be greatly improved, and the recombinant yarrowia lipolytica strain has a good application prospect. The construction method of the recombinant yarrowia lipolytica strain is efficient and simple to operate.

Methods for making synthetic gene clusters are described.

The invention relates to a recombinant escherichia coli THRS and a method for synthesizing L-threonine by using the recombinant escherichia coli THRS. The invention belongs to the technical field of bioengineering, and provides a recombinant escherichia coli THRS which overexpresses encoding genes of ppc, aspC, aspA, thrAmutBC, asd, rhtC, rhtA, asdtm and thrAmut, knockout an encoding gene of fnr and knockout an encoding gene of iclR, the recombinant escherichia coli THRS takes glucose as a substrate for fermentation, and the yield and conversion rate of L-threonine are obviously improved; the L-threonine strain has the advantages that the L-threonine strain does not contain exogenous plasmids, all modification sites are integrated onto a genome, higher stability is achieved, antibiotics do not need to be added into a fermentation culture medium of the strain, and the produced L-threonine product is free of antibiotic residues and higher in safety.

The invention relates to recombinant escherichia coli THRS-6 and a method for synthesizing L-threonine by using the recombinant escherichia coli THRS-6. The invention belongs to the technical field of bioengineering, and relates to an aspartic acid kinase thrAmut and pyridine nucleotide transhydrogenase pntAB gene derived from escherichia coli, an aspartic acid dehydrogenase aspDH gene derived from pseudomonas aeruginosa, a hemoglobin vgb gene derived from vitreoscilla and a transcriptional regulation factor irrE gene derived from radiococcus. The recombinant escherichia coli is used as a substrate, the ldhA gene is knocked out to further modify the recombinant escherichia coli, the recombinant escherichia coli THRS-6 with glucose as the substrate is obtained, the THRS-6 is used for fermentation, and the yield and conversion rate of the L-threonine are obviously improved.

The invention discloses construction and application of an engineering bacterium for co-expressing D-mannitol dehydrogenase and D-glucose dehydrogenase, and belongs to the technical field of genetic engineering. The technical problem to be solved by the invention is how to co-express D-mannitol dehydrogenase and D-glucose dehydrogenase through a biosynthetic pathway. In order to solve the technical problem, the invention provides recombinant escherichia coli. The recombinant escherichia coli is characterized by containing a D-mannitol dehydrogenase gene, a D-glucose dehydrogenase gene, a glucokinase gene, an acetyl coenzyme A synthetase gene, a D-lactose transporter gene, a D-glucose PTS permease-free gene and a pyruvate oxidase-free gene. And no regulator gene of galactose operon is contained. The invention establishes a preparation method for co-producing D-mannitol and sodium gluconate, which has guiding significance for the production of D-mannitol and sodium gluconate.

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.

The invention provides a CYP450 zymoprotein capable of catalyzing formation of neoandrographolide aglycone, a coding gene and application, and belongs to the field of biology. According to the present invention, bioinformatics analysis and differential expression analysis are performed on transcriptome data of different tissue parts, the gene ApCYP71BE50 capable of catalyzing (4R, 5S, 9R, 10S)-labda-8 (17), 13-dien-15, 19-diol to form the neoandrographolide aglycone is excavated, and it is verified that the CYP450 enzyme protein encoded by the gene ApCYP71BE50 can perform continuous oxidation on the C16 site of (4R, 5S, 9R, 10S)-labda-8 (17), 13-dien-15, 19-diol to form the neoandrographolide aglycone. The invention provides an important gene element for biosynthesis of andrographolide, and also provides a key gene locus for molecular design breeding of andrographis paniculata.

The invention belongs to the technical field of genetic engineering and metabolic engineering, and discloses an escherichia coli genetically engineered bacterium for fermentation production of L-histidine, a method and application of the escherichia coli genetically engineered bacterium for fermentation production of L-histidine. The escherichia coli genetically engineered bacterium is prepared by taking wild escherichia coli MG1655 as a starting strain, and knocking out a gene lacI and a gene purR on a genome; integrating the gene prs, the gene zwf and the gene gnd; the method comprises the following steps: integrating genes hisG, hisD, hisB, hisC, hisA, hisF / H and hisIE; and a gene cgl2458 is integrated. According to the invention, a molecular biological technology is utilized to integrate histidine operon in a serratia marcescens mutant strain with high histidine yield into an escherichia coli genome, and an escherichia coli genetically engineered strain with high histidine yield is obtained by enhancing precursor PRPP supply, histidine efflux and other strategies, so that the problems of thick fermentation liquor, high histidine yield and high histidine yield can be solved. And the method has a relatively good industrial application potential.

The present disclosure provides guided microbial remodeling (GMR) methods for the rational improvement of plant-associated microbes to perform plant-beneficial functions. The GMR methods described herein allow for non-intergeneric genetic optimization of key regulatory networks within the microbes, which improve plant-beneficial functions over wild-type microbes but don't have the risks associated with transgenic approaches (e.g., unpredictable gene function, public and regulatory concerns, etc.). The present disclosure also provides remodeled microbes and compositions thereof. The utilization of remodeled microbes and compositions thereof will enable farmers to realize more productive and predictable crop yields without the nutrient degradation, leaching, or toxic runoff associated with traditional synthetically derived fertilizers.

The invention belongs to the technical application field of microbial metabolic engineering and synthetic biology, and particularly relates to a pichia pastoris engineering strain for high-yield sclareol as well as a construction method and application of the pichia pastoris engineering strain. The method comprises the following steps: constructing a biosynthetic pathway of sclareol in a host strain, and optimizing an intracellular mevalonic acid metabolic pathway and a central metabolic pathway to obtain an engineering strain A; the host strain is pichia pastoris; or overexpressing or knocking out a metabolism regulation factor in the obtained engineering bacterium A to obtain an engineering bacterium strain B; or, in the obtained engineering bacterium B, a synthetic pathway is targeted to peroxisome and optimized by adopting a cell region compartment strategy, and an engineering bacterium strain C is obtained. The invention provides a pichia pastoris chassis cell for synthesizing a diterpenoid sclareol and an engineering strain for synthesizing sclareol. By introducing and optimizing the expression of a sclareol synthesis pathway, the fermentation yields of sclareol in shake flask batch and bioreactor batch fed-batch fermentation are respectively 631.6 mg / L and 10.5 g / L.

The present invention relates to transmissible elements (e.g. conjugative plasmids, phage and phagemid) which are engineered to express molecules of interest (MOIs) and a pathway modification system (PMS) which is able to introduce mutations in target endogenous genes in a recipient bacterium, or is able to produce nucleic acids which inhibit or increase expression of target endogenous genes in a recipient bacterium, or is able to express peptide inhibitor molecules which inhibit a target endogenous protein in a recipient bacterium. The invention relates to host cells containing the transmissible elements, to pharmaceutical compositions containing such transmissible elements or host cells, and their use in the treatment of various diseases.

The invention belongs to the technical field of biochemical engineering and the technical field of bioengineering, and discloses a schizochytrium limacinum engineering strain for producing EPA (eicosapentaenoic acid) by prolonging a desaturation path. Part or all of the extended desaturase genes from thraustochytrium sp. And fusarium sp. And saprolegnia are expressed. According to the invention, extended desaturase genes from various sources are integrated into a schizochytrium limacinum genome, so that a schizochytrium limacinum gene engineering strain efficiently expresses the extended desaturase genes, and a heterologous EPA synthesis path with high synthesis efficiency, short culture period, simple purification method and low cost is provided; a basis is provided for producing grease rich in EPA from schizochytrium limacinum, and the problems of non-sustainability, high cost and the like caused by obtaining EPA from fishes are solved.

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 mutant and a method for catalytically synthesizing high-concentration glassine by using the carbonyl reductase mutant, and belongs to the field of biological catalysis. According to the invention, a one-pot method is adopted to catalyze beta-acetone xyloside to synthesize high-concentration boine, carbonyl reductase CnMDR coded enzyme and rational design of a substrate pocket key are combined with multiple sites, beneficial mutations are combined, dominant mutant enzyme recombinant bacteria are used as a catalyst, beta-acetone xyloside is used as a substrate, and formate dehydrogenase variants and NAD + kinase are introduced at the same time, so that the high-concentration boine is synthesized. The NADP < + >-NADPH coenzyme is driven to circulate, the yield of the synthesized boine is 200g / L, and the conversion rate is gt; the ee value of the product is gt; the invention provides a one-pot efficient low-cost method for synthesizing the boine, only a trace amount (0.3 mM) of coenzyme needs to be added, byproducts in a reaction system are easy to separate, and great convenience is provided for industrial large-scale production.

The invention relates to the technical field of microbial fermentation, and discloses a genetically engineered bacterium of high-yield D-pantothenic acid as well as a preparation method and application of the genetically engineered bacterium. The D-PA yield of the engineering bacteria is greatly improved through the following effects that acetolactate synthase IlvB and IlvN shunted by pyruvic acid are subjected to overexpression, meanwhile, a transcription attenuation region is deleted, and valine feedback inhibition sites are mutated; the method comprises the following steps: overexpressing keto-alcohol acid reductoisomerase IlvC and dihydroxy acid dehydratase IlvD, and converting the IlvC coenzyme preference from NADPH into NADH, so as to break through the upstream of D-PA synthesis; overexpression of PanB is carried out, overexpression of a serine-glycine transport system is carried out, and synthesis of 5, 10-methylenetetrahydrofolic acid is promoted; meanwhile, the expression of ketopantoic acid reductase PanE is enhanced; and finally, over-expressing pantothenic acid synthetase PanC, and increasing the content of a cofactor ATP required by the PanC. According to the engineering bacteria provided by the invention, through overall reinforcement of the multiple modules, cell growth is improved due to balance of cofactors and increase of D-PA, and the yield of D-PA reaches an unprecedented height.

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.

Engineered CRISPR-Cas9 nucleases with altered and improved PAM specificities and their use in genomic engineering, epigenomic engineering, and genome targeting.

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 thermal-stability glucose oxidase mutant, polynucleotide for encoding the glucose oxidase mutant, a recombinant expression vector containing the polynucleotide, a host cell for expressing the glucose oxidase mutant, and applications of the thermal-stability glucose oxidase mutant, the polynucleotide, the recombinant expression vector and the host cell in the fields of food, chemical industry, medicine, agriculture or feed.

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.