1313 results about "Reductase" patented technology

Provided herein are methods of treatment comprising administering a selective ribonucleotide reductase (RNR) inhibitor in combination with other therapeutic agents, thereby treating cancer and preventing or reversing resistance to the therapeutic agents.

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 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.

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 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 present invention relates to a recombinant yeast with high squalene production, a construction method and a use thereof, wherein the recombinant yeast is a saccharomyces cerevisiae that expresses or overexpresses HMGR gene, ADA gene, LEU2 gene, MVA pathway gene and squalene synthase gene ERG9. The present invention integrates acetaldehyde dehydrogenase gene ADA and HMG-CoA reductase gene tHMGR in multiple copies in saccharomyces cerevisiae, integrates the screened high-activity phosphoketolase gene PK and phosphotransacetylase gene PTA, weakens epoxysqualene synthesis, strengthens the mevalonate pathway, overexpresses yeast endogenous squalene synthase gene, etc. to obtain a recombinant yeast with high squalene production, so that the production in the shake flask stage of the production process exceeds 5g / L, and the production in the fed batch fermentation stage exceeds 50g / L, breaking through the current bottleneck of microbial synthesis of squalene production, and is suitable for practical promotion and application.

The invention discloses a compound enzyme preparation for improving the absorption and utilization rate of a fertilizer and a preparation method thereof, and belongs to the technical field of enzyme preparations. The compound enzyme preparation is prepared from the following components in parts by weight: 100 parts of purified water, 6 to 8 parts of modified cellulase, 10 to 12 parts of immobilized protease, 10 to 12 parts of phosphatase, 10 to 14 parts of urease, 8 to 10 parts of nitrate reductase, 10 to 12 parts of pectinase, 8 to 10 parts of chitinase, 6 to 8 parts of xylanase, 10 to 12 parts of beta-glucosidase, 10 to 12 parts of superoxide dismutase and 4.5 to 6 parts of a microbial agent, the modified cellulase is obtained by chemically modifying cellulase; the immobilized protease is prepared by the following steps: adsorbing activated and coupled modified polylactic acid and a protease solution, crosslinking with glutaraldehyde, filtering, washing and drying. The compound enzyme preparation improves the fertilizer absorption and utilization rate of plants.

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 belongs to the field of agricultural biotechnology and environmental microbiological engineering, and discloses mesotrione nitroreductase Mstnr as well as a coding gene and application thereof. The nucleotide sequence of the gene is SEQ ID NO.1, the full length is 678bp, 225 amino acids are encoded, and the amino acid sequence of the gene is SEQ ID NO.2. The highest homology of the Mtnr and reported nitroreductase is only 49.2%, so that the Mtnr is a new nitroreductase gene. The nitroreductase Mtnr provided by the invention can catalyze nitro reduction of mesotrione and convert mesotrione into a product 2-amino-4-methylsulfonylbenzoic acid (AMBA) without herbicidal activity, which indicates that the Mtnr gene has a huge application prospect in biodegradation detoxification and repair of mesotrione.

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.

The invention belongs to the field of agricultural biotechnology and environmental microbiological engineering, and discloses mesotrione nitroreductase Mtnr as well as a coding gene and application thereof. The nucleotide sequence of the gene is SEQ ID NO.1, the full length is 756bp, 251 amino acids are encoded, and the amino acid sequence is SEQ ID NO.2. The maximum homology of the Mstnr and reported nitroreductase is only 50.6%, so that the Mstnr is a new nitroreductase gene. The nitroreductase Mstnr provided by the invention can catalyze nitro reduction of mesotrione, and the mesotrione is converted into a product 2-amino-4-methylsulfonyl benzoic acid (AMBA) without herbicidal activity, so that the Mstnr gene has a huge application prospect in biodegradation detoxification and repair of the mesotrione.

The invention discloses a thermostable conjugated polyketoreductase mutant, which is obtained by performing single-point mutation or multi-point combined mutation on the 25th site, the 92th site, the 214th site or the 256th site of an amino acid sequence as shown in SEQ ID NO.1. The invention further discloses a preparation method of the thermostable conjugated polyketoreductase mutant. The initial enzyme activity of the mutant is high, and the residual enzyme activity is gt after incubation at 50 DEG C; the Tm value can be increased by more than 9 DEG C compared with that of an original strain, the 50 DEG C half-life period t1 / 2 is increased by more than 377.8 times, and the thermal stability is obviously improved. The invention also provides a coding gene, a recombinant vector containing the coding gene and a co-expression engineering bacterium. The thermostable conjugated polyketoreductase mutant, glucose dehydrogenase and L-pantolactone dehydrogenase are used for constructing a three-enzyme co-expression engineering bacterium as a biocatalyst, racemization pantolactone is catalyzed to synthesize DPL, for a 1M substrate, the yield of the DPL reaches 97.5% within 36 hours, KPL hydrolysis is avoided, the reaction time is shorter, the product concentration is higher, and the thermostable conjugated polyketoreductase mutant has great market application value.

The invention belongs to the field of agricultural biotechnology and environmental microbiological engineering, and discloses a novel mesotrione nitroreductase Bamnr, and a coding gene and application thereof. The nucleotide sequence of the gene is SEQ ID NO.1, the full length is 639bp, 212 amino acids are encoded, and the amino acid sequence of the gene is SEQ ID NO.2. The highest homology of Bamnr and reported nitroreductase is only 69.4%, so that Bamnr is a new nitroreductase gene. The nitroreductase Bamnr provided by the invention can catalyze nitro reduction of mesotrione, and the mesotrione is converted into a product 2-amino-4-methylsulfonyl benzoic acid (AMBA) without herbicidal activity, so that the Bamnr gene has a huge application prospect in biodegradation detoxification and repair of the mesotrione.

The invention belongs to the field of agricultural biotechnology and environmental microbiological engineering, and discloses a herbicide mesotrione degradation detoxification enzyme protein, and a coding gene and application thereof. The nucleotide sequence of the gene is SEQ ID NO.1, the full length of the gene is 738bp, 245 amino acids are encoded, and the amino acid sequence of the gene is SEQ ID NO.2. The maximum homology of the Mdmnr and reported nitroreductase is only 48%, so that the Mdmnr is a new nitroreductase gene. The nitroreductase Mdmnr provided by the invention can catalyze nitro reduction of mesotrione, and the mesotrione is converted into a product AMBA without herbicidal activity, which indicates that the Mdmnr gene has huge application prospects in mesotrione-resistant transgenic engineering and biodegradation detoxification and repair of mesotrione residual pollution.

The invention relates to the technical field of microbial genetic engineering, in particular to a method for improving cordycepin synthesized by saccharomyces cerevisiae and saccharomyces cerevisiae engineering bacteria. According to the invention, codon-optimized 2 '-carbonyl-3'-deoxyadenosine reductase gene and 3 '-adenosine monophosphate phosphohydrolase gene from cordyceps militaris are expressed by using plasmids, and then the plasmids are transformed into host cells to obtain the saccharomyces cerevisiae engineering bacteria. And inoculating a seed solution obtained by seed culture of the activated saccharomyces cerevisiae engineering bacteria into a fermentation culture medium containing a metabolic effector, and carrying out fermentation culture to synthesize cordycepin. Metabolic effectors (Cu < 2 + >, Fe < 2 + >, Mg < 2 + >, Zn < 2 + >, citric acid, cysteine, aspartic acid, glycine, VB1 and tea polyphenol) are added into a fermentation culture medium strain, and the variety and concentration of the metabolic effectors are optimized, so that the yield and synthesis efficiency of cordycepin are remarkably improved. By using the combination of the strain obtained by the invention and the optimal effector, the yield of cordycepin in a fermentation tank reaches 2.9 g / L.

The imine reductase mutant provided by the invention can catalyze asymmetric reductive amination of ethyl 3-formylhexanoate and (S)-2-aminobutanamide to synthesize brivaracetam, two chiral centers of brivaracetam are constructed in one step, and the problems of poor environmental protection property, low atom utilization rate, long route and the like in the existing industrial synthesis route are solved. Wide application prospects are realized.

Disclosed are, inter alia, double stranded RNAi (dsRNAi) agents inhibiting expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), for example, human HMGCR, compositions including the same, and methods of treatment using the same.

The invention discloses a heme oxygenase mutant and bilirubin efficiently synthesized by a multienzyme coupling system of the heme oxygenase mutant, and belongs to the field of biological catalysis engineering. The invention provides a heme oxygenase mutant which is used for catalyzing heme to synthesize biliverdin. Coupling biliverdin reductase to catalyze biliverdin to synthesize bilirubin; cytochrome P450 reductase is introduced to provide electrons; removing competitive binding of the by-product to the substrate heme by carbon monoxide dehydrogenase; formate dehydrogenase is used for in-situ regeneration of coenzyme, and bilirubin is efficiently synthesized through a one-pot method. According to the heme oxygenase mutant, the two core enzymes are coupled with three external assistant enzymes, recombinant escherichia coli is constructed, heme is catalyzed, exogenous NADPH does not need to be added, and the yield of bilirubin within 4 h reaches 1611 mg / L and reaches the bilirubin synthesis level which is reported at present and has the highest yield and the shortest time. According to the technology, resource limitation is relieved, the synthesis method is easy to operate and high in yield, and efficient and green synthesis of bilirubin is achieved.

The invention relates to the field of biochemical engineering, in particular to an alcohol dehydrogenase mutant and application thereof. The mutant is obtained by mutating alcohol dehydrogenase with an amino acid sequence encoding SEQ ID NO: 4 at one or more sites selected from 62nd, 108th, 130th, 132, 134, 182nd, 206th, 287th, 303th, 313th and 314, and the mutation is that the amino acid at the site is mutated into one of A, G, V, L, I, P, F, Y, W, S, T, C, M, N, Q, D, E, K, R and H. According to the alcohol dehydrogenase mutant and the application of the alcohol dehydrogenase mutant, the feeding amount of substrates is increased; the high conversion rate (gt; 99%) of the substrate is realized; according to the present invention, the new YqjM has the good product ee value (gt; 98%), and it is determined that the YqjM derived from the bacillus subtilis old yellow enzyme (belonging to ketene reductase) has the optimal catalytic activity, such that the good industrial application performance is provided.

The invention discloses a genetically engineered bacterium for producing uridine diphosphate glucose as well as a construction method and application of the genetically engineered bacterium, and belongs to the technical field of genetic engineering. The following genes are overexpressed in the genetically engineered bacterium: a pyrimidine nucleoside operon gene, a carbamyl phosphate synthetase gene, an aspartate carbamyl transaminase gene, an orotic acid phosphoribose transferase gene, a phosphoribose pyrophosphate synthase gene, a uridylate kinase gene and a glucophosphate mutase gene. A uridine triphosphate glucose-1-phosphate uridine group transferase gene; the genetically engineered bacterium does not express the following genes: a purine nucleotide repressor protein gene, a deoxyribonucleoside triphosphate reductase gene, a nucleoside triphosphate pyrophosphohydrolase gene, a cytidine triphosphate synthase gene, a pyrimidine nucleotidase gene, a nucleotidase gene, an ornithine carbamoyltransferase gene, a UDPG dehydrogenase gene and a lacI repressor protein gene. The genetically engineered bacterium can be used for efficiently synthesizing UDPG by taking glucose as a substrate.

The invention provides a recombinant pichia kudriavzevii strain and application thereof, pichia kudriavzevii is taken as an original strain, and a reduced TCA pathway is introduced and comprises the following steps: overexpressing fumarate reductase FRD in the strain; weakening or inactivating an ethanol pathway, comprising: knocking out any one or a combination of at least two of encoding genes of ethanol dehydrogenase ADH1, ADH3 or ADH6; enhancing the PPP pathway in the pentose phosphate pathway of the strain, including: overexpressing glucose-6-phosphate dehydrogenase ZWF1 and / or 6-phosphogluconate dehydrogenase GND1; the method for reducing the activity of the malate dehydrogenase MDH2 in the TCA circulation pathway comprises the step of knocking out the coding gene of the malate dehydrogenase MDH2 in the TCA pathway. The pichia kudriavzevii is modified by using genetic engineering and metabolic engineering methods, so that the yield and the yield of succinic acid are increased, and the pichia kudriavzevii has important application prospects in industrial production.

The invention discloses bacillus subtilis with high yield of surfactin, a related enzyme mutant and a preparation method of the bacillus subtilis, the bacillus subtilis is obtained by a gene modification method and can produce the surfactin with high yield, and the highest yield can reach 9.4 g / L; the invention also provides key enzyme mutants participating in surfactin synthesis in the bacillus subtilis, and the key enzyme mutants comprise a biotin carboxylase IdeHA mutant, a biotin carboxylase accBC mutant, an enoyl reductase FabI mutant, a malonyl transferase FabD mutant and a dihydrolipoyl dehydrogenase IPdU mutant. The key enzyme mutants comprise a biotin carboxylase IdeHA mutant, a biotin carboxylase accBC mutant, an enoyl reductase FabI mutant, a malonyl transferase FabD mutant and a dihydrolipoyl dehydrogenase IPdU mutant. The bacillus subtilis for high-yield surfactin and the preparation method thereof disclosed by the invention are beneficial to realizing industrial production and commercial application of the surfactin.

The invention belongs to the field of environmental microorganisms and agriculture, and discloses mesotrione nitroreductase as well as a coding gene and application thereof. The amino acid sequence of the mesotrione nitroreductase MnrA is as shown in SEQ ID NO. 2; the nucleotide sequence of the gene for coding mesotrione nitroreductase MnrA is as shown in SEQ ID NO. 1; the invention also provides application of the mesotrione nitroreductase and the gene as well as an expression box, a recombinant vector and a genetically engineered bacterium capable of degrading mesotrione. The homology of the nitroreductase MnrA provided by the invention and reported nitroreductase is lower than 20%, and the nitroreductase MnrA is a novel nitroreductase. The nitroreductase MnrA disclosed by the invention can catalyze nitro reduction of mesotrione, and the mesotrione is converted into a product AMBA without herbicidal activity, so that the nitroreductase MnrA and the coding gene thereof have huge application prospects in mesotrione-resistant transgenic engineering and biodegradation detoxification and repair of mesotrione residual pollution.

The invention relates to an application of a cytochrome P450 reductase 10273 inactivating agent in construction of an aspergillus terreus engineering strain for high yield of emodin or a derivative thereof. The invention further relates to aspergillus terreus engineering bacteria for high yield of emodin or the emodin derivatives and a construction method of the aspergillus terreus engineering bacteria. By knocking out the cytochrome P450 reductase gene 10273, the activation of anthraquinone methyl hydroxylase 4804 is blocked in aspergillus terreus, so that the hydroxylation pathway of the emodin is blocked, the purity of the emodin and the emodin-3-methyl ether in aspergillus terreus cells is improved, the production cost of the emodin and the emodin-3-methyl ether is reduced, and the method is suitable for industrial production. And a foundation is provided for the microbial industrial synthesis of the emodin and the emodin-3-methyl ether.

The invention relates to the technical field of biochemical engineering, in particular to a method for preparing chiral alpha-trifluoromethyl alcohol by using imine reductase AtRedAm, trifluoromethyl ketone is reduced into corresponding chiral trifluoromethyl alcohol at an excellent conversion rate by using the imine reductase AtRedAm, and enantioselectivity is high. The reaction depends on an NADPH (Nicotinamide Adenine Dinucleotide Phosphate) coenzyme circulation system, 10% dimethyl sulfoxide is used as a reaction cosolvent, the reaction is carried out in a 100mM (pH = 7.5) phosphate buffer solution for 16 hours, the highest molar conversion rate reaches 99%, and the highest optical purity reaches 95%.

Certain C. glutamicum strains overexpress genes coding for enzymes having the function of aspartate-semialdehyde dehydrogenase, aspartate aminotransferase, dihydrodipicolinate synthase, dihydrodipicolinate reductase, diaminopimelate decarboxylase, aspartatokinase and diaminopimelate dehydrogenase. The C. glutamicum strains also express NAD(P)(+) transhydrogenase subunit alpha PntA and NAD(P)(+) transhydrogenase subunit beta PntB of Corynebacterium urealyticum. A method is developed for the fermentative production of L-lysine using such C. glutamicum strains.