945 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.

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 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 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 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 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 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 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%.

The invention discloses a genetically engineered bacterium and an application of the genetically engineered bacterium in production of cis-3-hydroxy-L-proline. According to the genetically engineered bacterium, a proline dehydrogenase gene putA and a proline transporter gene putP in a starting bacterium are silenced, and meanwhile, a gamma-glutamyl kinase mutant gene proBD107N, a gamma-glutamyl phosphate reductase gene proA and a cis-3-hydroxy-L-proline hydroxylase gene P3H are subjected to overexpression. And by comparing a plurality of common escherichia coli, the cis-3-hydroxy-L-proline hydroxylase gene P3H and different gamma-glutamyl kinase mutant genes, the highest yield combination is preferably selected. The strain takes glucose as a raw material, so that the production cost is greatly saved, and the strain has a relatively good industrial utilization value.

The invention relates to the field of biological engineering and technology, in particular to an enzyme for synthesizing glabridin, a biological synthesis method of glabridin and a bacterial strain. The invention discloses a series of glabridin synthesizing enzymes (phenylalanine ammonialyase, cinnamic acid-4-hydrogenase, tyrosine ammonialyase, p-coumaric acid coenzyme A ligase, chalcone synthase, chalcone reductase, chalcone isomerase, isoflavone synthase, 2-hydroxyisoflavone dehydratase, 4 '-oxymethyltransferase, 4'-hydroxymethyltransferase and 4 '-hydroxymethyltransferase) derived from glycyrrhiza glabra for the first time. ) can be used in the preparation of a compound (e.g., isoflavone 2 '-hydrogenase, isoflavone reductase, Vissisoketoreductase, isoprenyltransferase, pterocarpin reductase, oxidative cyclase, demethylase, cytochrome P450 reductase, and the like). The de novo synthesis of the glabridin in a microbial cell factory is realized for the first time, and the application prospect of the glabridin biological manufacturing industry is greatly promoted.

The invention discloses a recombinant strain for producing squalene as well as a construction method and application of the recombinant strain. The construction method comprises the following steps: heterologously expressing cDNA (complementary deoxyribonucleic acid) of each gene in a squalene synthesis route from filamentous fungi in yeast; the squalene synthetic pathway genes comprise a mevalonic acid-5-phosphate kinase gene, a squalene synthase gene, an acetoacetyl coenzyme A thiolase gene, a mevalonic acid kinase gene, a hydroxymethyl glutaryl coenzyme a synthetase gene, a mevalonic acid pyrophosphate decarboxylase gene, a farnesyl pyrophosphate synthetase gene and isopentenyl pyrophosphate. A dimethallylpyrophosphate isomerase gene and a truncated hydroxymethyl glutaryl coenzyme a reductase gene; the culture medium is characterized in that the filamentous fungi are Saroocladium oryzae or Stibella sp. 1056, and the culture medium is a culture medium. The excavated squalene synthesis route can complete efficient de novo synthesis of squalene, and the squalene yield is increased.

The invention belongs to the technical field of biology, and particularly relates to a method for producing xylitol through two-step enzyme catalysis of hemicellulose hydrolysate, and the method comprises the following steps: taking hemicellulose hydrolysate as a raw material, adding arabinose dehydrogenase, NADH oxidase and coenzyme NADH, and carrying out first-step fermentation until no L-arabinose residue exists in fermentation liquor to obtain fermentation reaction liquor I; and adding xylose reductase and glucose dehydrogenase into the fermentation reaction liquid I, and carrying out second-step fermentation to obtain a fermentation product containing xylitol. According to the two-step enzyme catalysis method provided by the invention, xylitol is prepared, and high-purity xylose does not need to be used as a raw material; according to the present invention, the cheap hemicellulose hydrolysate can be directly adopted as the raw material, the xylose conversion rate and the xylitol yield can exceed 99%, the impurity arabitol content in the product is low, the other impurity saccharic acid is easy to separate, and the problems of complex process, large separation difficulty and high cost in the xylitol refining process are successfully solved.

The invention discloses a dihydrodaidzein reductase mutant and application of the dihydrodaidzein reductase mutant in synthesis of (3S, 4R)-tetrahydrodaidzein, and belongs to the technical field of biology. According to the mutant, wild-type dihydrodaidzein reductase DHDR is taken as a parent, single-point mutation or combined mutation is carried out, and a DHDR mutant is obtained; the parent site of the single point mutation is one of the 120th site, the 122nd site, the 170th site, the 178th site, the 214th site, the 219th site, the 220th site or the 220th site; the combined mutation adopts two modes: one mode is that a single-point mutation DHDR mutant is taken as a parent, mutation is carried out, and a combined mutation mutant I is obtained; and 2, taking the DHDR combined mutation mutant I as a parent, and performing mutation to obtain a combined mutation mutant II. The DHDR mutant disclosed by the invention is used for synthesizing (3S, 4R)-tetrahydrodaidzein by reducing dihydrodaidzein, and asymmetric reduction with high activity and high stereoselectivity can be realized.

The present invention relates to a process for producing a chiral aromatic amine, said process comprising the steps of: a) providing an aromatic imine, and b) contacting said aromatic imine from step a) with an imine reductase, thereby stereoselectively reducing said aromatic imine to a chiral aromatic amine with said imine reductase; and to novel imine reductases capable of catalytic reactions.

The invention provides a lysine artificial operon and application thereof. The gene sequence of the artificial operon comprises a promoter, a lysC gene for coding aspartate kinase, an asd gene for coding aspartate semialdehyde dehydrogenase, a dapA gene for coding dihydropicolinic acid synthetase and a dapB gene for coding dihydropicolinic acid reductase, the ddh gene is used for coding diaminopimelate dehydrogenase; the lysA gene is used for coding diaminopimelate decarboxylase; except for the first lysC gene, sequences of RBS ribose binding sites of other genes are RBS ribose binding sites translated after gene transcription by intercepting 20-26nt DNA sequences at the upstream of a coding region ATG (adenosine triphosphate); the RBS ribose binding site is connected in front of the corresponding gene.

The invention relates to the field of biological engineering and technology, in particular to an enzyme for synthesizing glabridin, a biological synthesis method of glabridin and an engineering-modified biological host. The invention discloses a series of glabridin synthesizing enzymes (4 '-O-methyltransferase, isoflavone 2'-hydrogenase, isoflavone reductase, Vissisoketoreductase, isoprenyltransferase, pterocarpin reductase, oxidative cyclase and demethylase) from glycyrrhiza glabra for the first time, and the glabridin synthesizing enzymes have corresponding activities in various biological hosts. The heterologous biosynthesis of the glabridin is realized for the first time, and the yield of the glabridin can reach 60% or above.

The present invention relates to a mutant ketoreductase, a nucleic acid encoding the mutant ketoreductase, a vector comprising the nucleic acid, a method of enzymatic reduction of a prochiral ketone with the mutant ketoreductase and formation of a chiral alcohol, the use of said mutant ketoreductase for the preparation of chiral alcohols and the use of said method for the preparation of pharmaceutically active morpholine compounds.

The present disclosure relates to recombinant microorganisms, such as bacteria, e.g., E. coli, engineered to have improved flux and / or yield of the DXP pathway. Cells can be engineered to comprise nucleotide sequences comprising dxs polypeptides (EC 2.2.1.7); dxr polypeptides (EC 1.1.1.267); ispD polypeptides (EC 2.7.7.60); ispE polypeptides (EC 2.7.1.148); ispF polypeptides (EC 4.6.1.12); ispG polypeptides (EC 1.17.7.1 or EC 1.17.7.3); ispH polypeptides (EC 1.17.7.4); and idi polypeptides (EC 5.3.3.2). Cells can be further engineered to express redox polypeptides, e.g., ferredoxins, flavodoxins, and / or flavodoxin / ferredoxin--NADP reductases (EC 1.19.1.1 or EC 1.18.1.2), e.g., to increase the activity of the ispG and / or ispH polypeptides. Some or all of the nucleotide sequences can be heterologous to the cells. The recombinant microorganisms can be used in methods of isoprenoid production.