15 results about "Formate dehydrogenase" patented technology

The invention discloses a heme oxygenase mutant and application of the heme oxygenase mutant in preparation of bilirubin through multienzyme coupling, and belongs to the field of biological catalysis engineering. The forward mutant of the heme oxygenase is obtained through a directed evolution technology, and heme can be efficiently catalyzed to synthesize biliverdin; the bilirubin is synthesized by further introducing biliverdin reductase, coupling cytochrome P450 reductase and formate dehydrogenase, and coupling the two core enzymes with foreign assistant enzymes to construct recombinant escherichia coli to catalyze heme to synthesize bilirubin. In the system, NADPH does not need to be exogenously added, the reaction time is about 4 h, the yield of bilirubin reaches 2.10 g / L, accumulation of intermediate products is not observed in the catalysis process, and the highest yield and the shortest reaction time in bilirubin synthesis reported at home and abroad at present are achieved. Compared with the traditional pig bile extraction, the method disclosed by the invention breaks through the resource limitation, is simple to operate and high in yield, and realizes efficient and green synthesis of bilirubin.

The invention discloses hydrophilic super-macroporous polystyrene (HGPS (at) MAF-7 (at) FDH) composite microspheres loaded with an inorganic / organic hybrid cascade catalytic system and a preparation method of the hydrophilic super-macroporous polystyrene (HGPS (at) MAF-7 (at) FDH) composite microspheres. The method comprises the following steps: coupling HGPS microspheres with an organic ligand triazole through surface epoxy modification, and reacting the HGPS microspheres with a metal ion solution to obtain a metal ion-loaded microsphere skeleton; then dispersing the framework material in a buffer solution, adding an enzyme solution and an organic ligand triazole, and carrying out a one-pot reaction to obtain the HGPS-coated MAF-7-coated FDH composite microsphere which is used as an inorganic / organic hybrid cascade catalytic system to catalyze CO2 conversion to generate formic acid. According to the method, the defects that a nano material is easy to aggregate and collapse and difficult to recover are effectively overcome, and the material has high specific surface area, excellent mass transfer performance, biocompatibility and mechanical stability. The characteristics of the MAF-7 organic ligand and the controllable porous structure are beneficial to adsorption of CO2 gas, and the MAF-7 not only can be used as a nano-enzyme for replacing carbonic anhydrase to catalyze a CO2 hydration reaction, but also can be used as a carrier for immobilizing formate dehydrogenase. According to the HGPS and MAF-7 and FDH composite microspheres, the complexity of a multi-enzyme cascade reaction is simplified, the stability and robustness of enzymes are improved, a substrate channel is shortened, the overall reaction efficiency is improved, and the HGPS and MAF-7 and FDH composite microspheres have excellent popularization prospects in the fields of CO2 bioconversion and wider cascade reaction system biocatalysis.

This invention discloses a method for the enzymatic synthesis of L-aminobutyric acid (GABA), comprising the following steps: using threonine and ammonium formate as substrates, a three-enzyme combined catalytic reaction is carried out using leucine dehydrogenase, threonine deaminase, and formate dehydrogenase at a temperature of 40-55°C and a pH of 7.0-9.0 to obtain L-aminobutyric acid. The leucine dehydrogenase is the leucine dehydrogenase with the amino acid sequence shown in SEQ ID NO:1 or a mutant thereof. This invention can rapidly convert threonine to L-aminobutyric acid, yielding L-aminobutyric acid with high optical purity, thus improving economic efficiency.

The invention discloses a method for separating and preparing (R)-citronellal and co-producing (S)-citronellal, which takes raw materials containing (R)-citronellal and (S)-citronellal as substrates and carbonyl reductase and formate dehydrogenase as catalysts. According to the invention, co-production of (R)-citronellal and (S)-citronellal can be realized, and the (R)-citronellal and (S)-citronellal can be applied to the field of spices,

The invention discloses a method for synthesizing salicylic acid and derivatives thereof by multi-enzyme cascade catalysis of phenol, which comprises the following steps: reacting a reaction system containing phenol, enzyme, metal ion salt and bicarbonate to obtain salicylic acid and derivatives thereof; when the enzyme is carboxylase, the reaction system reacts to obtain salicylic acid; when the enzyme is a combination of carboxylase, carboxylate reductase, formate dehydrogenase and phosphokinase, the reaction system reacts to obtain salicylaldehyde; when the enzyme is a combination of carboxylase, carboxylate reductase, formate dehydrogenase, phosphokinase and alcohol dehydrogenase, the reaction system reacts to obtain salicyl alcohol; when the enzyme is a combination of carboxylase, carboxylate reductase, formate dehydrogenase, phosphokinase, alcohol dehydrogenase and glycosyltransferase, the reaction system reacts to obtain salicin. According to the method, a green synthesis path based on biological catalysis is utilized, phenol is converted into chemicals with higher economic value, and environmental pollution and carbon emission are reduced.

This invention discloses an enzymatically catalytic synthesis method for a chiral intermediate of dzoxadamine. Using methyl acetoacetate as a substrate, buffer, substrate, alcohol dehydrogenase, formate dehydrogenase, ammonium formate, and coenzyme are added to a reaction vessel. The reaction is carried out at 20–40°C for 2–24 hours. After extraction, separation, and rotary evaporation, the chiral intermediate (R)-3-hydroxybutyrate methyl ester is obtained. The alcohol dehydrogenase is a mutant of ADH1 derived from Rollstonella, and the amino acid sequence of ADH1 is shown in SEQ ID No. 1. This invention offers mild reaction conditions, high stereoselectivity, and environmental friendliness. It also boasts high conversion rate, high chiral purity, low enzyme dosage, tolerance to high substrate concentrations, and low preparation cost, making it suitable for industrial production. Furthermore, it avoids chiral resolution and heavy metal residues in the product, overcoming the shortcomings of chemical methods.

The invention discloses a preparation method of a sulfo-amino acid compound. The sulfo-amino acid compound is a compound as shown in a formula I, the method comprises the following steps: in the presence of a buffer solution, an ammonia source, nicotinamide adenine dinucleotide and formate dehydrogenase, catalyzing a thiomethyl alpha-butanone acid compound as shown in a formula II to perform asymmetric reductive amination reaction by using alpha-amino acid dehydrogenase containing an amino acid sequence as shown in SEQ ID NO: 1 or a mutant thereof, so as to generate the compound as shown in the formula I, . According to the method, based on specific catalysis of an alpha-amino acid dehydrogenase mutant obtained through directed evolution on thiomethyl alpha-butanone acid, efficient and high-stereoselectivity asymmetric reductive amination synthesis of corresponding thioamino acid is achieved.

The invention discloses an enzyme catalytic synthesis method of a dorzolamide chiral intermediate, which comprises the following steps: by taking methyl acetoacetate as a substrate, adding a buffer solution, the substrate, alcohol dehydrogenase, formate dehydrogenase, ammonium formate and coenzyme into a reaction container, reacting at 20-40 DEG C for 2-24 hours, extracting, separating and carrying out rotary evaporation to obtain the dorzolamide chiral intermediate (R)-3-methyl hydroxybutyrate, the alcohol dehydrogenase is a mutant derived from ADH1 of Ralstonia, and the amino acid sequence of the ADH1 is shown as SEQ ID No. 1. The method has the advantages of mild reaction conditions, high stereoselectivity, greenness, environmental protection, high conversion rate, high chiral purity, less enzyme dosage, tolerance to high-concentration substrates, low preparation cost and suitability for industrial production, avoids chiral resolution and heavy metal residues in products, and makes up for the defects of chemical methods.

The invention discloses an actinobacillus succinogenes gene engineering strain for highly producing succinic acid as well as a construction method and application of the actinobacillus succinogenes gene engineering strain. According to the actinobacillus succinogenes, rate-limiting enzymes, namely phosphoenolpyruvate carboxylase PPC and formate dehydrogenase FdoG, in a succinic acid synthesis route are overexpressed in series in the actinobacillus succinogenes, formic acid accumulation is reduced to provide more CO2 and NADH, and phosphoenolpyruvate is reinforced to fix CO2 to be converted into succinic acid, so that the yield of succinic acid is increased, and meanwhile, the yield of byproduct formic acid is reduced.

The invention discloses a method for synthesizing raspberry ketone through chemical enzyme stage combination and application, and belongs to the technical field of organic synthesis and biological engineering. The raspberry ketone is obtained through whole-cell catalysis by taking recombinant engineering bacteria of co-expressed olefine aldehyde reductase and formate dehydrogenase genes as a biocatalyst. When the adding amount of a genetic engineering bacterium E.coli BL21 (DE3) pRSF-AtQOR (I56S / A69H / A118N)-LbFDH is 20 g / L, the substrate HBA is 100 g / L, the sodium formate is 125 g / L, the NADP < + > is 0.5 mM, the reaction temperature is 37 DEG C, the pH is 7.5, the reaction time is 12 h, and the rotating speed is 220 rpm, the raspberry ketone concentration can reach 82.23 g / L. The problems of serious environmental pollution of a chemical method and low yield of a biological method are solved.

The modified ferroferric oxide comprises a magnetic Fe3O4 nano core, a magnetic nano core and a magnetic nano core, the Fe3O4 nano core is coated with the polydopamine photothermal conversion layer; a polyglutamic acid hydrophilic brush layer; the Ni ions are subjected to coordination modification on the polyglutamic acid hydrophilic brush layer; the preparation method of the modified ferroferric oxide comprises the following steps: 1) preparing Fe3O4 (at) PDA; (2) preparing Fe3O4 (at) PDA-PGA; (3) taking the Fe3O4 (at) PDA-PGA, and adding a Tris-HCl buffer solution, NaCl and nickel chloride hexahydrate, so as to obtain a Fe3O4 (at) PDA-PGA-Ni composite material; the invention relates to a method for measuring CO2 capture efficiency of immobilized formate dehydrogenase under illumination. The method comprises the following steps: a, preparing an FDH crude enzyme solution; b, preparing a Fe3O4 (at) PDA-PGA-Ni compound of the immobilized FDH; c, adding the Fe3O4 (at) PDA-PGA-Ni compound of the immobilized FDH into a PBS (Phosphate Buffer Solution), incubating at 36 DEG C for 10 minutes, adjusting the visible light intensity to enable the temperature of the Fe3O4 (at) PDA-PGA-Ni compound of the immobilized FDH to be constantly increased to 45 DEG C, adding NADH (Nicotinamide Adenine Dinucleotide), and introducing mixed gas of CO2 and N2; the concentration of CO2 at an outlet is measured, and the CO2 capture efficiency is calculated; measuring the concentration of formic acid in the reaction liquid, and calculating the CO2 conversion efficiency; the method has the characteristics that the carbon conversion cost is low, and the CO2 conversion efficiency is remarkably improved.

The application belongs to the field of green manufacturing, and discloses a preparation method of a pyridazine compound, comprising the following steps: (1) adding an enzyme catalyst, a bio-based furan and a co-substrate into a buffer solution to react, and removing cells after the reaction; (2) adding hydrogen peroxide or an oxidase and a titanium-silicon molecular sieve to react, and removing the titanium-silicon molecular sieve after the reaction; (3) slowly adding hydrazine under stirring at 0-60 DEG C to obtain the pyridazine compound after the reaction; the enzyme catalyst is at least one of the following: an alcohol dehydrogenase / glucose dehydrogenase double enzyme system, an alcohol dehydrogenase / formate dehydrogenase double enzyme system and a recombinant bacterium co-expressing one or both of the above two double enzyme systems. The application has the advantages of high selectivity, mild reaction conditions, environmental friendliness, fast reaction speed and no need for intermediate separation and purification.

The invention relates to the technical field of chemical preparation, and particularly discloses an ursodesoxycholic acid production process based on a double-enzyme method, which comprises the following steps: S1, carrying out first reaction on a chenodeoxycholic acid CDCA substrate and immobilized 7alpha-HSDH, NADP < + > and formate dehydrogenase FDH coenzyme circulation system in a continuous flow microchannel reactor under the conditions of 25 DEG C and pH 8.5 at a certain flow velocity to obtain a 7-oxy intermediate; s2, obtaining an ursodesoxycholic acid crude product; s3, obtaining a permeable liquid; s4, obtaining clear liquid; s5, obtaining an ursodesoxycholic acid finished product of which the purity is greater than or equal to 99 The technical scheme that the CDCA is directly converted to generate the UDCA by combining the 7alpha-HSDH and the 7beta-HSDH through two enzymatic reactions is effective and feasible, the industrial technical problem that the UDCA needs to be synthesized through a two-step enzymatic method at present is solved, a foundation is laid for biosynthesis of the UDCA, and the domestic blank is filled.

The invention belongs to the technical field of bioelectrocatalytic reduction of CO2, and relates to a charcoal-based enzyme electrode for CO2 reduction as well as a preparation method and application of the charcoal-based enzyme electrode. Firstly, SBA-15 is used as a template, microcrystalline cellulose is used as a carbon source, ordered porous biochar is constructed through temperature programming carbonization and template removal with strong base, then copper nanoparticles are formed in a CuSO4 electrolyte through cyclic voltammetry electrodeposition, and formate dehydrogenase and carbonic anhydrase are synergistically fixed on the surface of an electrode. The porous biochar provides a mass transfer channel with a high specific surface area and high efficiency, the copper nanoparticles improve the electron transfer efficiency, and formate dehydrogenase and carbonic anhydrase are combined to simulate a microbial carbon concentration mechanism, so that the CO2 concentration and reduction specificity of an interface are improved. Under the voltage of-0.3 V to-0.8 V (vs Ag / AgCl), CO2 is selectively reduced into formic acid, the process conditions are mild, the cost is low, and a feasible strategy is provided for high-selectivity formic acid preparation and CO2 recycling.

The invention relates to the technical field of catalysts, and discloses a photo-enzyme coupling CO2 conversion catalyst as well as a preparation method and application thereof. The catalyst comprises a composite carrier composed of a graphite phase carbon nitride nanosheet and MIL-125-NH2 (Ti), an amino silane coupling agent is grafted on the composite carrier; formate dehydrogenase is loaded on the composite carrier, and part or all of the formate dehydrogenase is covalently linked to the amino silane coupling agent. The catalyst disclosed by the invention has better light absorption capacity, CO2 adsorption capacity and mass transfer capacity, the adverse effect of the immobilization process on enzyme activity is smaller, and higher light-enzyme coupling CO2 conversion efficiency can be realized; in addition, the catalyst provided by the invention has a good protection effect on enzyme activity, and can effectively improve the stability and reusability of the enzyme.