1120 results about "Enzyme catalysis" patented technology

The invention discloses silk fibroin composite gel and a preparation method thereof, and relates to the technical field of tissue filling materials. The preparation method of the silk fibroin composite gel comprises the following steps: preparing silk fibroin and carrying out amination modification on the silk fibroin to obtain aminated silk fibroin; carrying out dynamic cross-linking on the aminated silk fibroin and the oxidized sodium alginate through a Schiff base bond by adopting an enzyme catalysis cross-linking process to obtain a dynamic cross-linked network solution; adding a temperature-sensitive component into the dynamic cross-linked network solution, and uniformly mixing to obtain a temperature-sensitive-dynamic dual-network pre-gel solution with temperature responsiveness; loading the active component into the temperature-sensitive-dynamic dual-network pregel solution to obtain a composite gel solution; incubating the composite gel solution to obtain primary gel; after disulfide bonds are introduced into the primary gel, cross-linking strengthening and sterilization are carried out, and the silk fibroin composite gel is obtained. Through temperature-sensitive-dynamic dual-network design, active components and disulfide bonds are introduced to strengthen crosslinking, and the problems that traditional injectable gel is irreversible, single in function, prone to collapse after being filled and the like are systematically solved.

The invention discloses a high-performance carbonyl reductase mutant and application thereof in synthesis of a series of chiral alcohols. The high-performance carbonyl reductase mutant is obtained by performing the following mutations on an amino acid sequence as shown in SEQ ID NO.2: the 69th alanine mutates into any one of lysine, leucine, aspartic acid or asparagine, and the 69th alanine mutates into any one of lysine, leucine, aspartic acid or asparagine; and / or the 100th glycine is mutated into any one of glutamic acid, serine, lysine or asparagine. Compared with a wild type enzyme, the thermal stability and the catalytic activity of the obtained mutant are remarkably improved, particularly, single point mutants Mut-A69N, Mut-G100E and Mut-G100N and a combined mutant Mut-A69N-G100N show excellent relative enzyme activity, thermal stability and substrate tolerance. The mutant has high catalytic activity and stability to various chiral alcohols, and the contradiction that the traditional carbonyl reductase is easy to inactivate at high temperature and the catalytic efficiency is difficult to achieve at the same time is solved. The invention provides an efficient and stable enzyme catalysis tool for green biological preparation of chiral drug intermediates.

The invention relates to a silk fibroin biological material with high beta-fold content and a preparation method thereof, and belongs to the technical field of biological extraction. The method comprises the following steps: preparing a regenerated silk fibroin solution; adding an enzyme catalytic promoter and an enzyme into the regenerated silk fibroin solution for incubation; the enzyme catalysis auxiliary agent is selected from one or more of a metal ion catalyst, a zwitterionic surfactant, a functionalized ionic liquid and a calcium phosphate nano-cluster; and centrifuging, cleaning and drying the obtained product to obtain the silk fibroin biological material with high beta-fold content. The silk fibroin material has higher beta-folding amount through primary structure modification, the space structure is more crystallized, and the amino acid sequence is more regular and ordered. The silk fibroin material provided by the invention has excellent mechanical properties, efficiency and yield, and is a biological protein material with great development potential.

The invention belongs to the technical field of separation of products for synthesizing antibiotics by an enzymic method, and particularly relates to a method for separating and recycling an amoxicillin product synthesized by a one-step method based on catalysis of kluyveromyces citricacidophilus beta subunit G385 mutant penicillin acylase. An immobilized penicillin acylase mutant is used for catalyzing penicillin potassium to synthesize amoxicillin in one step, a set of method for separating amoxicillin and phenylacetic acid is developed for an obtained reaction mixture, and the technical scheme mainly comprises the following steps: firstly, separating the immobilized penicillin acylase mutant from a reaction solution through filtration; then, amoxicillin is separated out through crystallization; and then crystallizing, separating and recovering phenylacetic acid through toluene extraction, back extraction and the like. According to the separation method, the amoxicillin can be rapidly and efficiently separated, the yield of the amoxicillin is relatively high, and the average crystallization rate reaches 93.22%; meanwhile, the separation and recovery effects of phenylacetic acid are good; and the extraction agent toluene can be recycled.

The invention provides a chiral resolution method and application of a 3-aryl substituted glutaric acid monoester compound, and the method comprises the following steps: taking a substance as shown in a formula I as a starting raw material, and carrying out alcoholysis to obtain a racemic intermediate as shown in a formula IV; and splitting the intermediate shown in the formula IV, and then dissociating to obtain the compound shown in the formula III, which has a single spatial configuration and an ee value of more than 99%. A cyclic anhydride substrate with a symmetrical structure is used and reacts with alcohol to obtain racemic 3-aryl substituted glutaric acid monoester, a target configuration with high chiral purity is obtained by screening some resolution reagents and solvents and performing salifying resolution on the 3-aryl substituted glutaric acid monoester, the operation is simple, the resolution reagents are easily available in the market and can be recycled, and the method is suitable for industrial production. The method has the advantages that an expensive chiral catalyst or a harsh enzyme catalysis condition is avoided, the reaction cost is reduced, amplified production is facilitated, the ee value of a target configuration can be increased to 99% or above, and the method is an economic and environment-friendly technical route which is simple in post-treatment and convenient for amplified production.

The present disclosure describes a Pseudococcus preparation and a preparation method and application thereof, the Pseudococcus preparation comprises an active ingredient, the active ingredient comprises a specific pro-inflammatory regression mediator (SPMs) and a pro-inflammatory regression mediator (PRMs), the specific proinflammatory regression medium and the proinflammatory regression medium are generated by catalyzing unsaturated fatty acid synthesized by micrococcus by own enzyme. According to the invention, the invention can provide the micrococcus preparation containing the specific proinflammatory regression medium and the proinflammatory regression medium as well as the preparation method and the application of the micrococcus preparation.

The present invention discloses an environmental monitoring system and a monitoring method, which relates to the field of environmental monitoring technology. The system accurately collects the abundance vector MAV of the microbial community within a fixed period, performs concentration and separation treatment, then adds a substrate reaction and monitors the enzyme catalytic reaction in the water sample in real time, generates an enzyme activity feature set EAF, combines the aerobic bacteria change rate Raer, the anaerobic bacteria change rate Ranaer and the correction factor F, forms a multidimensional data set Eac, and establishes a comprehensive water quality assessment algorithm based on this, thereby obtaining a comprehensive water quality detection and assessment index WQI, and realizing a quantitative assessment of the health status of water quality. By comparing with the water quality assessment standard threshold Sthe, an early warning notification is automatically triggered, water quality abnormalities are discovered and responded to in a timely manner, solving the problem that the traditional method cannot capture water quality changes in a timely manner, especially in terms of rapid response to environmental pollution and ecological changes. It has significant advantages.

The invention discloses a molecular modification design method for improving the catalytic activity and the thermal stability of citrobacter vinegaticus-derived hyaluronidase and expression application of the citrobacter vinegaticus-derived hyaluronidase. The interaction of citrobacter vinegaticus hyaluronidase active pocket amino acid residues and hyaluronate tetrasaccharide molecules is visually explored through Pymol, conservative analysis of pocket amino acid evolution is carried out through Consurf, and rational design is carried out through policies of FoldX for calculating enzyme-substrate binding free energy, so that the hyaluronidase catalytic activity is improved. A mutant with significantly improved enzyme activity is obtained through screening, and the thermal stability of the enzyme is further improved through PROSS design and an analysis strategy of mutation site amino acid evolution conservative property and position. According to the invention, the sequence, structure and function of hyaluronidase are deeply studied, and hyaluronidase is mutated from two aspects of stabilizing a catalytic structure and promoting a catalytic reaction, so that a high-activity enzyme with higher application value in industrial production is obtained.

The invention belongs to the technical field of engineered probiotics, and particularly relates to engineered probiotics for radiosensitization and tumor metabolism regulation as well as a preparation method and application of the engineered probiotics. The engineering probiotics comprise probiotics and core-shell type metal nanoparticles which are loaded on the probiotics and are synthesized through a biological directional mineralization effect; the core of the core-shell type metal nanoparticle is a palladium element, and the outer layer of the core-shell type metal nanoparticle is a palladium element and a gold element. The engineering probiotics provided by the invention have excellent enzyme catalysis performance and can target and retain in tumor sites, and through the synergistic effect of all components in the engineering probiotics, adenosine metabolism is effectively inhibited, secretion of pro-inflammatory cytokines is enhanced, tumor microenvironment is promoted to be converted into an inflammatory state, up-regulation of immune checkpoints is inhibited, and the immune checkpoints are inhibited. And T cells are inhibited from being transformed into depletion phenotype and transformed into effect type from depletion type. After the SBRT and the immune checkpoint inhibitor are combined for use, the growth of tumors can be effectively inhibited, and the effect of enhancing immune radiotherapy is achieved.

The invention discloses an endoglucanase truncated mutant and an application of the endoglucanase truncated mutant. According to the invention, based on endoglucanase BsEGL from bacillus subtilis DLG, a truncated mutant is constructed by deleting a carbohydrate binding domain (CBM) of the endoglucanase BsEGL, and the truncated mutant is named as BsEGL [delta] CBM. Compared with BsEGL, the truncated mutant BsEGL delta CBM has the advantages that the thermal stability, the pH stability, the organic solvent tolerance and the like are remarkably improved, and the activity of degrading phosphoric acid expanded cellulose and corncob xylan is obtained. In addition, the enzymatic activity of the truncated mutant BsEGL delta CBM is remarkably improved by 6.41%, and the catalytic efficiency is remarkably improved. Due to the characteristics, the mutant can be suitable for industrial processes such as feed processing, papermaking, biological energy sources, spinning and food brewing, and is particularly suitable for enzyme catalytic reaction under extreme environments such as high temperature and low pH. The invention provides an enzyme preparation tool with excellent performance for efficient degradation of cellulose biomass, and has important application value.

The invention provides an L-threonine aldolase mutant and application of the L-threonine aldolase mutant in catalytic synthesis of p-methylsulfonylphenylserine, the L-threonine aldolase mutant is derived from pseudomonas, threonine and p-methylsulfonylbenzaldehyde can be used as substrates, and the L-threonine aldolase mutant can be synthesized by using a one-step method. According to the present invention, L-threonine aldolase is adopted as a raw material, pyridoxal phosphate and reduced nicotinamide adenine dinucleotide are adopted as coenzymes, the synthesis of florfenicol intermediate product p-methylsulfonyl phenyl serine is obtained through catalysis, and compared with the wild type L-threonine aldolase, the mutant enzyme has high activity, can effectively improve the conversion efficiency, and can shorten the conversion time; the enzyme and the technical method provided by the invention have the advantages of mild reaction conditions and less pollution, and have good industrial prospects in the production of florfenicol synthesis intermediates.

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 composite gel for osteochondral repair as well as a preparation method and application of the composite gel. Main raw materials of the composite gel are regenerated silk fibroin (RSF) and recombinant humanized III-type collagen (rhCol-III), and auxiliary raw materials of the composite gel are sodium hyaluronate (HA), fibrinogen and thrombin. The preparation method comprises the following steps: firstly, mixing an RSF solution with an rhCol-III solution, obtaining RSF-rhCol particles through a freeze drying technology and mechanical pulverization, dispersing the particles in an HA solution to obtain composite gel, mixing the gel with a thrombin solution, catalyzing fibrinogen to be split and converted into fibrin by thrombin, and performing freeze drying to obtain the rhCol-RhCol-RhCol-RhCol-RhCol-RhCol-RhCol. And crosslinking the fibrous protein monomer to obtain the injectable composite gel preparation capable of immediately gelling. The prepared RSF-rhCol composite gel can adapt to a defect area, has a certain adhesion effect, has excellent biocompatibility, promotes migration of bone marrow mesenchymal stem cells, induces cell differentiation and promotes cartilage tissue regeneration.

The invention provides a method for preparing high-purity phosphatidylserine by a batch enzyme catalysis method based on dynamic choline removal, and belongs to the technical field of enzymatic reaction. According to the method disclosed by the invention, in a reaction system for preparing phosphatidylserine, a byproduct choline is selectively adsorbed by adopting resin. Choline in a reaction system is removed in real time through the integrated adsorption separation module, the enzyme inhibition problem is solved, and meanwhile, the PC conversion rate is larger than or equal to 98% and the PS purity is larger than or equal to 96% in combination with the immobilized enzyme circulation technology; the enzyme can be reused for more than or equal to 20 times (activity retention is more than or equal to 90%); and the choline residual concentration is less than or equal to 0.5 mM (an inhibition threshold value or less).

The invention provides a nitrilase mutant and application of the nitrilase mutant in synthesis of p-cyanobenzoic acid. The invention provides a nitrilase mutant capable of improving the activity of catalytic synthesis of p-cyanobenzoic acid, the mutant protein is a non-natural protein, and the mutant protein can significantly improve the activity of catalyzing p-phthalonitrile to generate p-cyanobenzoic acid. And the mutant protein mutates in two or more core amino acids related to enzyme catalytic activity of the wild nitrilase. The nitrilase mutant provided by the invention can significantly improve the yield of a nitrilase catalytic synthesis product.

The invention discloses a cathepsin B activated fluorescent probe which comprises a dicyanoisosulfone fluorescent report unit and a cathepsin B specific restriction enzyme cutting site GFLG peptide chain. When CTSB specifically cuts a GFLG peptide chain, a DCO molecule with an amino group with relatively strong electron donating ability is generated, so that the fluorescence emission behavior of the probe is rapidly activated through a one-step enzyme catalysis reaction. The invention also provides a preparation method of the cathepsin B activated fluorescent probe. The invention also provides a nano particle for packaging the cathepsin B activated fluorescent probe, and the nano particle is simple in preparation process and mild in reaction condition. The cathepsin B activated fluorescent probe is prepared into nanoparticles, and the nanoparticles can realize accumulation of the material in tumor tissues and accurate imaging in apoptotic tumor cells by enhancing the permeation retention effect, have better biocompatibility and can be used for detecting the apoptotic tumor cells.

The application discloses a galactosyltransferase mutant and application thereof in preparation of flavonoid compounds, and belongs to the technical field of enzymology. The application provides a UDP-galactosyltransferase mutant, which is obtained by site-directed mutation of one amino acid in the amino acid sequence of wild-type galactosyltransferase VcUFGT into alanine (A). The effects of a series of mutants H82A, V139A, G141A, P186A, N245A, V282A and S307A are verified in the examples, the mutants can perform enzyme catalysis reaction with quercetin and UDP-galactoside as substrates, and generate flavonoid compound hyperoside, and the specific enzyme activity of the mutants is more than 1.5 times that of the wild type, thereby providing an effective application basis for biosynthesis of flavonoid compounds.

The invention provides a carbonyl reductase mutant and application thereof in preparation of chiral alcohol compounds, and belongs to the technical field of gene engineering and enzyme catalysis. According to the invention, on the basis of the wild carbonyl reductase, the mutant with activity obviously superior to that of the wild carbonyl reductase is obtained through specific mutation, the selectivity and stability of the enzyme are improved, and the mutant has a wide application prospect in catalytic synthesis of chiral alcohol compounds.

The present application relates to the technical field of biocatalysis and biosynthesis, and specifically discloses a blue pigment and a biosynthesis method thereof. In the present application, an indigoidine synthetase and a 4′-phosphopantetheinyl transferase are expressed by a metabolically engineered strain to catalyze the biosynthesis of the blue pigment N-acetyl-indigoidine from glutamine and N-acetylglutamine, and a molecular structure of the blue pigment is inferred by mass spectrometry, nuclear magnetic resonance spectroscopy, etc. The present application achieves the catalytic synthesis of N-acetyl-indigoidine from glutamine and N-acetylglutamine in Escherichia coli (E. coli), Corynebacterium glutamicum (C. glutamicum), Saccharomyces cerevisiae (S. cerevisiae), and Streptomyces. Compared with indigoidine, N-acetyl-indigoidine has a maximum absorption wavelength of 584 nm, and a stable color having high brightness that is not easy to fade. Thus, the blue pigment shows an extensive application range and a promising industrial production prospect.

The invention discloses a dental plaque display type anti-caries invisible appliance based on synergism of iron-based nano-enzyme catalysis and traditional Chinese medicine caries inhibition, and belongs to the field of oral medical instruments. The preparation method of the appliance diaphragm comprises the following steps: preparing a composite material with the functions of catalytically degrading a bacterial biofilm matrix barrier and killing internal bacteria from polyethylene glycol terephthalate-1, 4-cyclohexanedimethanol (PETG) and a Fe3O4 nano-enzyme filler in a solution blending manner; and after the composite material is subjected to thermoplastic molding, a layer of molecularly imprinted polymer (MIP)-spiropyrane and chitosan-magnolol are coated with the fluorescent display agent which is prepared by mixing the molecularly imprinted polymer (MIP)-spiropyrane and the chitosan-magnolol. The iron-based nano-enzyme can catalytically degrade a bacterial biofilm matrix barrier and kill internal bacteria, and the MIP-spiropyrane fluorescent display agent can specifically display the accumulation condition of cariogenic bacterial plaque in the oral cavity under ultraviolet irradiation. The invisible orthodontic appliance can reduce or prevent caries while having the same orthodontic effect as other orthodontic appliances in the market.

The invention discloses an adenosine kinase mutant with high thermal stability and application thereof, and relates to the technical field of enzyme engineering, and the adenosine kinase mutant with high thermal stability is obtained by performing point mutation on the 253rd site or the 82nd site of adenosine kinase as shown in SEQ ID NO: 1. According to the invention, an adenosine kinase mutant is used as a biocatalyst in an enzyme catalysis manner; the method is used for enzymatic synthesis of 5 '-adenylic acid, adenosine and adenosine triphosphate react to prepare adenylic acid, generation of by-products and impurities in the reaction is reduced to a certain extent, production efficiency and purity of adenylic acid are improved, reaction conditions are mild in preparation in an enzyme catalysis mode, pollution to the environment is avoided, and the method is suitable for industrial production. And the high-thermal-stability adenosine kinase mutant is applied to adenylic acid preparation, so that the preparation efficiency and yield of adenylic acid can be improved.

The invention provides a saccharomyces cerevisiae engineering strain HYB05 capable of releasing astaxanthin in a controllable self-splitting manner. The saccharomyces cerevisiae engineering strain HYB05 is constructed by taking a basic strain for producing astaxanthin as a starting strain, introducing a coding sequence comprising cell wall lyase Lyc, a coding sequence of a signal peptide mutant F8A and a recombinant nucleic acid construct for expressing a regulatory element, and knocking out an SSD1 gene to relieve a translation inhibition effect. The cell wall lyase Lyc is found and disclosed by the inventor for the first time, and the amino acid sequence is shown as SEQ ID NO: 6; the amino acid sequence of the signal peptide mutant F8A is as shown in SEQ ID NO: 8. According to the engineering strain HYB05, by introducing the recombinant nucleic acid construct, the activation of the catalytic activity of cell wall lyase under specific conditions is realized, so that the cell wall of the saccharomyces cerevisiae is broken, and the astaxanthin synthesized by the saccharomyces cerevisiae is released. Compared with the prior art, the astaxanthin is produced through fermentation of the strain, the extraction steps are simplified, extraction of the astaxanthin in the yeast is achieved to the maximum extent, the yield of the astaxanthin is remarkably increased, and the technical effect is remarkable.

The invention relates to the technical field of mineral materials and nano-enzyme catalysis, in particular to an iron-containing layered silicate mineral nano-enzyme as well as a preparation method and application thereof. The preparation method comprises the following steps: carrying out ball milling on the iron-containing layered silicate mineral, carrying out centrifugal washing, and drying to obtain the iron-containing layered silicate mineral nano-enzyme. According to the method, silicate minerals with a layered structure and rich iron content are adopted as raw materials, the ball-milled layered silicate mineral nano-enzyme is successfully prepared through a ball milling method, the crystallinity of the minerals is reduced, the surface acidity of the minerals is improved, and low-spin iron is generated, so that the affinity of the ball-milled minerals to H2O2 is enhanced; h2O2 is catalyzed to generate singlet oxygen and superoxide free radicals, so that the peroxidase-like activity of the ball-milled minerals is improved. The enzyme-like activity is improved by 10 times, high catalytic activity can still be kept under the neutral condition, and the method can be used for colorimetric sensing and antibacterial application of small biological molecules such as glucose, hydrogen peroxide and glutathione.

The invention discloses a 4-hydroxyphenylacetic acid 3-hydroxylase mutant and application thereof, recombinant expression plasmids and corresponding genetically engineered bacteria M214V, R290N and E294I are constructed by using genes for coding the mutants (EcHpaBM214V, EcHpaBR290N and EcHpaBE294I) and nucleotide sequences for coding EcHpaC genes respectively, and after the genetically engineered bacteria disclosed by the invention are subjected to shake flask fermentation in a 30mL M9Y culture medium containing 50mu g / mL chloramphenicol for 24 hours, the yield of the mutants is increased, and the yield of the mutants is increased. The yield of the tanshinol is 748.04 mg / L, 895.55 mg / L and 793.36 mg / L. The method breaks through the limitation that the substrate generated by the 4-hydroxyphenylacetic acid 3-hydroxylase through catalysis is poor in specificity and low in catalysis efficiency, and the yield of the tanshinol is high. The method has the advantages of being high in catalytic efficiency, simple in process and environmentally friendly.

The invention discloses chimeric xylanase XynZT-1-Z5 and a construction method thereof, and relates to the technical field of molecular biology, and the technical key points are as follows: comparing the sequence of xylanase XynZT-1 with the sequence of xylanase from different thermoduric bacteria, analyzing the amino acid conserved site of the xylanase XynZT-1, and replacing the sequence beta5-beta6 of the thermoduric bacteria onto the XynZT-1 to obtain the chimeric xylanase XynZT-1-Z5. The chimeric enzyme XynZT-1-Z5 has efficient overexpression, good solubility and relatively strong temperature and pH stability, is particularly suitable for industrial enzyme catalytic reaction under the conditions of high temperature and extreme pH, and can be widely applied to the food industry, the wine brewing industry, the papermaking industry and the feed industry.

The invention provides a plasmid, an alpha-ketoglutaric acid engineering strain and a construction method and application of the alpha-ketoglutaric acid engineering strain, the strain is obtained by adopting a directional transformation method, transcriptional levels of katE, katG, sodB and grxA genes are up-regulated, a catR gene and a katA gene are heterologously expressed, a high-copy plasmid pETKG is carried, and the plasmid simultaneously expresses L-glutamate oxidase LGOX genes from Streptomyces sp and Chain A; the strain self-synthesized catalase eliminates H2O2 generated in the process of producing alpha-ketoglutaric acid through enzyme catalysis, engineering bacteria of L-glutamate oxidase LGOX expressed by pETKG plasmids are used for performing whole-cell catalysis on L-sodium glutamate to synthesize alpha-ketoglutaric acid, and the engineering strain is a high-level engineering strain capable of industrially and stably producing alpha-ketoglutaric acid.

The invention discloses a method for synthesizing ceramide NP through two-step enzyme catalysis, which comprises the following steps of: firstly, adding lipase into vegetable oil to catalyze and combine fatty acid to decompose into free fatty acid, and centrifuging to remove the lipase; a small amount of phytosphingosine is added into the hydrolyzed vegetable oil, the raw material vegetable oil is used as a solvent or a squalane system solvent for incubation, and then immobilized lipase is added for catalytic synthesis of ceramide. The invention establishes a two-enzyme method for efficiently synthesizing the ceramide NP compound, and the process is natural, environment-friendly and non-toxic. The reaction conditions are optimized through response surface analysis, meanwhile, the immobilized lipase shows excellent cycle performance, and the method is easy to operate, environmentally friendly, green and economical and has wide application prospects in the fields of cosmetics, health care products, biological medicine and the like.

The invention discloses a method for preparing inositol, particularly relates to a method for converting glucose into inositol through in-vitro multi-enzyme catalysis, and belongs to the field of enzyme catalysis preparation of inositol. According to the invention, the phosphoglucose synthase capable of realizing phosphorylation of glucose by using inorganic phosphate is screened, and the inositol can be generated through conversion by taking glucose as a substrate, so that a novel method for synthesizing the inositol through enzyme catalytic conversion is provided. In addition, the invention further provides hexokinase capable of improving the yield of inositol, and the conversion efficiency of glucose is greatly improved. Furthermore, the invention provides a multi-enzyme molecular machine containing the glucose phosphate synthase and / or hexokinase, the multi-enzyme molecular machine is applied to preparation of inositol, and the reaction steps are simple, green and pollution-free.

The invention discloses an azodicarbonamide ratio fluorescence detection method based on a CoOOH nanosheet loaded Au nano-cluster material, and belongs to the technical field of fluorescence sensing. The CoOOH nanosheet loaded Au nano-cluster material provided by the invention is prepared by carrying out a one-step electrostatic assembly method on AuNCs and a CoOOH nanosheet. The CoOOH nanosheet loaded Au nano-cluster material has the fluorescence property of the AuNCs and the biomimetic catalytic performance of the CoOOH nanosheet at the same time. The invention further provides an azodicarbonamide ratio fluorescence detection method based on the CoOOH nanosheet loaded Au nano-cluster material, and compared with an existing detection method, the azodicarbonamide ratio fluorescence detection method has the remarkable advantages that the method is high in selectivity; by utilizing a ratio fluorescence mechanism, interference caused by environment and instrument fluctuation can be effectively reduced, reliable quantitative analysis of azodicarbonamide is realized, and the method has the advantages of high sensitivity and high credibility and has a wide prospect in the field of fluorescence sensing detection.

The invention discloses an enzyme catalytic synthesis method of a luliconazole chiral intermediate, which comprises the following steps: adding a recombinant escherichia coli wet cell for expressing alcohol dehydrogenase, a recombinant escherichia coli wet cell for expressing formate dehydrogenase, a buffer solution, a substrate, a cosolvent, ammonium formate and a coenzyme into a reaction container, reacting at 25-35 DEG C for 2-24 hours, extracting, separating, and carrying out rotary evaporation, thereby obtaining the luliconazole chiral intermediate. The luliconazole chiral intermediate is obtained; the substrate is 2, 2 ', 4'-trichloroacetophenone; the alcohol dehydrogenase is a mutant of AKR3. The method is mild in reaction condition, environmentally friendly, high in regioselectivity and stereoselectivity, high in conversion rate, high in chiral purity, small in enzyme dosage, low in preparation cost and suitable for industrial production.