484 results about "Biocatalysis" patented technology

The invention relates to the technical field of biological catalysis, and discloses an L379A mutant enzyme for preparing rebaudioside I. The enzyme is obtained by the following mutations generated by UGT76G1: leucine of the 379th amino acid sequence is mutated into alanine; the enzyme can be applied to RA in-vitro conversion preparation of RI with higher utilization value, the conversion rate is higher than 50%, and the enzyme activity is remarkably improved by 7 times compared with the original enzyme catalytic conversion enzyme activity. The UGT76G1 mutant disclosed by the invention has the advantages that (1) the blank is filled, and a special enzyme catalyst capable of realizing efficient in-vitro synthesis of rebaudioside I (RI) is provided for the first time; 2) efficiency jump: the catalytic conversion rate is greatly increased from original about 7% to more than 50% (increase gt; and 3) stability and reliability: in the cross-scale reaction of 10mL to 5L, the catalyst has stable catalytic performance, shows excellent industrial application potential, and is suitable for popularization and application.

The invention relates to the technical field of biological catalysis, and discloses an M88V mutant enzyme for preparing rebaudioside I. The M88V mutant enzyme is obtained by the following mutations generated by UGT76G1: M88V: methionine of the 88th amino acid sequence of UGT76G1 is mutated into valine. Aiming at the technical bottlenecks of low RI yield and poor selectivity in the existing RA biotransformation process, an efficient solution is provided for the UGT76G1 mutant developed by the invention. The mutant can specifically catalyze conversion from RA to RI, and the conversion rate of a target product is remarkably increased to 65% or above from general less than 7%. The excellent catalytic performance of the catalyst is verified in milliliter-level to upgraded reaction systems, the key problems of conversion efficiency and process amplification are solved, and a foundation is laid for large-scale green manufacturing of a high-added-value product RI.

The invention relates to the technical field of biological catalysis, and discloses an M88A mutant enzyme for preparing rebaudioside I. The enzyme is obtained by the following mutations generated by UGT76G1: M88A: methionine of the 88th amino acid sequence of UGT76G1 is mutated into alanine; the key conversion rate of RI is greatly increased from 7% to 85% (increased by more than 12 times), and the utilization rate and economical efficiency of raw materials are remarkably improved; the method can be perfectly reproduced under any test scale (10mL, 100mL and 5L), the conversion rate is always anchored to be more than 85%, and incomparable stability and amplification potential are shown.

The invention relates to the technical field of biological catalysis, and discloses an L204A mutant enzyme for preparing rebaudioside I. The enzyme is obtained by the following mutations generated by UGT76G1: L204A: leucine of the 204 amino acid sequence of UGT76G1 mutates into alanine; the high-efficiency in-vitro enzymatic preparation of the rebaudioside I (RI) is realized for the first time, the catalytic efficiency of the rebaudioside I (RI) is improved by more than 7 times (from about 7% to more than or equal to 50%) compared with that of an original enzyme, the high-efficiency performance can be stably maintained under different scales of 10mL to 5L, a solid foundation is laid for industrial application of the RI, and the preparation method is simple, controllable in process condition and suitable for popularization and application.

The invention relates to the technical field of enzyme engineering, in particular to an acylase mutant and a biosynthesis method of (R)-2-aminobutanol.The acylase mutant comprises at least one of mutation of the following sites relative to wild type acylase with the amino acid sequence being SEQ ID NO: 1: T106, A625 and S413. The acylase mutant disclosed by the invention is used for synthesizing (R)-2-aminobutanol through biological catalysis, and the reaction route is as follows: the catalytic efficiency of a product is remarkably improved by the mutant, and the ee value of a chiral product is improved; according to the synthesis method, the problem of chiral resolution in the prior art is solved, the production cost is reduced, and pollution is reduced.

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 a preparation process of a moxidectin pouring solution. The preparation process comprises the following steps: S1, respectively preparing a moxidectin initial solution and a thickener premixed solution; s2, the moxidectin initial solution and the thickener premixed solution are stirred and mixed uniformly at the temperature of 25-35 DEG C to form a mixture A, the stirring speed is 150-250 rpm, and the mixing time is 10-20 min; s3, adding auxiliary materials into the mixture A, and uniformly mixing to form a mixture B; s4, supplementing the second solvent into the mixture B to the total amount, uniformly mixing, and filtering to obtain a moxidectin pouring solution; the method comprises the following steps: dissolving a moxidectin raw material in a first solvent to obtain a moxidectin initial solution, and synthesizing the moxidectin raw material by using biological catalysis; the first solvent is a nano-emulsion formed after homogenization. According to the preparation method, the solubility of the medicament, the stability of the preparation and the transdermal drug delivery efficiency are improved by improving the solvent system and controlling the process in the preparation process of the moxidectin pouring solution.

The invention discloses a synthesis method of 2 '-fluoro-2'-deoxyguanosine. According to the method, thymine nucleoside phosphorylase and purine nucleoside phosphorylase are used as biocatalysts, and 2 '-fluoro-2'-deoxyuridine and guanine are used as substrates to synthesize the 2 '-fluoro-2'-deoxyguanosine. Compared with a chemical method, the production cost is greatly reduced; meanwhile, the synthesis method can effectively solve the problem that the product is difficult to separate.

The invention discloses a method for converting rutin into quercetin through biological catalysis, and belongs to the technical field of biological catalysis. The method comprises the following steps: mixing rutin and a solvent to obtain a quasi-solid rutin suspension; mixing the paraphlonia plant endogenous enzyme and the quasi-solid rutin suspension for enzymolysis, and then filtering, washing and drying to obtain quercetin; the solvent is methanol with the concentration of 5-50%, ethanol with the concentration of 5-50% or acetone with the concentration of 10-50%. The method is mild in biological catalytic reaction condition, does not generate pollutants such as acid waste liquid, and meets the environmental protection requirement. Efficient conversion of quercetin is realized by adopting a quasi-solid rutin strategy (rutin suspension reaction system), and the problem of substrate solubility limitation is solved. High-purity quercetin can be obtained through simple washing, the product purity is larger than 95%, and food and clinical application can be met.

The invention relates to the technical field of biological catalysis, and discloses an M88F mutant enzyme for preparing rebaudioside I. The enzyme is obtained by the following mutations generated by UGT76G1: M88F: methionine of the 88th amino acid sequence of UGT76G1 is mutated into phenylalanine; and the substrate rebaudioside A (RA) can be efficiently and directionally converted into rebaudioside I (RI) with higher value. In an optimized reaction system, the conversion rate stably reaches 40%, and the catalytic efficiency is improved by more than 5-8 times compared with the common reference enzyme. The enzyme preparation has the characteristics of high catalytic activity, mild reaction conditions, simplicity and convenience in operation and the like, shows good stability and reproducibility in laboratory research and large-scale production, and has a wide industrial application prospect.

The invention provides a recombinant O-succinyl-L-homoserine mercaptotransferase mutant, a coding gene thereof, a recombinant vector containing the coding gene of the mutant, a recombinant genetically engineered bacterium obtained by converting the recombinant vector, and application of the recombinant O-succinyl-L-homoserine mercaptotransferase mutant in preparation of L-methionine. The novel recombinant O-succinyl-L-homoserine mercaptotransferase mutant with high activity and high selectivity is used as a catalyst to be applied to catalysis of O-succinyl-L-homoserine for synthesis and preparation of L-methionine, a catalytic reaction system is optimized, the feed ratio and reaction conditions in the reaction are finely regulated and controlled, and the yield of L-methionine is increased. The recombinant O-succinyl-L-homoserine mercaptotransferase mutant has high enzyme activity under the optimal reaction condition, the catalytic efficiency is further improved, the purposes of reducing cost and improving efficiency are achieved, and the method has important significance in promoting industrial application of biological catalysis preparation of L-methionine.

The invention relates to the technical field of polypeptide biosynthesis, in particular to a method for biosynthesizing acetyl tetrapeptide-5, a composition containing the acetyl tetrapeptide-5 and application of the composition. According to the method, low-cost natural amino acid is used as a substrate, specific aminopeptidase or a mutant thereof is used as a biocatalyst, amino acid ligase and polyphosphatase are combined, and acetyl tetrapeptide-5 can be efficiently synthesized through a one-step enzymatic reaction. Compared with a traditional chemical synthesis method, the method has the advantages of low raw material cost, no use of an organic solvent, mild reaction conditions, high regioselectivity, environmental friendliness, low production cost and the like. The invention also provides a composition containing acetyl tetrapeptide-5 and dipeptide-15, the inhibition effect of the composition on grease secretion is obviously better than that of a single component, and the composition has a synergistic oil control effect under a specific ratio.

The invention discloses a salicylic acid decarboxylase mutant and application of the salicylic acid decarboxylase mutant in degradation of salicylic acid. The mutant is obtained by carrying out site-specific modification on salicylic acid decarboxylase NahG, and specifically, valine at the 46th site, tyrosine at the 380th site and leucine at the 382nd site of an amino acid sequence shown in SEQ ID NO.2 are mutated into cysteine, phenylalanine and phenylalanine at the same time. The mutant shows remarkably improved catalytic efficiency, and salicylic acid can be efficiently and thoroughly decarboxylated and converted into catechol under mild conditions. The invention also provides a coding gene, a recombinant vector and an engineering bacterium of the mutant. The mutant not only can be used for efficient bioremediation of salicylic acid pollution, but also can effectively break through and accelerate a naphthalene catabolism pathway due to the characteristic of directionally generating catechol, and has important application value in the fields of environmental pollution abatement and biological catalysis.

The invention belongs to the field of photovoltaic technology, and provides a photovoltaic packaging adhesive film, a preparation method and application thereof, and a recovery method of a photovoltaic module. The photovoltaic packaging adhesive film comprises the following raw materials in specific amounts: matrix resin, a low-temperature cross-linking agent, a low-temperature catalyst, a silane coupling agent, an ultraviolet light stabilizer, an organic peroxide initiator, an assistant cross-linking agent and an alcohol compound serving as a reversible dynamic transesterification reaction group, the matrix resin is EVA (ethylene vinyl acetate), POE (polyolefin elastomer) and / or ethylene-butylene copolymer; the low-temperature cross-linking agent is a carboxylic ester type cross-linking agent, a bisoxazoline type cross-linking agent and / or a metal-organic ligand cross-linking agent with the reaction temperature of 150 DEG C or below; the low-temperature catalyst is a polynuclear metal catalyst, a solid acid resin catalyst, a solid base catalyst and / or a bimetallic MOF derivative catalyst with the reaction temperature of 150 DEG C or below. The adhesive film can realize recycling of the adhesive film and a photovoltaic module in a more efficient and energy-saving manner, is wide in adaptability, and has high crosslinking degree, peel strength and PID resistance.

This invention discloses a highly efficient asymmetric synthesis method for aryl propionic acid drug molecules. The method uses racemic aldehydes as substrates and thiamine pyrophosphate-dependent enzymes as biocatalysts. Electrolysis is performed in a diaphragm-free electrolytic cell under conditions of an external redox dielectric and constant current, in a buffer salt solution and nitrogen atmosphere. This method achieves dynamic kinetic oxidation of aldehydes via bioelectrocatalysis, enabling the high-efficiency and high enantioselectivity synthesis of aryl propionic acid drug molecules. The method eliminates the need for redox agents and additives, is simple to operate, operates under mild reaction conditions, and is green and sustainable. This invention applies bioelectrocatalysis to asymmetric synthesis, providing a green and efficient new enzymatic synthetic route for the asymmetric synthesis of aryl propionic acid drug molecules, which has significant application prospects in the pharmaceutical industry.

The present invention provides methods for the manufacture of amides from their corresponding nitriles using a biocatalyst and purification of the manufactured amides using membrane filtration. The invention also relates to methods of using membrane filtration for isolating and reusing biocatalysts. More specifically, the inventive method produces a crude aqueous acrylamide (AMD) by feeding acrylonitrile (AN), water, and a biocatalyst with nitrile hydratase activity into a reactor. After complete conversion of AN to AMD, the crude reaction is directly purified by membrane filtration to produce a purified AMD monomer (without nitrile hydratase activity) and a recovered biocatalyst with nitrile hydratase activity. Purified monomer (e.g., AMD without enzyme activity) in the permeate may be stored, transported, and used for polymerization. Biocatalyst in the retentate retains enzymatic activity and may be reused.

The invention relates to the field of biological catalysis, and discloses a method and a device for generating ursodesoxycholic acid through continuous flow catalysis of chenodeoxycholic acid. Comprising the following steps: (1) in the presence of first immobilized cells, carrying out a first catalytic reaction on a reaction system containing chenodeoxycholic acid to obtain a first product, the first immobilized cells expressing 7 alpha-hydroxysteroid dehydrogenase and lactic dehydrogenase; the concentration of the chenodeoxycholic acid in the reaction system is 40 to 70 mM; (2) adjusting the pH value of the first product to 8-9, and then in the presence of a second immobilized cell, carrying out a second catalytic reaction on the first product after the pH value is adjusted to obtain ursodesoxycholic acid, and the second immobilized cell expresses 7 beta-hydroxysteroid dehydrogenase and glucose dehydrogenase. According to the method, the stable activity of a catalytic system is kept, efficient conversion of high-concentration CDCA is achieved, meanwhile, precipitation of 7-KLCA is thoroughly avoided, and the synthesis efficiency, the product purity and the process continuity of UDCA are remarkably improved.

The application discloses a method for synthesizing high-concentration furfuryl alcohol by using a double-enzyme coupling method, and the method comprises the following steps: mixing alcohol dehydrogenase freeze-dried bacterial powder and glucose dehydrogenase freeze-dried bacterial powder to serve as a catalyst, using furfural as a substrate, using glucose as a co-substrate, using NADP + or NAD + as a coenzyme, using a buffer solution with a pH value of 4-9 as a reaction medium to form a reaction system, and reacting completely under the condition of 20-50 DEG C and 400 rpm, then separating and purifying the reaction liquid to obtain furfuryl alcohol. The biological catalytic system is suitable for furfural with a concentration higher than 500 mM, and 100% conversion of 500 mM can be realized within 10 hours. The substrate continuous flow process is further used to eliminate the inhibition of aldehyde substrates on the biological catalyst, the concentration of the product can reach 1500 mM, and the highest space-time yield can reach 235 g / (L.d).

The invention discloses a lactic dehydrogenase LdLDH mutant and application thereof, a wild type LdLDH enzyme derived from Lactobacillus delbrueckii subsp. Bulgaricus is taken as a research object, the amino acid sequence of the wild type LdLDH enzyme is designed and modified to obtain a mutant protein, the mutant protein is named as LdLDH-M, the amino acid sequence of the mutant protein is shown as SEQ ID NO.2, the Tm value of the LdLDH-M is 77.1 DEG C, the Tm value of the LdLDH-M is 77.1 DEG C, and the Tm value of the LdLDH-M is 77.1 DEG C; compared with the wild type LdLDH enzyme, the Tm value of the LdLDH enzyme is increased by 27.4 DEG C, and the enzyme activity of the LdLDH enzyme is increased by 1.4 times. Therefore, compared with the wild type LdLDH enzyme, the LdLDH-M has remarkably improved thermal stability and enzymatic activity, and is more beneficial to widening the application range of the LdLDH-M in the fields of high-temperature biological research, industrial biological catalysis and the like.

The invention belongs to the technical field of biology and medicinal chemistry, and discloses a transaminase mutant, a method and application of the transaminase mutant in preparation of a key chiral intermediate of rotigotine, and the preparation method of the key chiral intermediate of rotigotine is a biological conversion method using the transaminase mutant as a main enzyme. The amino acid sequence of the transaminase mutant is an amino acid sequence obtained by mutating the amino acid sequence of wild type transaminase as shown in SEQ ID NO.1, the amino acid sequence of the transaminase mutant has a mutation site in the mutated amino acid sequence, and the amino acid sequence of the transaminase mutant has more than 90% of homology with the mutated amino acid sequence. When 5-methoxy-2-tetralone (a compound shown in a formula III) is catalyzed by an enzymatic reaction method to prepare a key chiral intermediate (S)-5-methoxy-1, 2, 3, 4-tetrahydronaphthalene-2-amine (a compound shown in a formula II) of rotigotine, the product is chiral purity gt, and the purity gt of the key chiral intermediate (S)-5-methoxy-1, 2, 3, 4-tetrahydronaphthalene-2-amine is gt. The yield of the biological catalysis step is gt; 95%.

The invention relates to a pyridoxal phosphate (PLP) synthesis method based on cascade biological catalysis and capable of realizing ATP autonomous regeneration. The method comprises the following steps: constructing engineering bacteria for co-expressing pyridoxal phosphate kinase (PdxK) and polyphosphate kinase (PPK) to prepare a whole-cell catalyst or a crude enzyme solution; under the condition that ATP does not need to be exogenously added, pyridoxal, ADP and polyphosphate serve as substrates, cascade reaction is carried out under the action of a catalyst, ATP is regenerated in situ, and PLP synthesis is driven. A wild enzyme system is adopted, and the yield of PLP exceeds 95% within 4 hours of reaction; after PdxK mutation optimization, the reaction time is shortened to about 2 hours, and conversion can be completed. According to the invention, a polyphosphate-driven ATP regeneration system is coupled with PLP biosynthesis, so that the cofactor cost is remarkably reduced, a feasible technical path is provided for industrial biological manufacturing of PLP, and the application of polyphosphate in cyclic regeneration of cofactors is expanded.

The invention relates to the technical field of biological catalysis engineering, in particular to a method for improving the solubility of stevioside Reb M and Reb D through enzyme catalysis modification, which comprises the following steps: dissolving Reb M and Reb D with the purity of more than or equal to 90% in a buffer solution with the pH value of 4.0-8.0, adding a non-starch glycosyl donor and composite glycosyl transferase according to the enzyme-substrate mass ratio of 0.05-0.5, and optimizing the enzyme addition sequence to realize specific site modification; reacting for 6-24 hours under the conditions that the pH is 5.5-6.0 and the temperature is 60-85 DEG C, and controlling the proportion of DP2-3 to be 60-80%; hPLC sampling is carried out every 4-6 hours, and the unreacted substrate is less than or equal to 20%; the solubility of the product is greater than or equal to 25g / L, and the sweet taste of the product is pure. According to the method, the solubility of Reb M / Reb D is improved, the structure of a directional glycosylation controlled product is realized, and pure sweetness is reserved; the reaction is mild and controllable, enzyme inactivation is simple and free of residues, industrialization is easy, and stevioside application upgrading is promoted.

The application relates to a biosynthesis method of gold nanoparticles, which comprises taking gold ions as gold nanoparticle precursors, taking microorganisms as biocatalysts to carry out a reduction reaction of the gold ions to obtain the gold nanoparticles; the reaction condition is mild, the method is environment-friendly, simple and easy to operate, and the synthesized gold nanoparticles have high biocompatibility and catalytic activity. The gold nanoparticles prepared by the method can be applied to the fermentation hydrogen production of hydrogen production bacteria.

The invention relates to the technical field of waste smoke utilization, in particular to a method for preparing 2, 5-dihydroxypyridine based on biotransformation of crude nicotine, and the method comprises the following steps: S1, obtaining a genetic engineering strain for accumulating 2, 5-dihydroxypyridine by enabling an hpo gene in pseudomonas putida S16 not to be expressed; s2, culturing the genetic engineering strain obtained in the step S1 in a culture medium to obtain a thallus suspension serving as a biocatalyst; s3, enabling the thallus suspension obtained in the step S2 to be in contact with a substrate containing nicotine, and carrying out biotransformation reaction to generate the 2, 5-dihydroxypyridine. The method provided by the invention has the advantages of simpler reaction system, fewer by-products and easiness in product separation and purification when being used for preparing the 2, 5-dihydroxypyridine.

The invention provides a yeast rich in polyphosphate and application of the yeast in synthesis of adenosine triphosphate, and the content of polyphosphate is 15-35 wt% based on the dry matter mass of the yeast rich in polyphosphate; according to the method disclosed by the invention, the yeast is subjected to phosphorus starvation culture and then is subjected to phosphorus supplementary culture, so that the yeast is enriched with polyphosphates, and the yeast rich in polyphosphates is obtained; then, the yeast rich in polyphosphate is used as a biocatalyst to synthesize ATP, that is, self-related enzyme systems, mainly including adenosine kinase, endo-polyphosphate kinase and excision-polyphosphate kinase, in the yeast rich in polyphosphate are used for replacing exogenous polyphosphate kinase and adenosine kinase, and in addition, ATP is synthesized by using the yeast rich in polyphosphate as a biocatalyst. According to the method for synthesizing ATP by using the yeast rich in polyphosphate as the biocatalyst, exogenous adenosine kinase and exogenous polyphosphate kinase do not need to be additionally added, the cost is low, the efficiency of converting adenosine into ATP is high, the batch-to-batch stability of the yeast is realized, and the enzyme activity of the self-related enzyme system is high.

The invention relates to the technical field of biological engineering and biological catalysis, in particular to two alcohol dehydrogenase mutants and application thereof. The invention discloses a pair of alcohol dehydrogenase mutants CpAR2-M3 (F85A / F192L / E209A) and CcPAR-M1 (V158L) with opposite stereoselectivity, wherein the corresponding amino acid sequences of the alcohol dehydrogenase mutants CpAR2-M3 (F85A / F192L / E209A) and CcPAR-M1 (V158L) are respectively shown as SEQ ID No.5 and SEQ ID No.6. The invention further discloses a preparation method of the alcohol dehydrogenase mutants. The method can be applied to asymmetric reduction of hemiacetal compounds, synthesis of corresponding key intermediate axial chiral biaryl benzyl alcohol compounds with complementary stereoselectivity is achieved, the highest yield can reach 95%, the highest stereoselectivity can reach 99% e.e., and the method is suitable for industrial production. A green, efficient and high-stereoselectivity synthesis strategy is provided for subsequent enantiocomplementary synthesis of the compounds.

The invention belongs to the field of biological catalytic materials, and particularly relates to a palladium monatomic-based biological catalyst, a biological catalytic compound thereof, a preparation method and application of the biological catalyst. The invention designs a coordination complex (Pd-Tpb) based on a palladium single atom from the beginning and finds that the coordination complex can be used as a biocatalyst with radiation-enhanced ROS (reactive oxygen species) generation activity and tumor killing performance for the first time. On the basis, Pd-Tpb is used for modifying lactobacillus probiotics, the obtained biocatalytic compound is used for biocatalysis and radiosensitization treatment, and it is found that the biocatalytic compound has the remarkably improved radiosensitization effect and has the effect of eradicating primary and metastatic tumors.

The present application relates to a kind of whole cell catalytic synthesis N-acetyl-trans-4-hydroxyproline method, belong to biological catalysis technical field.The present application is constructed by mutating acyltransferase and heterologous expression gene engineering bacteria, successfully realized using whole cell catalytic synthesis N-acetyl-trans-4-hydroxyproline, avoids the problem of enzyme instability, reduces the separation cost of enzyme and product.On this basis, the present application improves the yield of N-acetyl-trans-4-hydroxyproline by optimizing catalytic time, cell addition amount, the type and addition amount of additive, and the yield reaches 239mg / L.The present application is a kind of N-acetyl-trans-4-hydroxyproline preparation method with lower cost, lower equipment demand, more suitable for industrial production.

This invention belongs to the field of enzyme engineering technology, specifically relating to an enzyme engineering method and its application in biocatalysis. Based on the excellent performance optimization capabilities of the deep learning-guided protein sequence design model ProteinMPNN, this invention developed a computational design framework called DeepPCD. This framework integrates deep learning-guided sequence design, physical energy calculation, and molecular dynamics simulation, constructing a three-level intelligent screening process from "massive generation" to "high-precision prediction." Using this strategy, nitrile hydrolase (EC 3.5.5.1) was mutated, resulting in a nitrile hydrolase mutant with improved stability, specificity, and enzyme activity, capable of efficiently and specifically catalyzing the hydrolysis of nitrile compounds to carboxylic acids.

The invention provides a beta-glucosidase mutant from Paenibacillus phyllosphaerae, and a method for efficiently preparing rare ginsenoside F2 by using an enzyme method and a whole-cell method, and particularly provides a beta-glucosidase mutant from Paenibacillus phyllosphaerae and a method for efficiently preparing rare ginsenoside F2 by using a beta-glucosidase mutant from Paenibacillus phyllosphaerae. The method comprises the following steps: engineering-modifying beta-glucosidase by utilizing computer-aided design, constructing a mutant library, carrying out primary screening and secondary screening to obtain two high-activity mutants Q7D and G189A, constructing a combined mutant Q7D / G189A, transforming the combined mutant Q7D / G189A into genetically engineered bacterium escherichia coli for expression, and producing F2 by utilizing a whole-cell method. Finally, the highest yield of 18.01 g / L is achieved after the reaction is carried out for 10 h in a 5 L fermentation tank and is 6.82 times that of a shake flask, and the purity of F2 reaches up to 82.20% after the reaction is carried out for 24 h finally. The invention not only provides an efficient solution for large-scale production of rare saponin F2, but also provides a transferable framework for designing and engineering diversified glycoside hydrolase in customized biological catalysis.