75results about "Multi-enzyme systems" patented technology

The invention provides a preparation method and application of a bacterium-enzyme-magnetic nano-cluster composite detoxicating agent, the detoxicating agent takes a Fe3O4-HAP nano-cluster as a magnetic core, the surface of the detoxicating agent is subjected to functional modification, a composite flora composed of bacillus amyloliquefaciens, lactobacillus plantarum and saccharomyces cerevisiae is loaded to form sclerotia, a chitosan shell layer is constructed on the outermost layer, and the bacterium-enzyme-magnetic nano-cluster composite detoxicating agent is prepared. And a compound enzyme consisting of aflatoxin Bdegrading enzyme, zearalenone hydrolase and deoxynivalenol invertase is immobilized. The carrier can efficiently and synchronously degrade aflatoxin, vomitoxin and zearalenone, and can realize rapid separation and recovery through an external magnetic field.

The invention belongs to the technical field of soil organic matter improvers, and discloses a soil organic matter improver based on a compound microbial agent and a preparation method thereof, and the soil organic matter improver comprises the following components: a compound enzyme, a modified chitosan microsphere complex, a modified polyelectrolyte, an inhibitor L-methionine sulfoximine, a substrate, an auxiliary factor and the compound microbial agent. A compound enzyme system is used as a core, soil ammonium ions are circularly and efficiently assimilated through glutamine synthetase / glutamate synthetase, and the soil ammonium ions are converted into organic nitrogen; and energy guarantee is provided by virtue of polyphosphate kinase. The modified chitosan microsphere complex is used for protecting and slowly releasing enzyme and a substrate, and controlled release of the inhibitor L-methionine sulfoximine is realized through the modified polyelectrolyte, so that carbon-nitrogen balance of soil is regulated and controlled. The system has a synergistic effect with a compound microbial agent, the problem of soil organic matter reduction caused by excessive nitrogen application is effectively solved, meanwhile, the crop root biomass is remarkably increased, and the soil organic matter content is increased.

The invention discloses a functional microbial agent for purifying breeding sewage and a preparation method of the functional microbial agent, and belongs to the technical field of functional microbial materials. The microbial agent is composed of a core functional unit and a porous carrier according to a mass ratio of 3: 1 to 5: 1, the core functional unit comprises a symbiotic engineering flora and a compound enzyme system in a volume ratio of 7: 1, and the symbiotic engineering flora is compounded by sphingomonas aeruginosa, paenibacillus splendidus and acinetobacter hemolyticus according to a ratio of 1: 1: 1; the porous carrier is a low-cost porous carbon material prepared on the basis of an animal waste blood / iron complex. Through systematic cooperation of a specific functional flora, a compound enzyme system and a special porous carrier, synergism of adsorption protection and biodegradation is achieved, the stress resistance and purification efficiency of the microbial agent in an antibiotic and heavy metal combined pollution environment are remarkably improved, and the industrial problem that an aquaculture sewage biological treatment system is prone to collapse is particularly solved; meanwhile, waste is treated by waste, and the method is low in cost and suitable for large-scale application.

The invention discloses a Pickering IL emulsion microreactor constructed on the basis of temperature-sensitive double-enzyme Janus spheres and application of the Pickering IL emulsion microreactor to enhanced catalysis of conversion of CO2 into formic acid. A Janus emulsion template method is combined with a photocuring technology to synthesize a nano-scale dual-functional temperature-sensitive Janus sphere, one half of the nano-scale dual-functional temperature-sensitive Janus sphere is a hydrophilic hydrocarbon polymer, the other half of the nano-scale dual-functional temperature-sensitive Janus sphere is a hydrophobic fluorocarbon polymer, and rapid inversion of a Pickering emulsion can be realized by utilizing temperature sensitivity. The method comprises the following steps: preparing a Janus particle hydrophilic / hydrophobic hemisphere, selectively immobilizing formate dehydrogenase FDH and carbonic anhydrase CA on the Janus particle hydrophilic / hydrophobic hemisphere to obtain a temperature-sensitive FDH-Janus-CA composite sphere, introducing a fluorine-functionalized ionic liquid to regulate and control the internal microenvironment of a microreactor, and constructing a Pickering IL emulsion microreactor. The CO2 absorption capacity is increased by 47 times compared with that of a pure water system, the formic acid yield reaches 27.5 mM and is higher than the maximum value 22.8 mM of current enzyme electrocatalysis, the storage and catalysis stability is good, and an efficient and sustainable scheme is provided for CO2 recycling.

The invention provides a leavening agent and a preparation method thereof, and belongs to the technical field of leavening agents. According to the scheme, an efficient fermentation system is constructed through the synergistic effect of bacillus amyloliquefaciens, bacillus licheniformis, lactobacillus plantarum and saccharomyces cerevisiae, in the initial stage, the bacillus amyloliquefaciens secretes amylase and phytase degradation substrates, and conditions are created for the saccharomyces cerevisiae to metabolize monosaccharide to generate flavor substances and vitamins; in the middle stage, bacillus licheniformis decomposes protein to release amino acid, lactobacillus plantarum regulates pH through lactic acid metabolism to inhibit infectious microbes, and a multi-layer encapsulated compound enzyme preparation is introduced to synergistically degrade a complex substrate; in the later stage, fructooligosaccharide in the prebiotics-buffer stabilizer promotes proliferation of probiotics, mannitol and chitosan protect the activity of thalli and the function of metabolic enzymes, and the fermentation environment is stabilized; according to the whole system, the functionality, the environmental adaptability and the storage stability of the leavening agent are improved through flora synergistic metabolism, dynamic fermentation condition control and the action of the multifunctional additive, and the diversified requirements in industrial production are met.

The present invention relates to enzymatic reactor cells and related methods of use, e.g., to produce a compound or product by using an enzymatic reactor cell, wherein the enzymatic reactor cell includes a surface, a linker, and one or more enzymes.

The invention discloses a method for catalytically synthesizing adenosine triphosphate by using microcrystalline cellulose carrier purification immobilized enzyme, which comprises the following steps: preparing bacterial suspension by using escherichia coli wet thalli and Tris-HCL buffer solution, crushing, centrifuging, and taking supernate to obtain crude enzyme; adding a microcrystalline cellulose carrier and the wet thalli into the crude enzyme liquid, and washing the immobilized enzyme after immobilization to obtain the immobilized enzyme; the method comprises the following steps: adding immobilized enzyme adenosine kinase and immobilized polyphosphate kinase into a substrate system with the volume of 1L according to a weight ratio, and starting a catalytic reaction under the conditions that the pH is 5.5-7.5 and the temperature is 20-40 DEG C; after catalysis is finished, the immobilized enzyme adenosine kinase and the immobilized polyphosphate kinase are subjected to solid-liquid separation with reaction liquid through a Buchner funnel, the immobilized enzyme continues to be subjected to catalytic reaction, and the method for preparing the immobilized enzyme is simple in mode, low in cost, easy to industrially produce, high in stability and capable of being repeatedly used for many times, and products are easy to separate.

The application discloses a multi-enzyme immobilized enzyme for glucose-6-phosphate synthesis, which is obtained by immobilizing three enzymes, starch debranching enzyme mutant AcDBE_E131R, maltodextrin phosphorylase mutant AtMalP_N188D and glucose phosphomutase mutant TkPGM_F360P, on a resin. The application also discloses a preparation method and application of the multi-enzyme immobilized enzyme. The application screens enzyme mutants with good thermal stability and high catalytic activity by site-directed mutagenesis of starch debranching enzymes, maltodextrin phosphorylases and glucose phosphomutases from different sources, and prepares the multi-enzyme immobilized enzyme AcDBE_E131R+AtMalP_N188D+TkPGM_F360P. The multi-enzyme immobilized enzyme is used as a catalyst, maltodextrin is used as a substrate, and glucose-6-phosphate is efficiently synthesized by one-pot method, and the conversion rate reaches 71.2%. After 50 times of reuse, the relative enzyme activity of the multi-enzyme immobilized enzyme still remains 85.1%.

The invention provides fruit anthocyanin as well as an extraction method and application thereof. The extraction method comprises the following steps: S1, raw material pretreatment: selecting fresh fruits, removing impurities, cleaning, draining surface moisture, performing crushing treatment, then performing vacuum drying, and finally performing crushing to obtain fruit powder; s2, extracting three-layer microspheres: preparing an extraction substrate solution, adding the three-layer microspheres, and stirring for 8-12 minutes at the temperature of 40-45 DEG C and the rotating speed of 100-150r / min; the rotating speed is kept at 100-150 r / min, the power is set to be 300 W, the temperature is 42-45 DEG C, working is conducted for 4-6 min, pause is conducted for 1-2 min, and intermittent ultrasonic extraction is conducted for 3-4 h; after the extraction is finished, centrifuging an extracting solution, and collecting supernate to obtain a crude extracting solution; and S3, obtaining a final product of the fruit anthocyanin: filtering, adsorbing, eluting and purifying, concentrating and drying the crude extract to obtain the fruit anthocyanin. A targeted attachment-step conversion-enrichment protection and release linkage system is formed among the three-layer structures of the three-layer microspheres, and 1 + 1 + 1gt is realized; and 3, due to the synergistic effect, the activity and the quality stability of the anthocyanin are guaranteed while the extraction rate of the anthocyanin in the fruits is increased.

To provide an immobilized enzyme composition for hexose production or an improved process for hexose production. [Solution] The present invention relates to an immobilized enzyme composition for the preparation of hexoses. Examples of hexoses include tagatose, psicose, fructose, allose, mannose, galactose, altrose, talose, sorbose, gross, idose, and inositol. The present invention also relates to an enzymatic process for preparing hexoses from sugars by contacting starch derivatives with the immobilized enzyme composition of the present invention.

The invention discloses a bio-enzyme preparation for catalytic degradation of grease as well as a preparation method and application thereof, and belongs to the technical field of bio-enzymes. The gel carrier is formed by cross-linking a chitosan-lauric acid-dopamine conjugate and Fe < 3 + > ions through covalent coupling; the bonding adaptive compound enzyme is a mixed enzyme of which the surface is grafted with a catechol group through an N-hydroxysuccinimide-dopamine ester modifier; the technical bottleneck that efficient catalysis, firm fixation, convenient recovery and long-acting stability of an industrial cleaning enzyme preparation are difficult to consider at the same time is solved, and the prepared biological enzyme preparation has the high-activity recovery rate, excellent recycling stability, intelligent separation and recovery characteristics and efficient broad-spectrum cleaning capacity; and the method has remarkable industrial application advantages and market competitiveness.

The invention relates to a process for preparing TEMTP through turntable extraction, and belongs to the technical field of organic annular peroxides. According to the process, a rotating disc reactor is adopted to integrate a photocatalysis-enzyme catalysis-chemical catalysis triple synergistic system, and the problem of cross inactivation of a catalyst is solved through partition design and nano sieve membrane isolation. A peptide-guided magnetic MOF photocatalytic carrier is adopted as a core, a Fe3O4 magnetic core of the peptide-guided magnetic MOF photocatalytic carrier achieves magnetic field driving, a ZIF-8 frame loads a TiO2-Ag photocatalyst, PNIPAM temperature-sensitive hydrogel fixes halogenase, and the intelligent enzyme control characteristic is achieved. According to the method, TEMTP is taken as a raw material, a reaction is carried out under light source irradiation and magnetic field driving, extraction and purification are carried out through fluoroether HFE-7500 at normal pressure, a high-purity TEMTP product is finally obtained, and unification of reaction efficiency, selectivity and green production is achieved.

The invention relates to an enzyme-loaded porous gel capable of being quickly reconstructed, a preparation method and a flow microreactor, amphiphilic nanoparticles are used as a stabilizer, bubbles are introduced into a polymer solution and polymerization crosslinking is induced to form a uniform porous gel structure with an adjustable pore diameter, the amphiphilic nanoparticles can be adsorbed on a gas-liquid interface to stabilize the bubbles, and the porous gel structure can be quickly reconstructed. Enzyme molecules can be loaded on the surface to form a porous reaction medium with catalytic activity. The obtained porous gel has good mechanical strength, controllable aperture and porosity, and can realize rapid reconstruction of a porous structure, so that the application scene of a microreactor is widened, and the effective service cycle of enzyme is remarkably prolonged when the microreactor is blocked.

The invention relates to a Janus nano silicon dioxide particle with double-enzyme split-phase distribution as well as a preparation method and application of the Janus nano silicon dioxide particle. The preparation method comprises the following steps: by taking gamma-glycidyl ether oxypropyl trimethoxy silane and octyl trimethoxy silane as modifying agents, modifying nano SiO2, so as to prepare nano SiO2Janus particles with an epoxy group; then adding glucose oxidase to covalently bind with epoxy groups on the surfaces of the particles, and stirring for reaction to obtain silicon dioxide particles with glucose oxidase immobilized on the hydrophilic side; and finally, adding a CALB enzyme solution, stirring and reacting, and centrifuging, washing and drying after the reaction is completed, so as to obtain the Janus nano silicon dioxide particles with double-enzyme split-phase distribution. Epoxy groups are introduced and can react with amino groups to achieve enzyme immobilization, use of a cross-linking agent in the immobilization process is avoided, immobilization conditions are mild, and operation is easy.

This invention relates to a carrier-free immobilized enzyme and its preparation method, wherein the carrier-free immobilized enzyme is a catalytically active inclusion body of the enzyme crosslinked with glutaraldehyde, and the enzyme's end (especially the N-terminus) is tagged with histidine-leucine-histidine-leucine-histidine-leucine.

This invention belongs to the field of microbial inoculant technology and discloses a corn stalk decomposing agent product and its preparation method. The decomposing agent is composed of microbial inoculants, enzyme preparations, nutrient adjuvants, and a modified carrier in specific weight proportions. The microbial inoculants are a compound of Bacillus subtilis, Trichoderma viride, and Aspergillus niger; the enzyme preparation is a complex of cellulase, xylanase, and laccase; the nutrient adjuvant is a mixture of urea, sucrose, and potassium dihydrogen phosphate; and the carrier is an alkali-modified complex of wheat bran and straw powder. The preparation method includes preparing each microbial inoculant separately, alkali-modifying the carrier, and mixing all components. This invention also discloses the application method of this decomposing agent in corn stalk decomposition. This product achieves rapid, efficient, and thorough decomposition of corn stalks through the synergistic effect of multiple components, and the preparation process is stable, the product has a long shelf life, and it is suitable for large-scale production and field application.

The present invention specifically relates to an enzyme combination comprising a glucosidase enzyme, a secoisolariciresinol dehydrogenase enzyme and a glucose dehydrogenase enzyme, which are capable of carrying out reactions to convert secoisolariciresinol diglucoside (SDG) into matairesinol with a high yield. The invention also relates to the uses of the above combination, and to methods for producing matairesinol.

The invention relates to the field of bio-enzyme catalysis, and discloses a bio-enzyme catalysis process for reducing agent modification, which comprises the following steps: S1, enzyme solution preparation: mixing a bio-enzyme preparation with a buffer solution with the pH value of 5-8 to prepare an enzyme solution with the enzyme concentration of 0.5-5.0 g / L; the biological enzyme preparation is a compound enzyme composed of one or more of cellulase, hemicellulase and lignin degrading enzyme; s2, enzymolysis modification: mixing a carbonaceous reducing agent raw material with the enzyme solution prepared in the step S1 according to a mass ratio of 1: 5-1: 20, carrying out an enzymolysis reaction at 40-60 DEG C, and meanwhile, applying ultrasonic waves to carry out auxiliary treatment; the power of the ultrasonic waves is 100-500 W, and the frequency of the ultrasonic waves is 20-40 kHz And S3, solid-liquid separation: after the reaction is finished, separating the modified reducing agent from the enzymatic hydrolysate to obtain the solid modified reducing agent. Compared with the prior art, the method has the advantages that a biological enzyme catalysis process for modifying the reducing agent is provided, the energy consumption is low, the environment is protected, the cost is low, and the performance of the reducing agent can be remarkably improved.

The present invention relates to a nano-vesicle reactor and a method for producing the same. Specifically, the present invention provides a composition for producing a nano-vesicle reactor, characterized by comprising a first vesicle containing a first receptor on its surface and a second vesicle containing a second receptor on its surface, wherein the first receptor and the second receptor are bound with each other through a ligand system so that the first vesicle and the second vesicle are fused to form the nano-vesicle reactor ; and a method for producing the above nano-vesicle reactor. The nano-vesicle reactor of the present invention can be utilized in production of energy inside a cell through cellular enzymatic reaction, a drug delivered system, and a diagnostic system.

The invention discloses an organic selenium composition, a preparation method and application of the organic selenium composition in preparation of antitumor drugs, and belongs to the technical field of enzyme-containing preparations, the preparation method comprises five steps of carrier activation, immobilization of double enzymes, immobilization of a third enzyme, in-situ synthesis and separation and purification; aminated mesoporous silica nanoparticles are used as a carrier, selenocysteine beta-lyase, UDP-glucosyltransferase and glucose dehydrogenase are immobilized through glutaraldehyde crosslinking, an immobilized three-enzyme system is constructed, and organic selenium is synthesized in situ by catalyzing substrates such as jerusalem artichoke polysaccharide and selenocysteine in a nitrogen atmosphere. And separating and purifying to obtain the organic selenium composition. The problems of easy enzyme inactivation, large coenzyme dosage, poor product stability and the like in the prior art are solved, and the prepared organic selenium composition is high in selenium content, concentrated in molecular weight, excellent in bioavailability and low in toxicity, can be used as an active component for preparing antitumor drugs, and has a good application prospect.

A deproteinized natural rubber latex and preparation method thereof are provided. A combined immobilization technology of a multi-enzyme system is introduced to form a multi-enzyme synergistic effect, such that high-concentration enzymes contact with proteins in natural rubber latex. Meanwhile, due to the complexity and diversity of latex proteins, the catalytic properties of different enzymes are effectively combined to realize a multi-enzyme cascade reaction, and the proteins are hydrolyzed gradually and orderly. In addition, an immobilized enzyme filter screen is introduced, and the filter screen is lifted and circulated to make rubber particles collide with high-density enzyme carrier and then complete mixing evenly to reduce damages to the rubber particles when surface proteins are removed, thereby maintaining original physical properties of the latex. The rubber latex has high stability, reusability, is easy for separation and continuous operation, and is environmentally friendly.

The invention relates to the technical field of bioengineering, in particular to a preparation method and application of an immobilized enzyme. Compared with free enzyme, the immobilized enzyme prepared from the selected specific resin can be reused and is high in stability; the immobilized enzyme is easily separated from a reaction system, so that a product purification process can be simplified, and the product yield and quality are improved. Moreover, by immobilizing a sugar nucleotide producing enzyme and four sialyltransferases, a new strategy suitable for large-scale synthesis of functional sugar chains such as sialylated sugar chains is developed, the stability of related enzymes is improved, long-term storage and reutilization of the enzymes are realized, and the production cost of the functional sugar chains is reduced.

The application discloses a preparation method of oligomaltose and application thereof, and the method comprises the following steps: taking high straight corn kernels as raw materials, and carrying out microwave cooking and pulse electric field synergistic treatment, so that the corn kernels are softened, the starch-protein compound structure is destroyed, the corn kernels have more excellent processing performance, and more oligomaltose can be prepared by compounding alpha-amylase, alpha-glucoside transglycosidase and maltotriose enzyme; and the prepared immobilized composite enzyme has high thermal stability, can guarantee the efficiency and effect of enzymolysis reaction, has high controllability of enzymolysis, can be repeatedly used, reduces production cost, has high overall yield and purity, can replace serum protein to feed young animals, promotes the development of an immune system, increases the immunity of an organism, improves the disease resistance and growth, and reduces the probability of diseases.

The invention provides a method for synchronously treating kitchen waste and garden waste by using a compound enzyme, which specifically comprises the following steps: preparing a compound enzyme preparation, pretreating the kitchen waste and the garden waste, and performing enzymolysis on the compound enzyme preparation to prepare an organic fertilizer, and belongs to the technical field of solid waste treatment. According to the prepared compound enzyme preparation, the silicon-based polyphosphazene and zinc double-layer complex is adopted for immobilizing the compound enzyme, the original activity of the compound enzyme can be guaranteed, and the immobilization efficiency of the compound enzyme is effectively improved; the temperature-sensitive unit responds to release the unlinking agent in the enzymolysis temperature rising process, the aldehyde group contained in the unlinking agent interacts with the hydroxyl group in the welan gum double-helix network, the compound enzyme in the immobilized enzyme structure is released, and the enzyme activity is improved. Protein, fat, cellulose and the like in the kitchen waste and the garden waste are decomposed, conversion of organic solid waste is accelerated, the reaction process is 2-3 h, and rapid recycling of the waste is achieved.

The present application relates to the technical field of bioengineering, and particularly relates to a preparation method and application of immobilized enzyme. Compared with free enzyme, the immobilized enzyme prepared by selecting specific resin can be reused and has high stability; the immobilized enzyme is easy to separate from the reaction system, and can simplify the product purification process, improve the product yield and quality. Moreover, the present application develops a new strategy suitable for the large-scale synthesis of functional sugar chains such as sialylated sugar chains by immobilizing one kind of sugar nucleotide synthase and four kinds of sialyltransferase, improves the stability of related enzymes, realizes long-term preservation and reuse of the enzymes, and reduces the production cost of functional sugar chains.

A construction method of double immobilized recombinant yeast engineering bacteria, engineering bacteria, a catalyst and an application in synthesis of rebaudioside. In the present invention, yeast is used as the chassis cell, an ordered self-assembled multi-enzyme cascade surface display system is constructed, and carrier-free immobilization is coupled with uridine piphosphate glucose (UDPG) in-situ regeneration technology, resulting in a novel yeast whole cell catalyst with double immobilization of enzymes and bacteria. The catalyst uses cheap steviol glycoside (St) and rebaudioside A (Reb A) as substrates, and can synthesize rebaudioside (Reb D / M) in an efficient and economical one-pot method without additional addition of expensive UDPG. The synthesis process does not require cumbersome operations including cell disruption, enzyme separation, purification and immobilization, and avoids bottleneck problems in the related art including material transmembrane transport resistance, mass transfer resistance and product hydrolysis by intracellular enzyme. The catalyst has reusability, and can realize continuous and high-intensity rebaudioside biosynthesis through strain rejuvenation, which provides strong technical support for its industrial application.

The invention relates to the technical field of feed additives, in particular to a compound enzyme preparation for indoor aquaculture of marine economic animals. The compound enzyme preparation is prepared from the following components in parts by weight: 13 to 22 parts of protease, 8 to 15 parts of amylase, 12 to 26 parts of xylanase, 10 to 20 parts of phytase, 8 to 13 parts of beta-glucanase, 5 to 10 parts of cellulase, 5 to 10 parts of pectinase, 5 to 10 parts of chitinase and 3 to 5 parts of glucose oxidase. The preparation process comprises the following steps: step 1, single enzyme pretreatment; step 2, grouping and compounding; step 3, mixing and sieving; and 4, packaging a finished product. The phytase, the beta-glucanase, the chitinase and the glucose oxidase are added, so that the feed additive can adapt to different types of feeds, and meanwhile, through the synergistic effect of the phytase, the beta-glucanase, the chitinase and the glucose oxidase, the utilization rate of the feeds can be increased, the culture environment is optimized, and green ecological aquaculture is realized.

The present invention relates to a process to carry out two or more enzymatic reactions in a reaction medium, wherein the process comprises providing a system comprising at least two substantially homogeneous and colloidal populations of magnetic nanoparticles (MNPs) and applying one or more external alternating magnetic field to produce the simultaneous or sequential activation of the enzymes functionalized on the surface of each population of MNPs so that the enzymatic reactions can be thermally activated.

The invention discloses a method for preparing a pesticide intermediate through catalysis of multienzyme and coenzyme co-immobilized nanogel, and belongs to the field of synthesis of pesticide intermediates. Comprising the following steps: preparing a nano-emulsion from sodium alginate, halogenase, flavin reductase, glucose dehydrogenase, coenzyme flavin adenine dinucleotide, coenzyme nicotinamide adenine dinucleotide, water, an organic solvent and an emulsifier; performing cross-linking on the nano-emulsion and a calcium chloride solution to form multi-enzyme and coenzyme co-immobilized nano-gel; the preparation method comprises the following steps: preparing 1-methyl-3-ethyl-4-chloro-5-pyrazole carboxylic acid from ethyl 1-methyl-3-ethyl-5-pyrazole formate, nanogel, sodium chloride, glucose, sodium hydroxide, hydrochloric acid and water; and preparing the 1-methyl-3-ethyl-4-chloro-5-pyrazole carboxylic acid from the ethyl 1-methyl-3-ethyl-5-pyrazole formate. The multi-enzyme and coenzyme co-immobilized nanogel has the advantages of high catalytic efficiency, good selectivity, high repeated utilization rate, easy recovery and the like, and the production efficiency of the 4-chloro-3-ethyl-1-methylpyrazole-5-formic acid is effectively improved while dangerous reagents such as chlorine and sulfonyl chloride are replaced for a green chlorination process.

The invention relates to a biocatalyst for organic synthesis, comprising a controlled porosity silica (CPS) as support material, wherein the pore diameter is between about 20 and about 100 nm, said support material comprising an amino-functionalized surface; and one or more catalytically active enzyme(s) immobilized on the support material. The invention also relates to uses of such biocatalysts and methods of manufacture thereof.