Abstract: Provided are a catalyst loaded with a TEMPO-type compound, and a preparation method and use thereof. The catalyst loaded with the TEMPO-type compound uses a polymer resin as a carrier, and the polymer resin is a polystyrene resin containing a chloromethyl or bromomethyl, herein the content of the chloromethyl or bromomethyl in the polymer resin is 0.5-5.0 mmol g?1.

Abstract: The disclosure provides a new crystal form of ertapenem sodium and preparation method therefor. The new crystal form of ertapenem sodium has 27 principal characteristic peaks in an X-ray powder diffraction diagram. The crystal form of ertapenem sodium of the disclosure has a rod-like crystal habit, which has a large particle size, is not prone to aggregation and is easier to dry, and a product with high crystallinity and high purity may be obtained by simple drying treatment. Meanwhile, the crystal form of the disclosure has better stability, and the crystal form may remain unchanged to a maximum extent in subsequent washing and drying processes, thereby further improving the purity of the product.

Abstract: The present invention provides a method for synthesizing a fluorine-containing chiral amine compound. The method comprised: reacting an amino donor with a fluorine-containing dihydroxy ketal compound under a catalysis of a transaminase to generate the fluorine-containing chiral amine compound, wherein the transaminase is derived from a plurality of strains. The transaminase of the present application has substrate specificity on the fluorine-containing dihydroxy ketal compound, and may effectively catalyze this type of the substrate to be converted into the fluorine-containing chiral amine compound. In addition, the transaminase has catalytic activity on a plurality of the fluorine-containing dihydroxy ketal compounds, and is relatively high in reaction selectivity and activity. The method catalyzed by the bio-enzyme is not only short in route, but also high in product yield, and the production using the method reduces cost, organic solvents and three wastes.

Abstract: The preparation method for a polypeptide includes the steps of constructing an engineering strain for fusion-expressing a polypeptide gene with a Sumo tag, and inducing the engineering strain for the soluble expression of a polypeptide, obtaining a crude protein containing a polypeptide precursor from the engineering strain by purification, cleaving the crude protein containing the polypeptide precursor by a Ulp1 protease to remove the Sumo tag, and purifying a cleavage product of the Ulp1 protease by a method of acetonitrile combined with heating precipitation or a method of precipitation with hexafluoro isopropanol to obtain the polypeptide.

Abstract: Provided are a cytochrome P450 enzyme mutant and an application thereof. The enzyme activity and anti-Markov oxidation selectivity of P450 derived from a wild-type strain of Bacillus megaterium undergo protein modification by means of directed evolution, thus improving enzyme activity and selectivity, and developing a series of P450 enzyme mutants that may be used for industrial production.

Abstract: Described herein are a PVA membrane immobilized enzyme and a preparation method therefor. The PVA membrane immobilized enzyme includes a PVA porous membrane and an enzyme entrapped on the PVA porous membrane. The PVA porous membrane is a three-dimensional structured PVA porous membrane. The enzyme is any one selected from transaminase, D-lactate dehydrogenase, cyclohexanone monooxygenase, ketoreductase, alkene reductase, nitrilase, ammonia lyase, amino acid dehydrogenase, imine reductase, alcohol dehydrogenase, ammonium formate dehydrogenase, glucose 1-dehydrogenase and mutants thereof. The three-dimensional structured PVA porous membrane is used as a carrier to immobilize an enzyme in an entrapment manner. After entrapping and immobilizing the enzyme in the PVA porous membrane, the enzyme is stable, and cannot be easily leached out in the process of use. The PVA porous membrane is suitable for use in continuous flow biochemical catalysis, and has wide applicability to enzymes.

Abstract: Provided is an immobilized enzyme, a preparation method and use thereof. The immobilized enzyme includes an enzyme and an amino resin carrier for immobilizing the enzyme, and the enzyme is selected from any one of the following enzymes: transaminase, ketoreductase, monooxygenase, ammonia-lyase, ene reductase, imine reductase, amino acid dehydrogenase, and nitrilase. The amino resin carrier is an amino resin carrier modified by a cross-linking agent, and the cross-linking agent is a cross-linking agent treated by a polymer. By means of modifying the amino resin carrier with the cross-linking agent treated by the polymer, the enzyme immobilized on the amino resin carrier may easily form a network cross-linking, such that the immobilization effect of the enzyme is more stable, thereby the recycling efficiency of the enzyme is improved.

Abstract: Provided is a ketoreductase mutant and applications thereof. Compared with an amino acid sequence shown in SEQ ID NO: 1, the amino acid sequence of the ketoreductase mutant includes at least one of the following mutation sites: L198P, S96W/I/L/V, M194L or L1526, herein “/” means “or.” The ketoreductase mutant provides high selectivity and high activity with respect to a substrate ketone, and the application of the mutant in a catalytic reduction reaction of a ketone substrate may increase the production efficiency of a chiral alcohol compound corresponding thereto.

Abstract: Provided are a transaminase mutant and an application thereof. Compared with an amino acid sequence shown in SEQ ID NO:1, an amino acid sequence of the transaminase mutant includes at least one of the following mutation sites: L166, K149, K146, A168, H73, F133, H82, E24, V194, T294, A295, G235 and F236. The mutant of the present invention has the improved catalytic activity for a transammonization reaction of ketone substrates, and is suitable for industrial production of chiral amines.

Abstract: Provided are a transaminase mutant and use thereof. The transaminase mutant has an amino acid sequence obtained by mutation of an amino acid sequence shown in SEQ ID NO:1, the mutation at least includes one of the following mutation site combinations: T7C+S47C, Q78C+A330C, V137C+G313C, A217C+Y252C and L295C+C328C; or the transaminase mutant has an amino acid sequence which has the mutation sites in the mutated amino acid sequence and has 80% or more identity with the mutated amino acid sequence. The transaminase mutant realizes the change of protein structure and functions, reduces the enzyme amount, increases the enantiomeric excess (ee) value of a product, and reduces the difficulty of post-processing, so that the transaminase mutant may be suitable for industrial production.

Abstract: Provided is a method for synthesizing a chiral diamine compound. The synthesizing method includes: converting a substrate represented by Formula I into a chiral diamine compound in Formula I by using a transaminase, herein n=1˜10, an R group represents an alkyl, a cycloalkyl, a heteroatom-containing alkyl, a heteroatom-containing cycloalkyl, a heteroatom-containing aryl, an amide compound residue, or an ether compound residue, and a hetero atom is at least one from among O, S and N; R1 and R2 are the same or not the same, the R1 and R2 are respectively and independently hydrogen, a C1-C3 alkyl, or an amino protecting group; and the transaminases are derived from a plurality of strains.

Abstract: Disclosed are a modified epoxy resin immobilized enzyme, a preparation method therefor and an application thereof. Herein, the preparation method includes the following steps: modifying an epoxy resin, adding a polyethyleneimine to a modified epoxy resin for further modification, and then adding an enzyme to be immobilized and a glutaraldehyde for immobilization, to obtain the modified epoxy resin immobilized enzyme. The epoxy resin is modified, the polyethyleneimine is added to the modified epoxy resin for the further modification, and an aldehyde group in the resin and an amino group in the polyethyleneimine are covalently bound to each enzyme, then it is activated by the bifunctional reagent glutaraldehyde.

Abstract: Described herein are an immobilized enzyme, and a preparation method therefor and a use thereof. The immobilized enzyme includes activated PEI and an enzyme covalently bonded to the activated PEI, where the enzyme is selected from any one of a transaminase, a ketoreductase, a monooxygenase, an ammonia lyase, an ene-reductase, an imine reductase, an amino acid dehydrogenase and a nitrilase.

Abstract: Provided are a co-immobilized enzyme, a preparation method and use thereof. The co-immobilized enzyme includes: an amino resin carrier, a main enzyme, and a coenzyme. The main enzyme and the coenzyme are co-immobilized on the amino resin carrier, herein the main enzyme is covalent-immobilized on the amino resin carrier, and the coenzyme is immobilized on the amino resin carrier by a mode of covalent and/or non-covalent; and the main enzyme is selected from any one of the following enzymes: transaminase, amino acid dehydrogenase, imine reductase, ketoreductase, enoyl reductase, and monooxygenase. The main enzyme and the coenzyme thereof are co-immobilized on the amino resin carrier for co-immobilization, so the activity and the recycling efficiency of the enzyme are improved.

Abstract: Disclosed are a ketoreductase mutant and a method for producing a chiral alcohol. The ketoreductase mutant has an amino acid sequence obtained by the mutation of the amino acid sequence shown in SEQ ID NO: 1, and the mutation includes a mutation siteK200H. In the present disclosure, the mutant obtained by mutation takes a ketone compound as a raw material, the chiral alcohol may be efficiently produced by stereoselective reduction, and the stability is greatly improved, which is suitable for popularization and application to the industrial production of the chiral alcohol.

Abstract: An amino acid sequence of the transaminase mutant is an amino acid sequence obtained by a mutation of an amino acid sequence is shown in SEQ ID NO: 1. The mutation occurred at least one of the following mutation sites: G17V, L36P, Q40H, G69Y, H70T, L73A, V77G, V77S, V77T, A78I, Y130M, Y130V, Y130T, N132I, N132T, K141S, K142S, K142T, R143P, G144F, G144W, G144Y, E145D, E145S, E145G, K146R, L148A, L148I and the like.

Abstract: The present disclosure discloses a continuous preparation method for a penem intermediate MAP. The continuous preparation method includes the following steps: step S1, in a column-type continuous reactor, using a rhodium-loaded catalyst to catalyze 4-nitrobenzyl(R)-2-diazo-4-((2R,3S)-3-((R)-1-hydroxyethyl)-4-oxoazetidin-2-yl)-3-oxopentanoate to generate a cyclization reaction so as to form a first intermediate, herein the rhodium-loaded catalyst is loaded in the column-type continuous reactor, and the rhodium-loaded catalyst has the following structural formula: step S2, performing an esterification reaction on the first intermediate, a diphenyl chlorophosphate and a diisopropylethylamine in a second continuous reactor, to obtain a product system containing the penem intermediate MAP; and step S3, performing crystallization treatment on the product system, to obtain the penem intermediate MAP.

Abstract: Provided are a transaminase mutant and a method for producing a chiral amine using the same. An amino acid sequence of the transaminase mutant is an amino acid sequence obtained by mutation occurred in an amino acid sequence as shown in SEQ ID NO: 1, and the mutation includes at least one of the following mutation sites: position 3, position 5, position 8, position 25, position 32, position 45, position 56, position 59, position 60, position 84, position 86, position 164, position 176, position 178, position 180, position 187, position 197, position 206, position 207, position 242, position 245, position 319 and position 324.

Abstract: An amino acid sequence of the monooxygenase mutant is obtained by mutation of an amino acid sequence shown in SEQ ID NO: 1, and the mutation at least includes one of the following mutation sites: 45-th site, 95-th site, 106-th site, 108-th site, 114-th site, 186-th site, 190-th site, 191-th site, 249-th site, 257-th site, 393-th site, 436-th site, 499-th site, 500-th site, 501-th site, 503-th site, 504-th site, 559-th site, and 560-th site.

Abstract: Provided are a proline hydroxylase and uses thereof. The proline hydroxylase comprises having the amino acid sequence of SEQ ID NO: 2 with the exception of a mutation of one or more amino acids; wherein the mutation of one or more amino acids must comprises E27K, and the mutation of one or more amino acids selected from the group consisting of: H14R, L16N, T25R, F26L, E27K, D30S, S33N, E34N, E34G, E34L, E34S, E34D, Y35W, Y35K, S37W, S37F, S37E, S37N, S37T, S37C, W40F, K41E, D54G, H55Q, S57L, I58T, I58Y, I58A, I58R, I58V, I58S, I58C, K86P, T91A, F95Y, C97Y, I98V, K106V, K106T, K106Q, F111S, K112E, K112R, S154A, K162E, L166M, I118F, I118V, I118R, H119R, H119F, I120V, K123D, K123N, K123Q, K123S, K123I, K123T, T130N, D134G, V135K, N165H, D173G, K209R, I223V and S225A, and having proline hydroxylase activity.