11 results about "Dehalogenase" patented technology

The invention provides a method for improving the stability of dehalogenase through an arabinogalactan covalent cross-linking immobilization technology. The method comprises the steps of constructing and expressing a recombinant dehalogenase strain, purifying the recombinant dehalogenase and carrying out dehalogenase covalent cross-linking immobilization. The arabinogalactan is used for covalently crosslinking the dehalogenase, so that the thermal stability of the dehalogenase and the stability of the dehalogenase under different pH conditions and in an organic solvent can be remarkably improved.

This invention discloses a haloalkane dehalogenase HLD-1405, its encoding gene, and its applications, belonging to the field of biotechnology. The amino acid sequence of this haloalkane dehalogenase HLD-1405 is shown in SEQ ID NO.2, and its encoding gene is shown in SEQ ID NO.1. This haloalkane dehalogenase HLD-1405 exhibits high catalytic activity towards 1,3-dibromopropane, with optimal operating conditions of 40℃ / pH 9.0, and retains 100% activity in 20% methanol. This haloalkane dehalogenase HLD-1405 can efficiently degrade short-chain haloalkanes and also has the ability to degrade pollutants such as DDT and hexachlorocyclohexane (HCH), showing great application potential in the treatment of stubborn pollutants and suitable for environmental bioremediation and industrial catalysis.

The invention belongs to the technical field of enzyme engineering, and particularly relates to application of halogenated alkane dehalogenase in selective preparation of S-configuration aryl ethanol, the halogenated alkane dehalogenase is DbjA, and the amino acid sequence of the halogenated alkane dehalogenase is shown as SEQ ID NO.1. According to the invention, the halogenated alkane dehalogenase DbjA has stereoselectivity for the reaction of catalyzing phenyl chloride to prepare S-configuration aryl ethanol, the enantiomer selectivity for the catalytic reaction of different substrates can reach 500 or more, the ee value of the generated substrate can reach 99%, and meanwhile, the yield of 40% or more can be maintained; and a new solution is provided for the problems of complex operation and large workload in the preparation process of S-configuration aryl ethanol.

The invention discloses preparation and application of a protein composite imprinting material for targeted recognition of strigolactones. The method comprises the following steps: 1, fusing halogenated alkane dehalogenase into ShHTL7 for induced expression to obtain recombinant protein; 2, preparing nano magnetic ferroferric oxide hollow microspheres; 3, carrying out surface amination modification on the nano magnetic ferroferric oxide hollow microspheres; 4, functionalizing halogenated alkanoic acid on the surfaces of the nano magnetic ferroferric oxide hollow microspheres subjected to amination modification; and 5, preparing the strigolactones surface protein molecularly imprinted polymer. The protein composite imprinting material which is high in adsorption efficiency, high in adsorption selectivity and capable of being rapidly enriched is prepared, shows superparamagnetism, can be rapidly separated under the action of a magnet, shows ultrahigh targeted recognition capacity on strigolactones, can be used for targeted extraction of strigolactones in parasitic plant root exudates, and has a good application prospect. The materials are convenient and fast to recycle and can be repeatedly used.

The present application relates to the technical field of chemical genetics marking, in particular to a kind of probe for marking quantifying protein dynamics.The probe includes the compound of the following structural formula: the compound contains halogenated alkane chain, can be specifically covalently combined with halogenated alkane dehalogenase-HaloTag label protein.It is combined with the fusion protein containing HaloTag in cell with the purpose of protein, with high specificity and stability, low toxicity, with good biocompatibility and strong anti-photobleaching ability in live cell imaging.The compound has the nature of space limited emission effect, through HaloTag bridge, can sensitively mark and respond to the spatial dynamic change of any single protein molecule in cell physiological process, and then realize the quantification and visualization of protein dynamics in method, for recording physiological activity under different time scales.

The invention relates to the technical field of genetic engineering, in particular to an engineering bacterium capable of producing 2-halogenated acid dehalogenase at high yield as well as a construction method and application of the engineering bacterium. Trichoderma viride 2-halogenated acid dehalogenase gene TvHAD1 is over-expressed in escherichia coli or pichia pastoris, engineering bacteria of the 2-halogenated acid dehalogenase are constructed, and the engineering bacteria of the high-yield 2-halogenated acid dehalogenase are obtained through collaborative optimization of fermentation processes such as induction conditions and metal ions. And efficient heterologous expression and large-scale preparation of the enzyme are realized. Under the shake flask scale, the enzyme yield of the recombinant pichia pastoris engineering bacteria can reach 102.51 mg / L, the enzyme yield of the recombinant escherichia coli engineering bacteria is 21.07 mg / L, and high yield of the 2-halogenated acid dehalogenase is realized.

This invention relates to the fields of enzyme engineering and biocatalysis, disclosing a halohydrin dehalogenase mutant and its catalytic enantioselective dehalogenation hydroxylation reaction of a series of racemic ε-halohydrins, preparing corresponding chiral ε-diols and chiral ε-halohydrins. The invention utilizes a fungicide derived from *Agrobacterium radiodurans* (…). Agrobacterium tumefaciens The wild-type haloalcohol dehalogenase HheC (amino acid sequence SEQ ID NO:2) of AD1 was used as the parent enzyme, and was obtained by combining 2 to 6 amino acid site mutations at L142, W249, P84, T134, N176, and F186. The mutant can be used to catalyze the kinetic resolution and dehalogenation reactions of racemic ε-haloalcohols, synthesizing a series of high-optical-purity chiral ε-diols and chiral ε-haloalcohols. These compounds are important chiral building blocks for the preparation of various pharmaceutical chemicals and have broad industrial application value.

The application discloses an ancestral dehalogenase Ans171 and application thereof in degrading sulfur mustard halogenated pollutants. The amino acid sequence of the ancestral dehalogenase Ans171 is shown as SEQ ID NO. 2, and the nucleotide sequence of the coding gene is shown as SEQ ID NO. 3. The ancestral dehalogenase Ans171 of the application can efficiently degrade sulfur mustard simulant agent dichlorodiethyl sulfide. After 200 μL of crude enzyme solution of the ancestral dehalogenase Ans171 is added into 2 μL of dichlorodiethyl sulfide and reacted for 5 min, the degradation rate reaches 99.7%. The thermal stability and degradation efficiency of the ancestral dehalogenase Ans171 of the application are significantly higher than those of a modern enzyme, and the ancestral dehalogenase Ans171 has a good application prospect in bioremediation of sulfur mustard halogenated pollutants.

The invention relates to the technical field of enzyme engineering, namely biological catalysis, and discloses a halohydrin dehalogenase mutant and application of the halohydrin dehalogenase mutant to catalysis of enantioselective desymmetry of a series of 3-substituted oxetane and synthesis of corresponding chiral gamma-azido alcohol. Wild halohydrin dehalogenase (the amino acid sequence is SEQ ID NO: 2) derived from Ilumacter coccineus YM16-304 is used as a parent enzyme, and 2-5 amino acid site combination mutation is carried out on sites I104, Y18, T106, V105 and L103, so that the halohydrin dehalogenase is obtained. The chiral gamma-azido-alcohol is applied to catalysis of enantioselective azidation reaction of 3-substituted oxetane and sodium azide to synthesize a series of chiral gamma-azido-alcohol with high optical purity, and can be used as a key chiral intermediate to prepare chiral blocks such as triazole compounds, gamma-amino-alcohol and macrolactam. The method has application value in preparation of various bioactive drugs and fine chemicals.

The application relates to the technical field of inks, in particular to a high-flexibility acrylic epoxy resin for inks and a preparation method thereof. The preparation method of the high-flexibility acrylic epoxy resin for inks comprises the following steps: synthesizing an acrylic copolymer, carrying out a catalytic reaction of immobilized halogen alcohol dehalogenase, adding a flexible modifier, catalyzing by immobilized lipase and carrying out a grafting reaction of an epoxy resin. The application introduces long flexible side chains by enzyme-catalyzed open-loop grafting of flexible segments, improves the activity of molecular segments on the basis of retaining the original reaction activity and skeleton strength of the acrylic epoxy resin, improves the packing density of the long flexible side chains, improves the activity of molecular segments, and when the prepared acrylic epoxy resin is used in inks, the flexibility of ink films can be improved, the curing speed and surface hardness can be considered, meanwhile, the acrylic epoxy resin does not cause adverse effects on the transparency and storage stability of the ink films.