54 results about "Cutinase" patented technology

The invention discloses a composite microbial enzyme preparation and a preparation method thereof, and belongs to the technical field of bioplastic degradation. The enzyme preparation is synergistically constructed through a temperature-sensitive MOFs / Schiff base network composite carrier and black phosphorus quantum dots, FeO (at) ZIF-8 is taken as a core, and PNIPAM is grafted on the surface to form temperature-responsive MOFs; constructing a three-dimensional network carrier through Schiff base crosslinking of the CMC and the CNF; the complex enzyme is loaded on MOFs pore channels and then is embedded in a carrier network together with the black phosphorus quantum dots. The temperature-sensitive MOFs open pore channels to release enzyme when the compost is heated, and the Schiff base network dissociates secondary released enzyme under an acidic condition; the black phosphorus quantum dots promote enzyme activity and strengthen carrier response through a photothermal effect; the compound enzyme contains cutinase, alkaline lipase, neutral lipase, alpha-amylase, saccharifying enzyme, cellulase, laccase and protease, and can synergistically degrade PLA, PBAT and starch-based plastic. According to the invention, dual environment response controlled release, photo-thermal synergistic catalysis and synergistic degradation of the enzyme preparation are realized, and the plastic degradation period is shortened under the composting condition.

The invention discloses keratinase Phyto-Keratinase and application of the keratinase Phyto-Keratinase. The keratinase Phyto-Keratinase comprises the following components: (1) protease which is coded by a nucleotide sequence as shown in SEQ ID NO.1; and / or (2) protease which is derived from bacillus subtilis, is coded by a nucleic acid sequence with more than 98% of identity with the SEQ ID NO.1 nucleotide sequence and has the characteristic of hydrolyzing keratin. According to the plant keratinase Phyto-Keratinase disclosed by the invention, the Phyto-Keratinase (plant keratinase) produced by utilizing microbial fermentation is a serine proteolytic enzyme, is also a biocompatible cutinase, and is obtained by fermenting saccharomyces cerevisiae. Phyto-Keratinase provides a nitrogen source for the growth of plants (beans), and contains various amino acids (functional peptides) and special proteolytic enzymes derived from the plants at the same time. The Phyto-keratinase can provide the effects of resisting inflammation, removing cutin and providing nutrition for skin regeneration at the same time, can relieve various inflammations by inhibiting PAR-2 receptors, and has potential value in the fields of cosmetics, beauty medicine and environment.

Leaf-branch compost cutinase (LCC) and other cutinase mutants with activity over ranges of temperature and pH are provided.

The invention discloses a fungal cutinase CpL-5Ct1 and an application of the fungal cutinase CpL-5Ct1 in degradation of polyurethane plastics. When the cutinase CpL-5Ct1 is used for degrading the PU foam, the shape of the PU foam can be obviously changed within 3 days at the temperature of 55 DEG C, and the PU foam is in a broken powder shape; the plastic biological enzyme method depolymerization technology is mild in condition, efficient in process, few in by-product and environmentally friendly, is an ideal means for waste plastic treatment, and has important application value in the aspects of waste PU plastic treatment and resource utilization.

The invention belongs to the technical field of enzyme engineering, and discloses a high-activity and high-stability PET hydrolase mutant and application thereof. The high-activity and high-stability PET hydrolase mutant is a protein A, a protein B or a protein C, wherein the protein A is obtained by mutating the 132 amino acid of wild cutinase TfCut from glutamine Q to lysine K; the protein B is obtained by mutating the 244th and 293rd amino acids into cysteine C on the basis of the protein A; protein C is a fusion protein containing protein A or protein B. The modified enzyme breaks through a trade-off mechanism of enzyme catalytic activity and stability, and the activity and thermal stability of the modified enzyme are greatly improved, so that the industrial application value of the PET hydrolase in the aspect of PET degradation is improved.

The invention discloses a method for improving cutinase catalytic degradation of PET (Polyethylene Terephthalate) plastics, which comprises the following steps: preparing a macromolecular condensed phase, and collecting a concentrated phase of the condensed phase in a centrifugal manner; adding an LCC-ICCG-218Y enzyme solution and a PET solid into the concentrated phase, and oscillating to enable the LCC-ICCG-218Y enzyme solution and the PET solid to be uniformly distributed in a reaction system; reacting in an oil bath at 40-72 DEG C, collecting a reaction solution, and characterizing a degradation product through ultraviolet rays. Compared with a phosphate buffer solution, the method has the advantage that the degradation rate of the PET plastic at 40 DEG C and 50 DEG C is finally doubled compared with that of a buffer solution system. According to the method, a simple method for improving enzymolysis of the PET plastic is established, only the reaction microenvironment of the enzyme is regulated and controlled, complex protein modification and enzyme immobilization means are not needed, and the method has certain application value.

The invention discloses low-temperature cutinase CtCut derived from marine fungi and capable of efficiently degrading PU and PET plastics and application of the low-temperature cutinase CtCut. The amino acid sequence of the low-temperature cutinase CtCut is shown as SEQ ID NO. 2, and the nucleotide sequence of the low-temperature cutinase CtCut is shown as SEQ ID NO. 1. The CtCut can degrade various polyurethanes (PU) such as polyester type PU ImpranilTMDLN, polyether type PU foam and PBA-PU at the temperature of 35 DEG C, and also has a degradation effect on polyethylene glycol terephthalate (PET). The CtCut can efficiently break ester bonds in PET, ester bonds in a PU soft segment and carbamate bonds in a PU hard segment. The cutinase CtCut has substrate heterogeneity, can efficiently degrade PET (Polyethylene Terephthalate) and PU (Polyurethane) plastics, and can provide theoretical guidance and technical support for recycling various types of PU and mixed plastics in the future.

The invention belongs to the technical field of polyester surface modification, and relates to a method for promoting cutinase to hydrolyze polyester by using triton. The method comprises the following steps: adding triton into a Tris-HCl buffer solution, uniformly mixing, and adjusting the pH value with hydrochloric acid to obtain a triton buffer solution; and immersing the refined terylene into a triton buffer solution, adding a cutinase preparation for hydrolysis reaction, inactivating cutinase, taking out the terylene, washing and drying. Triton is used as an additive to improve the activity and stability of cutinase, promote the hydrolysis effect of cutinase on polyester, and increase the hydrolysis release product by 229.8%, so that cutinase can hydrolyze polyester with higher crystallinity.

The invention relates to cutinase as well as a coding gene and application thereof. The amino acid sequence of the cutinase SCC is as shown in SEQ ID NO. 1, and the nucleotide sequence of the cutinase SCC is as shown in SEQ ID NO. 2. The inventor of the application finds out a novel cutinase SCC of a serine hydrolase family from Saccharothrix carnea, and the cutinase SCC is easy for heterologous expression and purification, and can efficiently and specifically catalyze and degrade mono (2-hydroxyethyl) terephthalate (MHET) and polyethylene glycol terephthalate (PET). And compared with the existing MHET degrading enzyme, the MHET degrading enzyme has the advantages that the catalytic efficiency is obviously improved, the use amount of the enzyme can be effectively reduced in the process of catalytically degrading MHET and PET, and the use cost of the enzyme is reduced.

The application discloses a controllable preparation method of high-activity earthworm protein peptide and belongs to the technical field of earthworm processing. The preparation method comprises the following steps: (1) adding amylase and cutinase to earthworm paste for hydrolysis; (2) filtering by using a filter; (3) hydrolyzing the filtrate by using a composite enzyme; (4) adding alkaline protease to the filter residue for hydrolysis, and then adding high-temperature protease for hydrolysis; (5) filtering the hydrolyzed paste, and then filtering and treating the paste by using a ceramic microfilter to obtain low-molecular-weight earthworm protein peptide; and (6) mixing the obtained earthworm water-soluble protein hydrolysate and the obtained earthworm insoluble protein polypeptide, and then concentrating and drying to obtain high-activity earthworm protein peptide. According to the application, the water-soluble protein and the water-insoluble protein in the earthworm are step-by-step differentially hydrolyzed, the protein resources in the earthworm are maximally utilized, the polypeptide / protein yield can be more than 85%, waste is avoided, and the activity of the obtained protein peptide is ensured.

The invention discloses a leaf and branch compost cutinase link and application thereof, and belongs to the technical field of enzyme molecule construction. The leaf-branch compost cutinase kink is obtained by adjusting the connection sequence of fragments in leaf-branch compost cutinase, and comprises a fragment II, a connecting peptide fragment I, a fragment I, a connecting peptide fragment II and a fragment III which are connected in sequence from the N end to the C end; wherein the fragment I, the fragment II and the fragment III respectively correspond to 37th to 79th amino acids, 81st to 143rd amino acids and 150th to 293rd amino acids or homologous sequences thereof of the leaf branch compost cutinase. According to the method, protein topology engineering and an artificial intelligence assisted protein design technology are organically combined, knot topology transformation is performed on the leaf branch compost cutinase, sequence optimization is performed on a connecting peptide fragment of the knot, and the leaf branch compost cutinase knot with good biological activity is obtained; the expression quantity and the stability of the leaf and branch compost cutinase kink are improved, the problem of kinetic barriers existing in protein kink combination is solved, and the method has important application value.

The invention belongs to the technical field of degradation of waste textiles, and relates to a method for improving enzymolysis efficiency of polyester through pretreatment of an alkaline eutectic solvent, which comprises the following steps: mixing a hydrogen bond donor and a hydrogen bond acceptor, heating and stirring until the mixture is clear and transparent to obtain the eutectic solvent; the preparation method comprises the following steps: soaking terylene in a deep eutectic solvent, separating out terylene, and washing with ethanol and deionized water until the pH is neutral, so as to obtain pretreated terylene; cutinase is dissolved in a Tris-HCl buffer solution to obtain an enzymolysis solution, pretreated polyester is immersed for a reaction, and then inactivation is performed. The deep-eutectic solvent is adopted for promoting cutinase to conduct enzymolysis on the polyester, the deep-eutectic solvent has the advantages of being simple in preparation method, low in cost, biodegradable, recyclable and non-toxic, the crystallization degree of the polyester is reduced through the CHCl / EA deep-eutectic solvent, then enzymolysis is conducted, and the total product release amount of enzyme hydrolysis is increased by 5 times.

The present invention discloses a Thermobifida fusca cutinase mutant and a method for its soluble expression, belonging to the technical field of enzyme engineering. In the present invention, the mutant D204C / E253C and the periplasmic protein disulfide isomerase DsbC are co-expressed in the Escherichia coli mutant strain E. coli Origami B(DE3). However, during the expression process of the obtained recombinant strain E. coli Origami B(DE3) / pSCDsbC-D204C / E253C, it is prone to misfolding to form a large number of inclusion bodies, and the soluble expression ratio is extremely low. The present invention further realizes the efficient soluble expression of the cutinase mutant D204C / E253C through co-expression with the molecular chaperone protein DsbC in the Escherichia coli mutant strain E. coli Origami B(DE3), and has certain industrial application prospects.

The invention relates to the technical field of enzyme engineering and recombinant protein expression, and discloses a mutant for improving cutinase yield and a preparation method thereof. The mutant protein is based on an amino acid sequence as shown in SEQ ID NO.1 and has G93Q or H209F single-point substitution, and the site number takes first methionine in the SEQ ID NO.1 as the first site. The invention also provides a nucleic acid molecule for coding the mutant, a recombinant expression vector and a recombinant host cell, and a method for adding an inducer to induce expression when OD600 is 0.6-1.2 in a culture process so as to obtain a culture containing the mutant. Embodiments show that the enzyme activity per unit volume of the mutant culture is higher than that of a control group.

The invention discloses a novel straw cutin removal method, and belongs to the technical field of agricultural waste treatment. According to the method, the hydrolytic acidification is matched with the biological enzyme deconstruction technology, especially the coexpression transporter system cutinase, so that the cutin component in the straw is effectively removed, and the biodegradability and the subsequent utilization value of the straw are improved.

The present invention relates to a method for treating a textile comprising a polyester with low crystallinity by contacting the textile with a cutinase, as well as a textile produced by such method.

The application discloses a cutinase mutant and application thereof to efficient degradation of PET. The amino acid sequence of the cutinase mutant is shown in SEQ ID NO. 2, and is obtained by replacing phenylalanine at the 239th amino acid in the amino acid sequence of a wild-type cutinase shown in SEQ ID NO. 1 with alanine. The cutinase mutant is expressed in E. coli, and compared with the wild-type enzyme, the activity of the mutant enzyme is increased in stability, the catalytic efficiency is improved, and the yield of a PET plastic (polyethylene terephthalate, PET) degradation product is increased by 42.6 times. The improved cutinase mutant improves the PET degradation efficiency, reduces the production cost, and is more suitable for industrial application.

The invention belongs to the technical field of papermaking, and discloses a method for preparing a hybrid wood chemical mechanical pulp assisted by a deep-sea composite enzyme. Method: 1) soaking the hybrid wood chips in water, steaming, adding alkaline hot water, adjusting the pH to 8-9, adding a deep-sea composite enzyme preparation for treatment; the deep-sea composite enzyme preparation is a mixed enzyme of cellulase, xylanase and cutinase; 2) adding an alkali solution with a mass concentration of 4-10% to the above-mentioned slurry for treatment, refining, hot water dilution and adjusting the pH to 7-9, adding a deep-sea composite enzyme preparation, treating the slurry at 70-80°C for 30-90min, screening, beating, and obtaining a hybrid wood chemical mechanical pulp; the composite enzyme preparation is a mixed enzyme of three enzymes of cellulase, xylanase and lignin oxidase. The method of the present invention is simple, reduces pulping energy consumption, and improves pulping performance, and has good practical application value and industrial application prospects.

The present invention discloses a fungal cutinase CpLCut1 capable of degrading polyester plastics and its application. The present invention provides a cutinase encoding gene capable of degrading polyurethane plastics, the nucleotide sequence of which is SEQ ID NO.1, and the amino acid sequence of the encoded cutinase protein is SEQ ID NO.2. The recombinant cutinase obtained by the engineered strain constructed using the gene uses 4-nitrophenylbutyrate as a substrate, and the specific enzyme activity can reach 52.94 U / mg. The cutinase can destroy the structure of polyurethane plastics and produce a clear transparent hydrolysis zone on a flat plate. The cutinase can degrade PUR foam, and a significant change in the morphology of the PU foam can be clearly observed within 3 hours at 55°C, presenting a broken powder. Therefore, the use of the cutinase encoding gene as an element can be widely applied to the degradation and resource utilization of waste polyester plastics.

This invention discloses a BhrPETase keratinase mutant and its application in the degradation of polyethylene terephthalate (PET), belonging to the fields of environmental science and enzyme engineering. The site numbers are based on the amino acid sequence shown in SEQ ID NO:1. The mutant contains substitutions of S14G, H78Y, and S184G relative to SEQ ID NO:1, with the preferred amino acid sequence shown in SEQ ID NO:9. Under conditions of 1 mg enzyme protein / g substrate, 65°C, and 96 h, the preferred mutant exhibits a degradation rate of 90.4% for PET membranes and a half-life of 150 h at 60°C, making it suitable for enzymatic degradation and depolymerization of PET.

The invention discloses a cutinase mutant and application thereof in degradation of polyethylene glycol terephthalate, and belongs to the field of environmental science. According to the present invention, BhrPETase derived from bacteria HR29 and LC cutinase of a metagenome in plant compost are modified to obtain mutants, the mutants are subjected to single-point or multi-point mutation near the substrate binding site of the BhrPETase or the LC cutinase, and the 184th site and the 156th site are subjected to single-point mutation or combined mutation to construct six mutants; compared with a wild type enzyme, the six mutants (M1-M6) have the advantages that the enzyme activity and the PET degradation efficiency are obviously improved, and the industrial prospect is good.

A cutinase is provided in the present invention. In addition, compositions comprising cutinase and methods of making and using cutinase are provided.

The present disclosure provides a method for rapid and harmless treatment of a livestock and poultry carcass / residue by a bacterium-enzyme complex and relates to the technical field of dead livestock and poultry treatment. In the present disclosure, a composite microbial inoculant is compounded from protease-, lipase-, and keratinase-producing strains, and can be applied to the degradation of fats and proteins as well as deodorization. The composite microbial inoculant is further combined with a protease preparation and a lipase preparation to prepare the bacterium-enzyme complex. The bacterium-enzyme complex can quickly and harmlessly treat the carcasses / residues of pig, cattle, sheep, and chicken, and has a short treatment period, mild and controllable reaction conditions, and safe and non-toxic reaction process and reaction products. During the reaction, harmful gases such as CH4 and NH3 and greenhouse gases such as CO2 released by the method are less than those by other methods.

The application discloses a wool tricarboxylic acid blended fabric and a preparation method thereof, and belongs to the technical field of textile printing and dyeing. The method comprises the following steps: respectively performing normal-pressure low-temperature plasma pretreatment and specific enzyme synergistic modification on wool sliver and tricarboxylic acid short fiber sliver, moderately passivating wool scales by using protease, and promoting surface micro-deacetylation of tricarboxylic acid by using esterase or cutinase; mixing the slivers, spinning and weaving into a fabric; performing one-bath same-color dyeing by using disperse / acid dyes at 100 DEG C under normal pressure; and performing medium-low temperature heat setting. Through synergistic effect of plasma physical etching and biological enzyme chemical degradation, the technical prejudice that tricarboxylic acid needs high-temperature dyeing at 130 DEG C is broken. The method can obtain excellent same color under normal pressure, significantly improves wool strength retention rate, avoids wool heat damage and yellowing, the finished product has low shrinkage rate after machine washing, is green and environment-friendly, and is suitable for industrial popularization.

The present invention relates to a method for treating a textile comprising a polyester with low crystallinity by contacting the textile with a cutinase, as well as a textile produced by such method.

The invention discloses a coding gene of cutinase CutBJB6 for synthesizing a plurality of fatty acid ethyl esters from a Lacnella arida source and an application of the coding gene of cutinase CutBJB6. The amino acid sequence of the cutinase CutBJB6 is as shown in SEQ ID NO. 1, and the nucleotide sequence of the coding gene of the cutinase CutBJB6 is as shown in SEQ ID NO. 2. The invention belongs to the technical field of biosynthesis, and particularly discloses a coding gene of the Lacnellada-derived cutinase CutBJB6 and construction of an escherichia coli expression vector carrying the gene, and also discloses application of the Lacnellada-derived cutinase CutBJB6 in catalytic synthesis of ethyl butyrate, ethyl valerate, ethyl hexanoate, ethyl caprylate and ethyl caprate in a water-phase system. The invention further discloses a preparation method of the Lacnellada-derived cutinase CutBJB6 and application of the Lacnellada-derived cutinase CutBJB6 in the field of biosynthesis of the Lacnellada-derived cutinase CutBJB6. The invention firstly provides the cutinase CutBJB6 which has the capability of synthesizing fatty acid ethyl ester in a water phase system and is derived from the Lacnella arida and the coding gene of the cutinase CutBJB6.

The application discloses a cutinase capable of degrading polyester plastic and application thereof. The application provides a cutinase gene capable of degrading polyester plastic, wherein the nucleotide sequence of the cutinase gene is SEQ ID NO. 1, and the amino acid sequence of the coded cutinase protein is SEQ ID NO. 2. The cutinase can destroy the complete structure of plastic, and the surface of the plastic appears holes, cracks and other erosion. In addition, the cutinase can degrade different types of polyester plastic, wherein the weight loss rates of PUR foam and polyester plastic PCL reach 33.88% and 83.44% respectively within 2 days, and the degradation rates of agricultural degradable mulch PBAT and PU plastic synthetic oligomer PBA-PU reach 66.70% and 64.41% respectively within 4 days. The cutinase gene can be widely applied to degradation and resource utilization of waste polyester plastic as an element.

The present invention belongs to the fields of enzyme engineering and plastic degradation technology, and specifically relates to a cutinase variant, a preparation method thereof, and its application in plastic degradation. The present invention uses cutinase ICCG as a starting enzyme and utilizes bioinformatics and protein engineering techniques to engineer a series of mutants with enhanced affinity for the substrate PET. Experimental verification shows that the obtained cutinase variants have a higher affinity for PET and higher hydrolytic activity for PET, hydrolyzing 90% of the PET substrate in just 3.7 hours or less. Furthermore, over 99% of the hydrolysis products are terephthalic acid and ethylene glycol monomers, with virtually no incompletely degraded BHET and MHET. This more thorough hydrolysis process is therefore more conducive to meeting the practical application needs of depolymerization and reuse of PET waste, and therefore has excellent practical application value.

Presented herein is an artificial cutinase polypeptide in which the first C31-C109 disulfide bridge is deleted, resulting in an increase in enzymatic activity. In certain embodiments C31 is mutated to an Alanine or a Serine. Further presented are means of creating and using said cutinase for the purposes of degrading PET.

The invention discloses cutinase ChCut1 with a function of degrading polyurethane plastics. The amino acid sequence of the cutinase is shown as SEQ ID NO. 1. The invention also provides an extracellular expression method and application of the cutinase. The cutinase can be massively expressed in the saccharomyces cerevisiae, so that the efficient, stable and low-cost cutinase is produced. The cutinase can effectively degrade Impranil PU (polyurethane) under the condition of 28 DEG C, so that a plastic polymer structure is destroyed, and biodegradation under a mild condition is realized.