217 results about "Lyase" patented technology

The invention provides a kit for rapidly detecting MRSA and drug-resistant gene nucleic acid POCT of the MRSA based on a phage lyase mutant. The kit comprises nucleic acid extraction and purification and triple PCR rapid amplification detection reagents. The staphylococcus aureus bacteriophage lyase mutant nucleic acid release liquid containing the amino acid sequence as shown in SEQ ID NO: 1 is adopted, wall breaking can be rapidly conducted on staphylococcus aureus, and nucleic acid is released. The effect of the nucleic acid release liquid acting for 5 minutes is equivalent to or even better than that of the lysozyme acting for 60 minutes; according to the present invention, with the combination of the MRSA and drug-resistant gene triple PCR rapid amplification detection reagent, the total detection time is only 35 min, the time is saved by 55 min or more than 55 min compared with the lysozyme method, and the MRSA and drug-resistant gene nucleic acid POCT rapid detection can be achieved;

The invention belongs to the technical field of biology, and discloses a yarrowia lipolytica engineering strain for efficiently producing eicosapentaenoic acid based on citric acid metabolism regulation, a method and application of the yarrowia lipolytica engineering strain. Delta9 elongase, Delta8 desaturase, Delta5 desaturase and Delta17 desaturase genes in a fatty acid synthesis route, heterologous ATP-citrate lyase and a mitochondrial citric acid vector are overexpressed. According to the invention, ATP-citric acid lyase and a mitochondrial citric acid carrier gene which are respectively derived from Schizochylene sp. And Saccharomyces cerevisiae are jointly expressed in the yarrowia lipolytica engineering strain, so that the yields of EPA and grease in the yarrowia lipolytica engineering strain are remarkably improved, and compared with a control strain, the yields of the grease and the EPA are respectively improved by 243.35% and 161.85%.

The invention relates to the technical field of biological materials, in particular to a pH response collagen microneedle patch loaded with antibacterial nanoparticles as well as a preparation method and application of the pH response collagen microneedle patch loaded with the antibacterial nanoparticles, and the microneedle patch is prepared by loading the antibacterial nanoparticles, aldehyde modified F127 and recombinant three-type collagen COL3 on a polymer matrix. When the microneedle patch is applied to a chronic wound of a diabetic patient, the microneedle penetrates through the skin in a minimally invasive mode and directly reaches a wound focus area. Under the irradiation of 808nm near-infrared light, the mesoporous polydopamine absorbs light energy, so that the loaded lyase LysAB2 is quickly released, bacterial cell walls are specifically recognized and cracked, and bacteria are efficiently killed; meanwhile, the collagen hydrogel constructs an appropriate microenvironment at the wound, cell adhesion, proliferation and migration are promoted, and the wound healing process is accelerated. Compared with a traditional diabetes mellitus chronic bacterial infection wound treatment means, the microneedle patch provided by the invention provides an efficient and safe new strategy for treating the chronic bacterial infection wound of a diabetes mellitus patient.

The invention discloses a bacteriophage lyase and application thereof, and particularly relates to a bacteriophage lyase with an amino acid sequence as shown in SEQ INNO.2. The bacteriophage lyase can inhibit the growth of escherichia coli, staphylococcus and salmonella, can be used as an antibacterial substance in splitting gram-negative bacteria and can be used for preparing drugs for resisting gram-negative bacteria. The lyase can be used independently or compounded with other substances, the use concentration of antibiotics can be reduced by combining the lyase with the antibiotics, toxic and side effects (such as renal toxicity of polymyxin) of drugs are reduced, and meanwhile, generation of bacterial drug resistance is delayed. After the lyase and the chitosan are combined for use, remarkable antibacterial activity is generated, the antibacterial effect can be achieved without pre-treatment on bacteria, and the practicability is improved.

The invention provides low-temperature alginate lyase and application thereof, namely alginate lyase with low-temperature catalytic activity, which is separated from marine-derived cold monad bacteria, and the amino acid sequence of the alginate lyase is SEQ ID NO: 1; the amino acid sequence of the alginate lyase after the signal peptide is removed is SEQ ID NO: 3. The algin lyase Alg3889 provided by the invention has the optimal reaction temperature of about 10-16 DEG C, has good temperature stability, and still keeps more than 70% of enzyme activity after being incubated at 0-70 DEG C for 1 hour; the optimum pH value of the strain is 8.0, after the strain is incubated in a buffer system with the pH value of 3.0-10.6 for 12 h, about 90% of the highest enzyme activity can be kept, and the strain has high acid resistance and alkali resistance. The degradation product of the alginate oligosaccharide is alginate oligosaccharide with the polymerization degree of 2-6, and the alginate oligosaccharide has high application value.

The invention discloses a method for producing p-coumaric acid through double-enzyme cascade catalysis, which comprises the following steps: taking phenol, pyruvic acid and ammonium chloride as substrates, and carrying out cascade enzyme catalytic reaction on a tyrosine phenol lyase product and a tyrosine ammonia lyase product to synthesize p-coumaric acid. According to the method, phenol, pyruvic acid and ammonium chloride are used as substrates, a tyrosine phenol lyase product and a tyrosine ammonia lyase product are used as catalysts, the tyrosine phenol lyase product can convert the substrates into L-tyrosine, and the tyrosine ammonia lyase product can directly use the L-tyrosine as the substrate to convert the L-tyrosine into p-coumaric acid. Therefore, the tyrosine phenol lyase product and the tyrosine ammonia lyase product can realize cascade enzyme catalytic reaction in the same reactor, phenol with lower cost can be adopted as a conversion substrate, separation and purification of an intermediate product L-tyrosine are avoided, and the whole process flow is simple and efficient.

The invention relates to algin lyase FsAly7Z with high activity and high thermal stability, which is mainly used for producing brown algae trisaccharide, and an engineering bacterium and application of the algin lyase FsAly7Z. The invention provides an alginate lyase FsAly7Z derived from marine uncultured microorganisms and a preparation method of the alginate lyase FsAly7Z. The FsAly7Z has extremely high thermal stability, and compared with the enzyme activity loss of more than 50% after most enzymes are treated at 40 DEG C for 60 min, the FsAly7Z shows remarkably excellent thermal stability under the same condition, and the half-life period of the FsAly7Z exceeds 24 h; the FsAly7Z can be used for efficiently degrading sodium alginate and a polyG section which is difficult to degrade, and meanwhile, the FsAly7Z also shows the catalytic efficiency superior to that of most similar enzymes; and the content of brown algae trisaccharide in the enzymolysis product is high, so that the subsequent separation and purification process can be simplified, and the production cost can be reduced. In a word, the FsAly7Z is extremely high in thermal stability and excellent in catalytic efficiency, has both economical efficiency and conversion efficiency, can efficiently produce alginate oligosaccharide, is used for analyzing the structure and function association of alginate lyase, and has wide market prospects in the fields of medicine, food industry and the like.

The invention discloses a gelatin microcarrier lysate as well as a preparation method and application thereof. The gelatin microcarrier lysis buffer comprises lyase, a cell protective agent, a cell energy metabolism protective agent, an antioxidant, a buffer solution and a chelating agent. The gelatin microcarrier lysate has extremely strong protectiveness to cells, abandons damaging trypsin and EDTA, and innovatively adds a cell protection combination of a cell protective agent L-arginine, a cell energy metabolism protective agent sodium pyruvate and an antioxidant vitamin C, so that the cell survival rate (gt; 98%) and the retention rate (gt; the method is high in cell yield, good in safety and wide in range.

The invention provides an alginate lyase mutant Amut8C as well as a coding gene and application thereof, belongs to the technical field of gene engineering and enzyme engineering, and can solve the problem that existing alginate lyase cannot meet the requirements of severe reaction conditions such as high temperature and acidity in industrial application. The amino acid sequence of the alginate lyase mutant Amut8C disclosed by the invention is as shown in SEQ ID NO. 1. According to the space structure of the alginate lyase Algt, disulfide bonds in molecules are analyzed, the alginate lyase is mutated, and the obtained alginate lyase mutant Amut8C has heat resistance, acid and alkali resistance and digestive enzyme tolerance, and compared with the alginate lyase Algt, the alginate lyase mutant Amut8C has the advantages that the content of the alginate lyase mutant Amut8C is increased, and the content of the alginate lyase mutant Amut8C is increased. The alginate oligosaccharide with the polymerization degree of 1-4 prepared from the alginate lyase mutant Amut8C under the same enzymolysis condition is higher in yield. The method can be applied to the fields of feeds, foods, detergents or biofuels.

The invention relates to the isolation and characterisation of one or more aptamers against staphylococcal C-S lyase. The invention also provides aptamers, aptamer-conjugates, and minimal functional fragments thereof which may be used to inhibit the activity of said lyases and thereby inhibit Staphylococcus associated malodour.

The invention discloses an application of a tetrahydropyridopyrimidinone compound with a structural characteristic shown in a formula (I) or pharmaceutically acceptable salt, hydrate or crystal form thereof in treatment of human casein lyase (HsClpP) mediated diseases, and belongs to the field of chemical medicines. The technical problem to be solved by the invention is to provide the tetrahydropyridinopyrimidinone compound. The imidazolidine derivative is characterized by having a tetrahydropyridinopyrimidinone skeleton structure, and nitrogen atoms on an imidazolidine ring are connected with other substituent groups. The compounds have obvious activity of regulating and controlling HsClpP, and can be used for treating HsClpP-mediated related diseases.

The invention discloses recombinant saccharomyces cerevisiae for producing protopanoxadiol by metabolizing glycerol and a construction method thereof, and the construction method comprises the following steps: introducing a dammarendiol synthase DS gene expression cassette, a PPD synthase and reductase fusion protein PPDS-ATR1 gene expression cassette and a squalene oxidase ERG1 gene expression cassette into saccharomyces cerevisiae to obtain a recombinant bacterium 1; introducing a glycerol transport protein CjFPS1 gene expression cassette, a glycerol dehydrogenase OpGDH gene expression cassette, a dihydroxy acetone kinase DAK1 gene expression cassette and a 3-hydroxy-3-methylglutaryl coenzyme A reductase HMGR gene expression cassette into the recombinant bacterium 1 to obtain a recombinant bacterium 2; introducing citric acid lyase AnACLa and AnACLb gene expression cassettes and citric acid transporter protein YHM2 and CTP1 gene expression cassettes into the recombinant bacterium 2 to obtain a recombinant bacterium 3; experiments prove that the yield of the recombinant bacteria 2, 3PPD is 124 mg / L and 220 mg / L.

This invention provides the application of CTH inhibitors and / or xCT inhibitors in the preparation of drugs for inhibiting mammalian embryo implantation, relating to the field of pharmaceutical technology. This invention verifies the efficacy of cystathionine-γ-lyase (CTH) inhibitors and / or solute carrier family 7 member 11 (xCT) inhibitors in inhibiting mammalian embryo implantation. The results show that xCT inhibitors and / or CTH inhibitors can significantly inhibit the proliferation of luteinized granulosa cells and progesterone secretion in mouse models. Furthermore, in vivo injection of xCT inhibitors and / or CTH inhibitors into mouse models not only significantly inhibits ovulation and the maturation rate of released oocytes, but also significantly reduces the formation of functional corpus luteum and embryo implantation. Therefore, this application provides new ideas and methods for the development of contraceptive drugs.

The invention belongs to the technical field of enzyme gene engineering, and particularly relates to a methionine gamma-lyase mutant with improved enzyme activity and application thereof. The methionine gamma-lyase mutant CqMGLV160I / V265I with high activity is obtained by taking methionine gamma-lyase (CqMGL) which is derived from Calorama quimayensis and of which cysteine residues at 116 sites are replaced by histidine residues as a parent and carrying out site-specific mutagenesis on amino acid residues at key sites through transformation. The methionine gamma-lyase mutant CqMGLV160I / V265I with high activity can be obtained by using the methionine gamma-lyase mutant CqMGLV160I / V265I as the parent. By adopting the CqMGLV160I / V265I mutant disclosed by the invention, the conversion rate of enzyme catalysis can be improved, allicin is efficiently synthesized, and the mutant is successfully applied to the aspects of food, feed, chemical industry, medicine preparation and the like.

The invention discloses breeding of a pichia kluyverovii strain with high yield of beta-lyase and application of the pichia kluyverovii strain in beer fermentation, and belongs to the technical field of bioengineering. The pichia kluyveri B21 is preserved in the China General Microbiological Culture Collection Center, and the preservation number of the pichia kluyveri B21 is CGMCC (China General Microbiological Culture Collection Center) No.34028. The strain can produce high-yield beta-lyase, and the beta-lyase is catalyzed to produce volatile mercaptan and endows beer with fruity flavor. The contents of ethyl acetate, isoamyl acetate, isobutyl acetate, ethyl nonanoate, ethyl caprate, phenethyl acetate, linalool and nerol generated by the pichia kluyveri B21 in a hops simulated culture medium are relatively high, and the volatile substances can increase the fruit fragrance and flower fragrance of beer and enrich the flavor of the beer. The pichia kluyveri B21 is expected to be added into production of other food and beverages so as to enrich flavor substance components.

The invention relates to a lyase and a coding gene thereof, and application of the lyase in lysing bacteria. The bacteriophage capable of specifically cracking aeromonas hydrophila is obtained through separation and purification, lyase is cloned from a genome of the bacteriophage, and after prokaryotic expression and purification are carried out on the lyase, the obtained protein shows the characteristics of aeromonas hydrophila. The lyase disclosed by the invention has an obvious antibacterial effect on aeromonas hydrophila, so that the lyase has a potential application value in the aspect of controlling pollution of the aeromonas hydrophila.

This invention discloses methanogenic lyases with the same protein family annotation as PeiR lyases and their applications. These enzymes are obtained through homology screening based on the known characteristics of PeiR lyases targeting peptide bonds in the methanogenic cell wall, using the Pfam protein family database annotation. PeiR lyase proteins participate in the biological process of hydrolyzing archaea cell walls, effectively killing methanogens and reducing methane production. Proteins homologous to this protein have potential methane-reducing potential. This invention integrates rumen microbial metagenomic sequencing data, uses a series of bioinformatics software to screen methanogenic viral proteins, and combines Pfam functional domain annotation for homology analysis, ultimately identifying a series of lyases. These lyases were successfully expressed in a prokaryotic expression system, and in vitro gas production experiments also showed that the crude enzyme solution significantly reduced methane production.

The invention discloses a recombinant bacterium for producing L-lactic acid through co-utilization of glucose and xylose as well as a construction method and application of the recombinant bacterium. The invention relates to the technical field of biology, in particular to recombinant bacteria for producing L-lactic acid through co-utilization of glucose and xylose as well as a construction method and application. Methylglyoxal synthase, and / or pyruvate formate lyase, and / or fumarate reductase and / or L-lactic dehydrogenase in receptor Escherichia coli Rec1.0 are modified to obtain recombinant Escherichia coli, the recombinant Escherichia coli is fermented for 24 h, L-lactic acid is the only metabolite, and the yield reaches 2458 g / L; the consumption rate of xylose is 0.76 g / g DCW / h, the consumption rate of glucose is 0.69 g / g DCW / h, and the consumption rate of total sugar reaches 1.45 g / g DCW / h. The recombinant Escherichia coli constructed by the invention can realize efficient utilization of lignocellulose hydrolysate (containing glucose and xylose), and has a wide application prospect.

Methods and compositions relating to the engineering of modified proteins having homocysteine ​​activity are described. For example, a modified cystathionine-γ-lyase (CGL) comprising one or more amino acid substitutions and capable of degrading homocysteine ​​is disclosed. Furthermore, compositions and methods for treating homocysteine ​​depletion or hyperhomocysteinemia using the disclosed enzymes or nucleic acids are provided.

Aspects of the disclosure relate to aromatic amino acid ammonia lyases (ALs), phenylalanine ammonia lyases (PALs), and tyrosine ammonia lyase (TALs), including engineered enzymes, and their use in catalyzing chemical reactions.

The invention discloses a lactobacillus lactis for relieving constipation and a preparation method thereof, target strains are respectively separated from a fermented bean curd sample and screened by a pectinase screening agar culture medium, and are named as lactobacillus plantarum PD-14 and lactobacillus pentosus PD-16, the lactobacillus lactis has a gene for synthesizing pectate lyase, and the lactobacillus lactis has a gene for synthesizing pectate lyase. Relates to the microbial field. The invention provides a beneficial bacterium for relieving constipation and a preparation method thereof, wherein the beneficial bacterium is capable of efficiently relieving constipation, low in cost, stable and easy to industrialize.

This invention discloses a nicotine-resistant pectin lyase mutant and its application in tobacco leaf treatment. Using pectin lyase PL6 as the starting enzyme, this invention designs and optimizes mutation sites, designing single-site saturation mutations based on the two hydrogen bond formation sites between the enzyme and nicotine, Lys-107 and Asp-84, to construct mutants. The pectin lyase mutants D84A and K107R of this invention retain 83.63% and 81.30% of their enzyme activities at a 1% nicotine concentration, respectively. Spraying tobacco leaves with D84A and K107R enzyme solutions significantly improves tobacco leaf quality due to pectin degradation.

This invention discloses a methanogenic lyase homologous to PeiR lyase and its applications. The PeiR lyase protein, targeting the peptide bonds of the methanogenic cell wall, participates in the biological process of hydrolyzing the archaea cell wall, effectively killing methanogens and reducing methane production. Proteins homologous to this lyase possess potential methane-reducing potential. This invention utilizes a series of bioinformatics software to identify protein sequences encoded by methanogenic viruses from a large amount of rumen microbial virome sequencing data, and analyzes the structural homology between these proteins and PeiR. Ultimately, this invention successfully obtained a series of methanogenic lyases homologous to PeiR. To achieve the expression of these lyases, this invention designed multiple primers to synthesize the target sequences and successfully expressed them in a prokaryotic expression system. In vitro gas production experiments with the crude enzyme solution showed that these lyases are effective in reducing methane production.

The application discloses a kind of methyl aspartate lyase EcMAL mutant and its preparation method and application, belong to enzyme engineering technical field.The methyl aspartate lyase EcMAL mutant is any one of mutant EcMAL-M1, mutant EcMAL-M2, mutant EcMAL-M3 and mutant EcMAL-M4.The application is obtained methyl aspartate lyase EcMAL mutant by the mutation of the 329th, 331st, 361st and 365th amino acid sites of methyl aspartate lyase combined with high-throughput screening method.The mutant changes the previous recombinant enzyme only L-aspartic acid production status, so that it is possible to produce D-aspartic acid with methyl aspartate lyase as biological catalyst, and it is expected to be used in pharmaceutical industry.

To provide a modified tyrosine phenol lyase having improved activity for catalyzing a reaction (tyrosine cleavage reaction) for producing phenol from tyrosine.SOLUTION: Provided is a modified tyrosine phenol-lyase comprising an amino acid sequence in which one or more amino acid substitutions are introduced into an amino acid sequence of a wild-type tyrosine phenol-lyase, wherein an activity of catalyzing phenol production from tyrosine is improved compared to the wild-type tyrosine phenol-lyase, wherein the modified tyrosine phenol-lyase has amino acid substitutions at any one or more positions selected from positions corresponding to positions 25, 75, 144, 391, 419, and 456 of the amino acid sequence of SEQ ID NO: SELECTED DRAWING: Figure 1

This invention discloses a method for the efficient synthesis of gastrodin using recombinant Corynebacterium glutamicum, belonging to the fields of genetic engineering, fermentation engineering, and synthetic biology. This invention uses Corynebacterium glutamicum as the host bacterium and knocks out… pobA The gene was expressed, and the engineered strain WN01 / pB6 was constructed using the expression plasmid pEC-XK99E to express cladoid-pyruvate lyase, carboxylic acid reductase, and glycosyltransferase, achieving heterologous synthesis of gastrodin. Further overexpression was then performed. aroG Genes, overexpression of protein-coding genes that activate carboxylic acid reductase sfp The engineered strain WN05 / pB6 was able to synthesize 7.37 g / L of gastrodin at the shake flask level, and the gastrodin yield reached 17.77 g / L at 72 h of batch fermentation in a 5-L fermenter, with a yield of 0.247 g / L / h. This provides a brand-new biomanufacturing platform for the industrial production of gastrodin.

The application discloses a kind of high-yield O-succinyl-L-homoserine recombinant escherichia coli and its construction method and application, belong to genetic engineering and fermentation engineering technical field.The recombinant escherichia coli is expressed by expressing anti-feedback inhibition homoserine O-succinyltransferase metA fbr , aspartate ammonia-lyase aspA, aspartate transaminase aspC, 2 ketoglutarate decarboxylase sucA, dihydrothioctic acid succinyltransferase sucB and efflux protein yjeH, knock out metJ, improve intracellular NADPH availability, to enhance OSH synthesis pathway metabolic flow, precursor succinyl coenzyme A supply and product efflux.Using 5 L bioreactor for fed-batch fermentation, after fermentation 60 h, OSH production is as high as 137.4 g / L, and sugar acid conversion rate reaches 50.5%.The application provides engineering strain and technical realization path for the efficient fermentation of OSH.

This invention relates to recombinant lyases and variants thereof, which have improved stability or bactericidal activity relative to the recombinant lyases. This application also relates to pharmaceutical compositions comprising the recombinant lyases and variants thereof, and their use for the prevention and / or treatment of bacterial infections (e.g., bacterial vaginosis).

The present invention relates to a method for producing α-sialyl-β-D-galactoside saccharides, particularly α-sialyl-(2→3)-β-D-galactoside saccharides and α-sialyl-(2→6)-β-D-galactoside saccharides, from a β-D-galactoside saccharide, a sialic acid donor, and an enzyme with β-galactoside α-sialyltransferase activity. The enzymes with β-galactoside α-sialyltransferase activity used herein do not exhibit catalytic activity towards the hydrolysis of cytidine 5'-monophospho-N-acetyl-neuraminic acid to cytidine and N-acetyl-neuraminic acid. The method can be performed in vitro and in vivo using a genetically engineered cell comprising a nucleic acid encoding said enzyme. Further, said process may be adapted to produce the sialic acid donor CMP-Neu5Ac from low-cost substrates N-acetyl-D-glucosamine (GlcNAc), pyruvate, a cytidine phosphate (CMP, CDP or CTP) and polyphosphate in a single reaction mixture with a set of optionally immobilized or optionally co-immobilized enzymes comprising N-acylglucoamine 2-epimerase (AGE), an N-acetylneuraminate lyase (NAL), an N-acylneuraminate cytidylyltransferase (CSS), optional a uridine kinase (UDK), a uridine monophosphate kinase and a polyphosphate kinase 3 (PPK3).

This invention discloses an alkali-resistant xanthan gum side-chain lyase mutant and its preparation method, which is effective against alkali-resistant xanthan gum from Bacillus subtilis (Bacillus spp.). Paenibacillus nanensis Site-directed mutagenesis was performed on wild-type xanthan gum side-chain lyase to obtain alkali-resistant xanthan gum side-chain lyase mutants. The site-directed mutagenesis included at least one of Y260V, T387A, and I422S. The xanthan gum side-chain lyase mutants of this invention showed significantly increased enzyme activity compared to the original enzyme after treatment with an alkaline buffer system at 40°C, with the three mutants Y260V / T387A / I422S showing the greatest increase. These mutants are suitable for large-scale degradation of xanthan gum side chains under alkaline conditions, possessing significant social and economic benefits and broad application prospects.