124 results about "Lyase" patented technology

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.

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 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.

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.

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.

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.

This invention relates to the fields of synthetic biology and biochemical engineering, and provides an engineered *Yersinia lipolytica* strain that enhances coumaric acid accumulation, its construction method, and its applications. The engineered *Yersinia lipolytica* strain of this invention uses *Yersinia lipolytica* as the substrate strain, expresses the tyrosine ammonia-lyase encoding gene RgTAL derived from *Rhodotorula glutinis*, and completely knocks out the aminotransferase encoding gene containing the nucleotide sequence shown in SEQ ID NO 1 (database number YALI0A08734g). RgTAL converts endogenous L-tyrosine to coumaric acid. This invention reduces dependence on complex metabolic regulation and increases the coumaric acid accumulation level in *Yersinia lipolytica*.

The invention discloses application of bacteriophage lyase LysFEC14 in preparation of a composition for resisting gram-negative bacteria. The amino acid sequence of the bacteriophage lyase LysFEC14 is as shown in SEQ ID NO.01. The invention further discloses a preparation method of the bacteriophage lyase LysFEC14. The antibacterial composition disclosed by the invention shows high-efficiency cracking activity on various escherichia coli standard strains, still has a remarkable antibacterial effect at low concentration, and provides a green and efficient novel antibacterial composition for preventing and treating gram-negative bacterium infection, particularly drug-resistant escherichia coli infection.

The application 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, wherein a chassis strain of the engineering strain is Yarrowia lipolytica Po1f, and genes of Δ9 elongase, Δ8 desaturase, Δ5 desaturase and Δ17 desaturase in a fatty acid synthesis pathway, a heterologous ATP-citrate lyase and a mitochondrial citrate carrier are overexpressed. ATP-citrate lyases and mitochondrial citrate carrier genes respectively derived from Schizochytrium sp. and Saccharomyces cerevisiae are combined to be expressed in the Yarrowia lipolytica engineering strain, so that the production of EPA and oil in the Yarrowia lipolytica engineering strain is significantly improved, and compared with a control strain, the production of oil and EPA is respectively increased by 243.35% and 161.85%.

The invention discloses a method for optimizing fermentation of Thermus thermophilus to produce photolyase at high yield. The method comprises the following steps: S1, tank preparation; s2, preparing a culture medium; s3, inoculation: inoculating the culture medium with the Thermus thermophilus seed solution; s4, in 0-6 h, air is introduced at the temperature of 50-60 DEG C under the condition that the dissolved oxygen is 75-85%; in 6-6.5 h, the temperature is 70-80 DEG C, and the dissolved oxygen is 25-35%; in 6.5-12h, the temperature is 70-80 DEG C, the dissolved oxygen is 25-35%, air and pure oxygen are introduced, and 405nm blue light irradiation is carried out; 12-12.5 h: cooling to 25-35 DEG C, and stopping introducing gas to obtain final fermentation liquor; and S5, centrifuging the final fermentation liquor, collecting thalli, resuspending with PBS, and then sequentially performing heat sink, ice bath, secondary centrifugation and filtration to obtain clarified enzyme liquor. The method provided by the invention adopts a low-temperature high-oxygen-step heating-light induction fermentation mode to ferment the thermophilic thermophilic bacteria, and has the characteristics of improving the fermentation level, increasing the biological quality and increasing the amount of photolyase.

The application discloses an ammonia lyase mutant, a recombinant bacterium and application of the ammonia lyase mutant in preparation of aromatic L-amino acid, and relates to the technical field of bioengineering and enzyme catalysis. Compared with a wild-type ammonia lyase, the ammonia lyase mutant comprises any one of the following mutations: M152A and / or L149G, and the amino acid sequence of the wild-type ammonia lyase is shown as SEQ ID NO. 1. The application provides the ammonia lyase mutant with higher enzyme activity and catalytic efficiency than the wild type, which can catalyze preparation of aromatic L-amino acid with high selectivity, high stability and high conversion rate. The ammonia lyase mutant has high stereoselectivity, avoids tedious chiral separation, and can directly obtain a medical-grade high-purity product. The method for preparing aromatic L-amino acid provided by the application can reduce the cost of materials, does not need a cosolvent and other reagents, simplifies the preparation process of aromatic L-amino acid, and has good industrial application potential.

An immobilized enzyme fiber contactor includes a plurality of fibers disposed within a hollow conduit. The fibers have attached thereto an enzyme selected from the group consisting of an oxidoreductase, a transferase, a hydrolase, a lyase, an isomerase or a ligase. The enzyme may be attached to the fiber via an anchor group and optionally a bifunctional crosslinking agent. The enzyme may be applied, stripped, and re-applied without detaching the reactor or discarding the fibers. The immobilized enzyme fiber contactor can be used for carrying out a two-phase or single-phase enzymatic reaction.

The application discloses a high-yield beta-lyase Pichia kluyveri strain breeding and application in beer fermentation, belonging to the field of bioengineering technology. Pichia kluyveri The Pichia kluyveri (Pichia kluyveri) B21 is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 34028. The strain can produce high-yield beta-lyase, the enzyme catalyzes the production of volatile mercaptans, and can impart beer fruity aroma. The Pichia kluyveri B21 produces high content of ethyl acetate, isoamyl acetate, isobutyl acetate, ethyl nonanoate, ethyl decanoate, phenethyl acetate, linalool and nerol in hop simulation culture medium, and these volatile substances can increase beer fruity aroma, floral aroma and rich beer flavor. The Pichia kluyveri B21 is expected to be added to the production of other food and beverage to enrich flavor substance components.

The invention relates to a composition. The present invention relates to a solid carrier, an engineered phenylalanine ammonia lyase or a fragment thereof immobilized on the surface of the solid carrier, and a protective layer for protecting the engineered phenylalanine ammonia lyase or the fragment thereof by embedding the engineered phenylalanine ammonia lyase or the fragment thereof, and a functional component fixed on the surface of the protective layer, in which the functional component fixed on the surface of the protective layer is a polymer comprising repeating units, in which each repeating unit comprises at least one amino group and / or at least one mercapto group. The invention also relates to a method for producing said composition and to the use thereof.

The invention belongs to the field of biological enzyme engineering, and particularly relates to a high-stability alginate lyase mutant and a preparation method thereof. The invention relates to algin lyase mutants AncAlyA1 and AncAlyA2 with high stability. The algin lyase mutants AncAlyA1 and AncAlyA2 are characterized in that coded protein sequences of the algin lyase mutants AncAlyA1 and AncAlyA2 are shown as SEQ ID NO.2 and SEQ ID NO.3. The algin lyase is optimized through an ancestor sequence reconstruction technology, the activity of the optimized enzyme is remarkably improved, and algin degradation can be more efficiently catalyzed. And the heat stability and pH adaptability are enhanced, so that the enzyme keeps activity under wider temperature and pH conditions, the method adapts to complex environmental conditions in industrial production, and the performance reduction caused by environmental fluctuation is reduced. In addition, by optimizing the fermentation process, the enzyme yield is greatly increased, the production cost is reduced, and the economic benefit is improved. Due to the improvements, the technology is superior to the prior art in the aspects of enzyme catalysis efficiency, stability, adaptability and yield, and a better solution is provided for efficient utilization and industrial production of algin.

The invention provides a carbon chain elongase as well as a coding gene and application thereof, the amino acid sequence of the carbon chain elongase is as shown in a sequence table SEQ ID NO.1, the nucleotide sequence of the gene for coding the carbon chain elongase is as shown in a sequence table SEQ ID NO.2, and the carbon chain elongase can be used as a formaldehyde lyase and has a function of efficiently catalyzing formaldehyde to synthesize 1, 3-dihydroxyacetone.

The invention discloses an alkali-resistant xanthan gum side chain lyase mutant and a preparation method thereof, the alkali-resistant xanthan gum side chain lyase mutant is obtained by carrying out site-specific mutagenesis on wild type xanthan gum side chain lyase from paenibacillus nanensis, and the mutation site of the site-specific mutagenesis comprises at least one of Y260V, T387A and I422S. According to the xanthan gum side chain lyase mutant disclosed by the invention, the enzyme activity of the mutant treated by using an alkaline buffer system is obviously improved compared with that of an original enzyme at 40 DEG C, and the enzyme activity is improved to the greatest extent by using the three mutants Y260V / T387A / I422S, so that the xanthan gum side chain lyase mutant is suitable for large-scale degradation of a xanthan gum side chain under an alkaline condition, has important social and economic benefits and is wide in application prospect.

The application discloses a kind of alginate lyase mutants, coding gene, recombination expression vector and genetically engineered bacteria, belong to genetic engineering technical field.The application uses alginate lyase PpAly7A as starting template, and obtains the mutant with relatively specific activity and thermal stability by site-directed mutagenesis and combination mutation.The relative specific activity of mutant C169A, C66A / C169A, C66A / C169A / C214A, E87Q is increased to 131.42%, 162.89%, 160.97%, 133.82% respectively compared with wild enzyme PpAly7A.The destruction of Glu180-Arg178 salt bond (E180Q) improves thermal stability, and the half-life of the mutant at 50 DEG C is increased to 7.246d compared with wild enzyme PpAly7A, and the site can be used as a new target for subsequent rational design.

The invention belongs to the technical field of gene engineering and enzyme engineering, and aims to provide a tyrosine phenol lyase mutant which is higher in catalytic efficiency and more suitable for industrial production of levodopa and is derived from citrobacter freundii, and compared with a wild tyrosine phenol lyase type, the tyrosine phenol lyase mutant has T49R or / and M379R mutation; the amino acid sequence of the tyrosine phenol lyase wild type is shown as SEQ ID NO.2 in a sequence table. The mutant and the engineering strain provided by the invention have the huge advantages of high catalytic efficiency and high product yield.

The application discloses a methyl bacillus subtilis sulfoxide synthase MpEgtB mutant and application thereof, and belongs to the technical field of bioengineering. The application provides a methyl bacillus subutilis-derived sulfoxide synthase MpEgtB mutant capable of efficiently preparing ergothioneine and a modification method of the mutant, and the mutant protein is used to catalyze histidine trimethyl inner salt, L-Cys is used as a sulfur donor to directly convert the histidine trimethyl inner salt into histidine trimethyl inner salt-based cysteine sulfoxide, and the obtained mutant is coupled with Mycobacterium smegmatis-derived MsEgtD and MsEgtE to prepare L-EGT by using L-His and L-Met as substrates. The optimal mutant strain is used as a biological catalyst to perform whole-cell conversion, a continuous feeding strategy is adopted in a 5L fermenter, and the conversion is performed for 48h, so that the yield of L-EGT reaches 8.32g / L. The production process is simple, has few impurities, and has an important industrial application prospect.

The application belongs to the technical field of biochemical synthesis, and particularly relates to a high stereoselectivity amino-cleaving enzyme and a synthesis method of (S)-3-aminobutyronitrile and a salt thereof. The high stereoselectivity amino-cleaving enzyme is selected from the amino acid sequences shown in SEQ ID NO. 6-8. The synthesis method comprises: performing an enzyme catalytic reaction on crotonic acid in the presence of the high stereoselectivity amino-cleaving enzyme to obtain (S)-3-aminobutyric acid with a chiral purity e.e. of 99.5% or more; reacting (S)-3-aminobutyric acid with di-tert-butyl dicarbonate, and then reacting with ammonium bicarbonate, and further performing dehydration and deprotection to obtain (S)-3-aminobutyronitrile hydrochloride. The application combines chemical method and enzyme method, avoids the use of highly toxic substances, is safe and efficient, improves the economic benefits in the industrialization process, and the overall route reflects the value of green chemical synthesis.

The present application relates to a kind of mediated zymolytic enzyme of brown algae and application of double transcription unit recombinant bacteria of oxyprotein and brown algae, belong to genetic engineering technical field, the recombinant engineering bacteria is composed of two transcription units, one transcription unit is GAP-alpha-Vgb-AOX1TT, another transcription unit is AOX-alpha-102C300C-AOX1TT, two transcription units are expressed in tandem in pPICZ alphaA plasmid.The present application also provides the preparation method and application of the recombinant bacteria.The recombinant bacteria of the present application can improve the secretion expression efficiency of brown algae zymolytic enzyme, especially can be expressed in low-oxygen environment.

This invention discloses a method for preparing chondroitin A oligosaccharide ferrous sulfate with high enzymatic hydrolysis content, belonging to the field of bioengineering technology. The invention includes the following steps: softening animal cartilage by soaking in water; pulverizing the softened cartilage using microwaves; enzymatically hydrolyzing the pulverized cartilage with chondroitin sulfate lyase; separating the solid and liquid phases using a disc centrifuge; adsorbing chondroitin sulfate onto the filtrate using a resin column; eluting the resin column with an aqueous iron salt solution; and desalting, concentrating, sterilizing, and drying the eluent to obtain the finished chondroitin A oligosaccharide ferrous sulfate product, effectively preserving the natural structure, bioactivity, and physicochemical properties of chondroitin sulfate.