14 results about "Lyase Gene" patented technology

The invention discloses recombinant escherichia coli with high yield of N-acetylneuraminic acid and application of the recombinant escherichia coli, and relates to the technical field of biological genetic engineering. The invention relates to a recombinant escherichia coli, which is characterized in that the escherichia coli is taken as a host, and free expression of an N-acetylmannosamine epimerase gene yihS from Streptomyces xiamenensis or an N-acetylmannosamine epimerase gene ce3 from Bacteroides polymorpha and an exogenous N-acetylneuraminic acid lyase gene nano A is carried out; and carrying out recombinant expression on N-acetyl hexosamine 1-kinase nahK, a UDP-N-acetyl glucosamine pyrophosphorylase gene glmU and a UDP-N-acetyl glucosamine-2-epimerase gene neuC in the other synthetic route of the ManNAc. According to the recombinant escherichia coli with high yield of N-acetylneuraminic acid, the yield of N-acetylneuraminic acid can reach 23.08 g / L under a shake flask fermentation condition; a two-stage batch feeding strategy is adopted, the yield of N-acetylneuraminic acid in a 5L fermentation tank reaches 71.25 g / L, the molar conversion rate of GlcNAc reaches up to 57.60%, and the method has the potential of industrial application.

The application provides a method for increasing the length of a main panicle of rice, the number of secondary branches of the main panicle, the number of grains on the main panicle of rice and / or improving the yield of rice, which comprises overexpressing a cystathionine beta-lyase gene OsCBL in rice by using a recombinant vector containing a CDS coding sequence of the cystathionine beta-lyase gene OsCBL as shown in SEQ ID NO: 2 to obtain a transgenic plant. The transgenic plant obtained by the method has an increased number of grains per panicle and an increased yield per plant compared with normal plants. The method can be used for treating rice plants to improve the yield of rice and provides a new gene resource and a new idea for breeding rice with increased yield.

The invention discloses a Rogowski eutrophy alkali-producing engineering strain for producing p-coumaric acid as well as a construction method and application of the Rogowski eutrophy alkali-producing engineering strain. The preparation method comprises the following steps: by taking a chemoautotrophic microorganism Ropriavidus alcaligenes H16 as an original strain, firstly, by knocking out a synthetic route of a strain poly-3-hydroxybutyrate (PHB), introducing a tyrosine ammonia lyase gene from Rhodotorula glutinis to construct a basic engineering strain capable of producing p-coumaric acid, and then, carrying out fermentation on the basic engineering strain to obtain the p-coumaric acid. Then, endogenous 3-deoxy-7-phosphoheptanone acid synthase gene aroG1 and chorismate synthase gene aroC are overexpressed, and the total copy is increased, so that the p-coumaric acid synthetic pathway flux of the engineering strain is enhanced; and introducing NAD (P) transhydrogenase gene pntAB from Escherichia coli to improve the supply of cofactors, so as to finally obtain the eutrophy alcaligenes Rosei engineering strain capable of synthesizing p-coumaric acid by using CO2.

The present application relates to a kind of high catalytic activity artificial synthesis of ulvan lyase A21 and application, belong to functional enzyme technical field, the amino acid sequence of the artificial synthesis of ulvan lyase A21 as shown in SEQ ID NO.1, the present application also provides the enzyme preparation comprising the artificial synthesis of ulvan lyase A21, and the recombination expression vector and engineering bacteria comprising the gene of the artificial synthesis of ulvan lyase A21 coding ulvan lyase A21.The present application also provides the application of the artificial synthesis of ulvan lyase A21 in the preparation of ulvan oligosaccharide of the cleavage of ulva lactuca.The artificial synthesis of ulvan lyase A21 compared with wild ulvan lyase has better stability, and its catalytic activity compared with wild enzyme JpPL40A is increased by nearly 4.5 times.

The application belongs to the technical field of biotechnology, and particularly relates to a Clostridium perfringens bacteriophage lytic enzyme produced by Bacillus subtilis, a preparation method and application thereof, an optimized Clostridium perfringens bacteriophage lytic enzyme coding nucleic acid sequence according to the codon bias of Bacillus subtilis, and an optimized Clostridium perfringens bacteriophage lytic enzyme coding nucleic acid sequence as shown in SEQ ID NO. 1. The preparation method of the Clostridium perfringens bacteriophage lytic enzyme is as follows: firstly, a recombinant expression vector containing a Clostridium perfringens bacteriophage lytic enzyme gene is constructed, and then the vector is transferred into a Bacillus subtilis competent cell to obtain a Bacillus subtilis strain recombinant engineering bacterium. After the recombinant strain is induced to express, a fermentation liquor with Clostridium perfringens bacteriophage lytic enzyme activity can be obtained. The fermentation liquor can specifically inhibit the growth of Clostridium perfringens, and provides a safe, efficient and orally-taken preparation for treating animal diseases caused by Clostridium perfringens.

The invention relates to the technical field of bioengineering, and discloses a construction method and application of recombinant Escherichia coli engineering bacteria with high yield of ergothioneine, and the method comprises the following steps: by taking Escherichia coli as a chassis, integrating a mycobacterium smegmatis egtABCDE gene cluster at an ldhA site; a repressor protein gene metJ and a cysteine lyase gene yhaM are knocked out, a methionine adenosine transferase gene metK and a serine acetyltransferase gene cysE are overexpressed by using a strong promoter, and mutant metA, thrA, serA and hisG genes for relieving feedback inhibition are introduced, so that metabolic inhibition of methionine, cysteine and histidine pathways is systematically relieved; the invention also establishes a matched fermentation process. According to the invention, endogenous precursor supply and exogenous synthesis pathways are synergistically enhanced, the precursor supply bottleneck is solved, the yield of ergothioneine is increased, and the method is suitable for industrial production.

The invention discloses a mucus type pseudomonas aeruginosa bacteriophage separation method based on recombinant alginate lyase, according to the separation method, mucus type pseudomonas aeruginosa bacteriophage is obtained based on recombinant alginate lyase separation, and the amino acid sequence of the recombinant alginate lyase is shown as SEQ ID NO.2. An alginate lyase gene algL in genomic DNA of a pseudomonas aeruginosa standard strain PAO1 is used as a target gene, a recombinant plasmid is constructed, induced expression is completed in escherichia coli, and the recombinant alginate lyase with high purity and high alginate degradation activity is obtained. Based on the recombinant alginate lyase, the bacteriophage which shows a high splitting effect on the mucus type pseudomonas aeruginosa is screened, the problem that the mucus type pseudomonas aeruginosa bacteriophage is difficult to separate is solved, and a technical basis is provided for development of drugs for treating infectious diseases caused by the mucus type pseudomonas aeruginosa.

The application discloses a phenylalanine ammonia-lyase gene RkPAL , which is separated from red yeast (Rhodotorula rubra) Rhodosporidium kratochvilovae , has a nucleotide sequence as shown in SEQ ID NO:1, and encodes an amino acid sequence as shown in SEQ ID NO:2. The gene is connected with a carrier and is transferred into red yeast cells, and experimental results show that RkPAL overexpression of the gene can promote the red yeast to synthesize carotenoids and oil; and the application provides a reference for large-scale commercial production of carotenoids and oil.

This invention relates to the field of biotechnology, and discloses an engineered *Yarrowia lipolytica* strain that produces p-coumaric acid using glucose as a substrate, its construction method, and its applications. Construction method: A tyrosine ammonia-lyase gene is integrated using a CRISPR / Cas9 localization and integration method. TAL In *Yarrowia lipophila* strains, the DAHP synthase gene was enhanced. ARO4 , DHS1 and AROG Overexpression of tyrosine synthase TYR Genes and histidine phosphotransferases HIS5 The protease gene was further integrated to synthesize exogenous phenylalanine deamination and hydroxylation pathway genes for p-coumaric acid, including phenylalanine ammonia-lyase gene, cinnamate hydroxylase gene, and P450 reductase gene. The p-coumaric acid produced by the engineered *Yarrowia lipolytica* strain of this invention can reach a maximum yield of 1.7 g / L in shake flasks, and a yield of 30 g / L in a 5 L fed-batch fermentation tank, demonstrating significant industrial application value.

The invention discloses a recombinant vector for expressing bacteriophage lyase and a construction method and application of the recombinant vector. The recombinant vector for expressing the bacteriophage lyase is based on a pichia pastoris expression system. The recombinant vector for expressing the bacteriophage lyase comprises a bacteriophage lyase gene, a multiple cloning site, a replication start site, a promoter, a terminator, an alpha-factor signal peptide and a resistance selection marker. According to the recombinant vector of the bacteriophage lyase provided by the invention, the lyase gene is recombined onto the vector, so that the expressed lyase can be used for developing a novel feed additive for preventing necrotic enteritis of poultry directly or after being prepared into a preparation, and a food preservative for controlling clostridium perfringens pollution in meat; or as a therapeutic drug for local or systemic infections.

This invention discloses a chemoautotrophic alkaloid-producing strain of *Alcaligenes rocheborella*, its construction method, and its application. The strain is based on the chemoautotrophic microorganism *Alcaligenes rocheborella* H16 (… Cupriavidus necator Using H16 as the starting strain, the poly-3-hydroxybutyrate (PHB) synthesis pathway of the strain was first knocked out, and then a strain from Rhodotorula glutinis (H16) was introduced. Rhodotorula glutinis ) tyrosine ammonia lyase gene tal A basic engineered strain capable of producing p-coumaric acid was constructed. Subsequently, the throughput of the p-coumaric acid synthesis pathway in the engineered strain was enhanced by overexpressing the endogenous 3-deoxy-7-phosphate heptanulate synthase gene aroG1 and the branching acid synthase gene aroC, and by increasing the tal copy number. Further enhancement was achieved by introducing strains from *Escherichia coli* (…). Escherichia coli By improving the supply of cofactors through the NAD(P) transhydrogenase gene pntAB, a strain of Alcaligenes roximatelye that can synthesize p-coumaric acid using CO2 was finally obtained.

This invention discloses a recombinant engineered bacterium producing ergothioneine, its construction method, and its application, belonging to the field of gene recombination fermentation technology. This invention uses *Escherichia coli* as the starting bacterium to recombinantly express histidine trimethylammonium cysteine ​​sulfoxide synthase. egt1 Gene, L-histidine methyltransferase egtD Gene, histidine trimethyl inner salt cysteine ​​sulfoxide lyase egt2 Gene, methionine adenosine transferase metK Gene, adenosine kinase adK Genes and adenine phosphoribosyltransferase apt The gene was modified, and further metabolic modification was used to increase the supply of precursor substances histidine, cysteine, and methionine. The resulting recombinant engineered ergothionein-producing bacteria achieved a yield of 2.54 g / L after 48 h of shake-flask culture, and a yield of 18 g / L after further expansion to a 5L fermenter. This method reduces energy consumption and costs while ensuring ergothionein production, making it suitable for practical application.

This invention proposes an alginate lyase, a compound enzyme preparation, and their applications, belonging to the field of enzyme engineering technology. It solves the technical problem that a single alginate lyase cannot efficiently prepare alginate oligosaccharides directly from brown algae. The alginate lyase of this invention is isolated from Shewanella. Shewanella sp. HD5 The original alginate lyase gene of *Shewanella* was obtained through heterologous expression. Shewanella sp. HD5 The alginate lyase of this invention was deposited on September 22, 2025, at the China Center for Type Culture Collection, Wuhan, Hubei Province, with accession number CCTCC NO: M20252078. This invention provides a high-yield lyase obtained through heterologous expression. It is combined with cellulase, pectinase, and papain to form a complex enzyme preparation that can be directly applied to the enzymatic hydrolysis of brown algae to obtain uniformly polymerized alginate oligosaccharides. Furthermore, the complex enzyme preparation exhibits higher extraction efficiency of alginate oligosaccharides than a single alginate lyase. The prepared alginate oligosaccharides can be used in food, feed, and pharmaceutical fields.