403 results about "Enzyme Gene" patented technology

The invention provides a uracil production strain and a construction method and application thereof.According to the strain, on an E.coli UR14 genome by means of a CRIPSR / Cas9 gene editing technology, firstly, psuG genes, preTA genes, rutA genes and upp genes are knocked out, so that decomposition of uracil is blocked; then, a uridine phosphorylase gene udp and a pyrimidine-5 '-nucleotide nucleotidase gene ppnN are subjected to overexpression, and synthesis and accumulation of uracil are synergistically enhanced; and finally, overexpression of the ribose phosphate mutase gene pgm further enhances the conversion of a by-product ribose phosphate 1-precursor 5-ribose phosphate 1-pyrophosphate and improves the carbon utilization rate, and the obtained strain has good genetic stability and high fermentation yield, can stably produce uracil, and has wide application prospects.

The invention discloses a molecular marker remarkably related to the amylose content of wheat and application of the molecular marker, and belongs to the technical field of wheat breeding. Three mutation sites of wheat starch branching enzyme gene SBEIIa on 2A, 2B and 2D chromosomes and two mutation sites of SBEIIb on 2A and 2B chromosomes are identified, a KASP molecular marker system closely linked with SBEIIa and / or SBEIIb is developed based on the SNP sites, the detection process is simple and convenient, the cost is low, the throughput is high, and the application prospect is wide. According to the method, the high-amylose wheat material which contains the mutant genotype and can be stably inherited can be accurately and efficiently screened out, the breeding process of wheat variety quality improvement can be accelerated, and the method has important significance on creation of the high-amylose wheat material or molecular assisted breeding of the high-amylose wheat.

The invention belongs to the technical field of genetic engineering breeding, and particularly relates to application and a method of a transcription factor CaBTF3 for regulating capsorubin synthesis. According to the capsicum transcription factor CaBTF3 and the coding gene thereof provided by the invention, the biosynthesis of capsorubin can be influenced by adjusting the expression level of a key enzyme gene in a capsorubin synthesis route and regulating and controlling the development and quantity of fruit chloroplast, the expression of the CaBTF3 gene in capsicum fruits is inhibited, the accumulation amount of capsorubin in the capsicum fruits can be reduced, and the yield of capsorubin is increased. The color of the pepper fruit is further influenced. By enhancing the expression level of the CaBTF3 gene, the content of capsorubin in capsicum fruits can be increased, the gene is applied to plant genetic engineering breeding, a new effective way is provided for creating capsicum varieties with high capsorubin content, and the gene plays an important role in improving the economic benefits of the capsicum industry and popularizing and applying the capsorubin.

The invention discloses a genetically engineered bacterium and an application of the genetically engineered bacterium in production of farnesene. The genetically engineered bacterium takes saccharomyces cerevisiae as a chassis strain; an exogenous beta-farnesene synthase gene is integrated, an ERG9 gene is knocked out, a promoter of a truncated HMG1 gene is replaced with an ANB1 promoter, an exogenous transhydrogenase gene is integrated and driven by a glucose sensitive promoter, beta-farnesene synthase is mutated and fused with a transmembrane structural domain of ABC transporter protein PDR10, and then a chimeric expression unit is constructed. According to the present invention, the chassis bacteria are modified by at least one of the steps of overexpression of phosphatidylcholine synthetase, knockout of sterol ester synthetase and introduction of exogenous alkyl glycerol monooxygenase, and the obtained engineering bacteria are combined with the optimized fermentation process so as to significantly improve the farnesene synthesis efficiency and the production economy; therefore, the genetically engineered bacterium and the fermentation method provided by the invention have good application prospects.

The invention discloses an ergothioneine-producing recombinant engineering bacterium as well as a construction method and application thereof, and belongs to the technical field of gene recombination fermentation. The method comprises the following steps: by taking escherichia coli as a starting bacterium, carrying out recombinant expression on a histidine trimethyl inner salt cysteine sulfoxide synthetase egt1 gene, an L-histidine methyltransferase egtD gene, a histidine trimethyl inner salt cysteine sulfoxide lyase egt2 gene, a methionine adenosine transferase metK gene, an adenylate kinase adK gene and an adenine phosphoribose transferase apt gene; on this basis, supply of precursor substances including histidine, cysteine and methionine is further improved through metabolic transformation, the yield of the finally obtained recombinant engineering bacterium for producing ergothioneine reaches 2.54 g / L after 48 h of shake-flask culture, the yield of a 5L fermentation tank reaches 18 g / L after further enlarged culture, the yield of ergothioneine is guaranteed while energy consumption and cost are reduced, and the method is suitable for industrial production. The method is suitable for practical popularization.

The invention discloses recombinant xanthomonas campestris with xanthan gum hydrolase displayed on the surface and a construction method and application of the recombinant xanthomonas campestris. The recombinant xanthomonas campestris for surface display of xanthan hydrolase is constructed by introducing a recombinant vector obtained by fusion expression of an ice crystal nuclein (INP) gene and a xanthan hydrolase gene into xanthomonas campestris. The gel yield of recombinant xanthomonas campestris fermentation is twice that of a wild strain and reaches 6.7 g / L. The strain can efficiently hydrolyze xanthan gum with high molecular weight (such as more than 100 wDa), the traditional production process which is relatively complicated is simplified, and a new method is provided for large-scale production of the xanthan gum with low molecular weight.

The invention relates to the technical field of gene engineering, in particular to an atractylodes macrocephala terpene synthase gene AmTPS9 and a product coded by the same and application of the gene AmTPS9. The gene AmTPS9 has one of the following nucleotide sequences: (1) a nucleotide sequence as shown in SEQ ID NO.1; and (2) a nucleotide sequence which is obtained by substituting, deleting or adding one or more nucleotides to the nucleotide sequence as shown in SEQ ID NO.1 and expresses protein with the same function. According to the invention, a coding gene of an atractylodes macrocephala terpene synthase gene (AmTPS) is cloned from the rhizome of the atractylodes macrocephala, and the synthase can be applied to a pathway prepared by taking farnesyl pyrophosphate (FPP) as a substrate. By utilizing the gene provided by the invention, the content of terpenoids in bighead atractylodes rhizome can be increased through a genetic engineering technology.

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 belongs to the technical field of plant genetic engineering, and particularly relates to a promoter of a pinus massoniana terpene synthase gene Pm TPS (-)-alpha-pinene and application of the promoter. According to the invention, a promoter fragment of a Pm TPS (-)-alpha-pinene gene and three homeopathic action elements with jasmonic acid response on the promoter fragment are obtained from pinus massoniana. A complete promoter fragment of the Pm TPS (-)-alpha-pinene gene of the pinus massoniana is transformed into arabidopsis thaliana by means of agrobacterium mediation, and the promoter can drive expression of a GUS gene in leaves of the arabidopsis thaliana or nicotiana benthamiana. GUS staining and GUS enzyme activity of transgenic Bensi tobacco leaves are reduced along with deletion of jasmonic acid response cis-acting elements on the promoter. Furthermore, the yield of pine resin terpenoids can be increased by utilizing the promoter and the homeopathic element thereof.

The invention discloses a recombinant bacterium capable of increasing the yield of alpha-bisabolol as well as a preparation method and application of the recombinant bacterium, and belongs to the technical field of microorganisms. The invention aims to improve the yield of alpha-bisabolol and enhance the tolerance of a host to an organic solvent. The invention provides a recombinant bacterium for improving the yield of alpha-bisabolol. Escherichia coli is used as a starting strain; the method comprises the following steps of: overexpressing an acetyl CoA acyltransferase / HMG-CoA reductase mvaE gene, an HMG-CoA synthetase mvaS gene, a 2-methyl citrate dehydratase prpD gene, a mevalonate kinase ERG12 gene, a mevalonate 5-phosphate kinase ERG8 gene, a mevalonate 5-diphosphate decarboxylase ERG19 gene and an isopentenyl diphosphate isomerase idi gene, so as to obtain a recombinant vector; the gene is obtained from an alpha-bisabolol synthase gene of artichoke, a farnesyl diphosphate synthase ispA gene and an alpha-bisabolol synthase CcBOS gene of artichoke. The industrial process of synthesizing alpha-bisabolol by a biological method is promoted.

The application provides a promoter capable of improving the transcription level of a glycine nucleotide transcarbamylase gene and application, the promoter is a mutation based on the sequence shown in SEQ ID NO: 1, and two promoters capable of enhancing the transcription level of the glycine nucleotide transcarbamylase gene and capable of improving the expression of an alpha-amylase amyE gene from Bacillus subtilis subsp. subtilis str. 168 in coryneform bacteria are obtained.

This invention relates to the field of enzyme engineering technology, and in particular to a highly efficient lactone hydrolase mutant for degrading zearalenone and its applications. This lactone hydrolase mutant uses the lactone hydrolase gene from Monosporascus sp. GIB2 as a template, and performs a site-directed mutation at amino acid position 134 to obtain L134A, L134V, L134I, and L134M mutants. Under optimal conditions of pH 9.0 and 60℃, these single-point mutants can efficiently degrade zearalenone (ZEN) within 3 minutes. The enzyme activity for degrading zearalenone is significantly improved compared to the original enzyme, with degradation activities increasing by 1.22 (L134A), 1.25 (L134V), 1.17 (L134I), and 1.14 (L134M), respectively, demonstrating significant applications and economic value in the feed industry.

The invention belongs to the technical field of genetic engineering and microbial engineering, and particularly relates to a method for improving the production level of cytidine and application. According to the recombinant escherichia coli, a glucose-6-phosphate dehydrogenase gene zwf and a 6-phosphogluconate dehydrogenase gene gnd are subjected to overexpression. Specifically, the fusion expression of a glucose-6-phosphate dehydrogenase gene zwf and a 6-phosphogluconate dehydrogenase gene gnd is carried out, and the fusion expression is carried out by adopting Linkers with different properties. During application, the recombinant escherichia coli can be used for producing the cytidine by fermentation, so that the production level of the cytidine is improved. According to the invention, recombinant Escherichia coli which is over-expressed and fused with a glucose-6-phosphate dehydrogenase gene zwf and a 6-phosphogluconate dehydrogenase gene gnd is constructed, and the recombinant Escherichia coli can be used for efficiently fermenting and producing cytidine.

This invention belongs to the field of genetic engineering technology, specifically relating to an engineered bacterium that efficiently synthesizes curcumin using ferulic acid as a precursor, its construction method, and its application. This invention enhances ferulic acid uptake and precursor supply capabilities from the source by knocking out the fdc and mcrC genes and overexpressing the tnaT and accD genes. Furthermore, it achieves high-level stable expression of exogenous key enzyme genes DCS and CURS through codon optimization. Combined with the synergistic effect of a two-stage fermentation regulation process, a stepwise increase in ferulic acid conversion rate is achieved. In a fermentation system with an initial ferulic acid concentration of 5 g / L, the conversion rate of the recombinant strain of this invention increased from 28.6% to 55.2% compared to the unoptimized strain, and further reached 80.2% after fermentation process optimization. Simultaneously, it effectively inhibits the formation of the byproduct 4-vinylguaiacol, significantly improving the utilization rate of ferulic acid and reducing the raw material cost of curcumin biosynthesis.

The application belongs to the field of enzyme genetic engineering, and particularly relates to application of a sesquiterpene synthase and a mutant thereof in synthesis of enantiomeric-drimenol compounds, wherein the sesquiterpene synthase is a sesquiterpene synthase from Chenopodium quinoa Willd., and includes CqDMDS1 or CqDMDS2; the mutant of the sesquiterpene synthase is a mutant CqDMDS1 in which an amino acid at position 443 of CqDMDS1 is mutated F443A / G The application realizes heterologous expression of the sesquiterpene synthase and the mutant thereof from Chenopodium quinoa Willd. in an Escherichia coli and a N. benthamiana expression system, catalyzes synthesis of (-)‑8,11-driman-diol or (+)‑drimenol. It is found through activity test that the sesquiterpene synthase from Chenopodium quinoa Willd. has a significant inhibitory effect on plant pathogenic fungi Phytophthora nicotianae, Phytophthora sojae and Fusarium graminearum. The sesquiterpene synthase and the mutant thereof and the biological preparation of the catalytic product have important significance for development of biological pesticides.

This invention relates to a cytochrome P450 oxidase STO1, its encoding gene, and its applications. This invention is the first to clone and identify a cytochrome P450 oxidase gene, STO1, from potato responsible for catalyzing the synthesis of spirogelothoraxone, and to determine the nucleotide sequence and translated amino acid sequence of this gene. This invention provides expression vectors, recombinant microorganisms, or transgenic cell lines containing this gene. This invention constructs an engineered Saccharomyces cerevisiae strain for producing spirogelothoraxone, in which the strong promoter ERG9 is replaced by the weak promoter HXT1 in the strain's genome, and the STO1, AtCPR, tHMG1, and STS genes are integrated. This invention also provides a method for constructing this engineered strain and a method for producing spirogelothoraxone using this engineered strain, which can be used for the preparation of spirogelothoraxone.

The invention discloses a method for constructing a pressed butterfly mycin glycoside gene engineering strain by using glycosyl transferase and application of the pressed butterfly mycin glycoside gene engineering strain. A strong promoter kasOp * is introduced to the upstream of a glucoside transferase gene pie28 of Streptomyces psammoticus SCSIO NS126, the nucleotide sequence of the strong promoter kasOp * is as shown in SEQ ID NO. 1, and the nucleotide sequence of the glucoside transferase pie28 is as shown in SEQ ID NO. 2. The invention also discloses a preparation method of the Streptomyces psammoticus SCSIO NS126, and the preparation method of the Streptomyces psammoticus SCSIO NS126 comprises the following steps of: introducing the strong promoter kasOp * into the upstream of the glucoside transferase gene pie28. The streptomyces gene engineering strain Streptomyces sp. SK-kas28 constructed by the invention can efficiently express the chalcone glycoside, is widely applied to fermentation production of the chalcone glycoside, and is used for research and development of new drugs for treating kidney diseases and the like.

The application provides a coenzyme Q10-producing genetically engineered bacterium and application thereof.

The application discloses a construction method and application of a genetically engineered corynebacterium glutamicum for synthesizing a bacteriochlorophyllide amino acid shinorine, and belongs to the field of genetic engineering and fermentation technology. The genetically engineered corynebacterium glutamicum is constructed by taking corynebacterium glutamicum ATCC 13032 as a host strain, knocking out a competitive pathway gene, integrating a key enzyme gene in multiple copies on a genome, knocking out an acid production pathway gene and a serine dehydrogenase gene, and expressing a gene cluster for synthesizing shinorine on a plasmid, so that the synthesis yield of shinorine in a microbial cell is improved. The strain can produce shinorine in a flask, and 1.12 g / L of shinorine can be accumulated in 72 hours. In fed-batch fermentation in a 50 L fermenter, the strain can be cultured to 8.6 g / L.

The present disclosure provides transgenic mice for use as mouse models, and methods related thereto. The transgenic mice described herein comprise a humanized mouse telomerase (hmTert) gene included in a background mouse germline that provides shortened telomeres capable of reproducing humans and various human disease states.

This invention discloses a *Bacillus laterosporus* strain for the stable production of nonribosomal cyclic lipolipeptide and its construction method. The M1A gene in the Bogorol biosynthetic gene cluster is knocked out using CRISPR-Cas9 technology to block the Bogorol pathway, which competes with Brevicidine for substrates. Subsequently, the pMCCas9-S48 plasmid, which integrates S8 and S41 sgRNAs and their homologous arms as repair templates, is used to simultaneously knock out S8 and S41 family peptidase genes, preventing the expression of extracellular degradation enzymes. This invention also provides the application of the above strain in the preparation of nonribosomal cyclic lipolipeptide of Brevicidine and a method for preparing the nonribosomal cyclic lipolipeptide. This strain, through reducing metabolic competition and preventing product degradation, exhibits rapid cell growth, stable production of Brevicidine, and significantly reduced degradation rate, resulting in a 66.13% increase in fermentation yield compared to the unmodified strain. This invention solves the problems of degradation and unstable yield in the production of Brevicidine, provides an effective technical approach for the industrial production of Brevicidine, and provides important technical support for the development and application of novel antimicrobial peptide drugs.

The invention discloses a black fungus AMY protein and a coding gene and application thereof, and belongs to the technical field of protein engineering. The invention provides a black fungus AMY protein as well as an encoding gene and application thereof in order to excavate a key enzyme gene of a starch sucrose metabolic pathway in black fungus, and discovers and verifies that the black fungus AMY protein and the encoding gene thereof play a key regulation and control role in inducing synthesis of black fungus alpha-amylase through transcriptome sequencing. A solid foundation is laid for deeply exploring the function of a key gene of a starch sucrose metabolic pathway.

The invention relates to Corynebacterium glutamicum with high indigo yield as well as a construction method and application thereof, and belongs to the technical field of gene recombination fermentation. The recombinant corynebacterium glutamicum can be used for overexpression of the following genes: a D-glucose-6-phosphate dehydrogenase gene zwf, a II type deoxyarabinose type heptulose phosphate synthase gene aroG and a flavin-containing monooxygenase gene CcFMO. The recombinant corynebacterium glutamicum can weaken the following genes: a pyruvate kinase gene pyk and a tryptophan synthase gene trpB. The corynebacterium glutamicum is used for overexpressing a zwf gene, an aroG gene and a CcFMO gene, weakening a pyk gene and a trpB gene and enhancing the expression quantity of an indole synthetic pathway related enzyme gene, so that the yield of the corynebacterium glutamicum at a shake flask stage reaches 619mg / L in a fermentation production process of indigo under the condition of not additionally adding tryptophan. The invention provides the recombinant corynebacterium glutamicum with high indigo yield, which is low in production cost, good in stability and high in synthesis efficiency, and is suitable for practical popularization and application.

The invention relates to an euonymus japonicus triterpenoid synthase EhypOSC3 gene and application of the euonymus japonicus triterpenoid synthase EhypOSC3 gene in preparation of cycloartenol, and belongs to the technical field of biology. The nucleotide sequence of the euonymus japonicus triterpenoid synthase EhypOSC3 gene is as shown in SEQ ID NO.1, and the total length of the sequence is 2280bp; the amino acid sequence of the encoded protein is shown as SEQ ID NO.2, and 759 amino acid residues are encoded. The euonymus japonicus triterpenoid synthase EhypOSC3 gene disclosed by the invention can be used as a biosynthesis regulation gene of cycloartenol, is applied to preparation of cycloartenol, and is remarkable in application prospect and easy to popularize and apply.

The application discloses a method for screening garlic fructan adversity response key enzyme genes and metabolites based on transcriptome and metabolome, and the method can better analyze the relationship between genes and metabolites by jointly analyzing the fructan-related differentially expressed genes measured by the transcriptome and the fructan-related metabolism substances with obvious metabolism changes measured by the metabolome, and constructing a gene-metabolite network interaction diagram.

The application discloses a method for producing deoxythymidine triphosphate by using a recombinant strain, and comprises the following steps: step 1: using a CRISPR / Cas9-mediated gene editing method to perform targeted modification on an E. coli genome, and the targeted modification comprises the following contents: a. knocking out a nucleoside triphosphate pyrophosphatase gene (mazG / yhdE) to block a path of degrading dTTP into dTMP; b. knocking out a nucleotide nucleosidase gene (ppnN) to block a path of degrading dTMP into thymine; and c. knocking out a nucleotidase gene (yfbR / yfdR / ushA / umpH / umpG / yjjG) to block a path of degrading dTMP into dT. The application provides a method for producing deoxythymidine triphosphate by using a recombinant strain, and the conversion rate of the method reaches 80% to 90%, and after ion exchange chromatography purification, the purity can reach more than 99%.

The invention discloses an application of a soybean 2-hydroxyisoflavanone dehydratase gene in improving the growth of intercropping corn. The gene HID1 can promote soybean to synthesize isoflavone, recruit bacillus and promote mycorrhizal fungi infection, so that the growth of intercropped corn is promoted, and the gene HID1 has important significance in illuminating the biological function of the GmHID1 gene in leguminous crop intercropping and regulating and controlling the synergistic growth promotion of plants, mycorrhizal fungi and bacillus in intercropping.

The invention discloses a method for efficiently preparing fixed-point biotinylation recombinant protein in escherichia coli, and belongs to the technical field of biology. According to the invention, a weakened expression regulatory sequence regulates and encodes a biotin ligase (BirA) gene expression cassette and a target protein gene expression cassette encoding a sequence carrying a biotin receptor peptide tag (AVI-Tag) to be jointly constructed on an escherichia coli expression vector, so that efficient fixed-point biotinylation of a target protein in an escherichia coli body is realized. The method has the advantages of high biotinylation efficiency, high specificity, simplicity and convenience in operation and the like, can be widely applied to the fields of separation and purification of proteins, protein interaction research and the like, and has important practical application value.

The invention provides bacillus licheniformis for high-yield protein glutaminase as well as a construction method and application of the bacillus licheniformis, a target strain is obtained by knocking out an alkaline protease gene aprE and extracellular protease genes bpr, epr, vpr or wpr and heterologously expressing the protein glutaminase gene, an EPE strain is optimal in expression, PG enzyme activity reaches 0.358 U / mL and is improved by 3.55 times compared with that of a wild type, and the bacillus licheniformis has the advantages that the bacillus licheniformis is high in yield and high in protein glutaminase yield. In addition, the growth condition of each mutant strain is not obviously abnormal, and the growth speed is obviously increased. It is fully proved that the expression level of the heterologous protein PG can be remarkably improved by targeted knockout of the extracellular protease gene of the bacillus licheniformis, and a theoretical basis is laid for construction of an efficient protein expression host. The invention not only provides a new scheme for high-efficiency expression of PG, but also provides important reference for expression optimization of heterologous proteins of other gram-positive bacteria, and shows a wide industrial application prospect.

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.