35 results about "Hydroxylase gene" patented technology

The invention discloses a genetically engineered bacterium and an application of the genetically engineered bacterium in production of cis-3-hydroxy-L-proline. According to the genetically engineered bacterium, a proline dehydrogenase gene putA and a proline transporter gene putP in a starting bacterium are silenced, and meanwhile, a gamma-glutamyl kinase mutant gene proBD107N, a gamma-glutamyl phosphate reductase gene proA and a cis-3-hydroxy-L-proline hydroxylase gene P3H are subjected to overexpression. And by comparing a plurality of common escherichia coli, the cis-3-hydroxy-L-proline hydroxylase gene P3H and different gamma-glutamyl kinase mutant genes, the highest yield combination is preferably selected. The strain takes glucose as a raw material, so that the production cost is greatly saved, and the strain has a relatively good industrial utilization value.

The invention provides a genetically engineered bacterium for producing astaxanthin, and the genetically engineered bacterium takes Yarrowia lipolytica Po1f as an original strain and contains a GGS gene, an HMGR gene, a beta-carotene synthesis gene, a beta-carotene ketolase gene and a beta-carotene hydroxylase gene. The invention further provides the Yarrowia lipolytica MC247, and the preservation number of the Yarrowia lipolytica MC247 is CGMCC NO.29699. According to the Yarrowia lipolytica MC247, the high-yield astaxanthin is obtained, a new technical scheme is provided for efficient and green manufacturing of the astaxanthin, and industrial production of the astaxanthin is facilitated.

The present invention relates to the field of bioengineering, and in particular to an engineering bacterium with high yield of astaxanthin and its construction method and application. The present invention starts with wild-type Yarrowia lipolytica, introduces four genes of carB, carRP, crtZ and crBKT through a non-homologous recombination strategy, further overexpresses the key enzyme encoding gene in the mevalonate pathway (MVA) pathway to increase the yield of astaxanthin, and realizes the efficient production of the natural product astaxanthin. At the same time, the recombinant Yarrowia lipolytica provided by the present invention uses copper ion induction expression promoter MT-2 to control the expression of β-carotene ketolase crBKT gene, so that it keeps spatiotemporal differential expression with β-carotene hydroxylase gene crtZ, and when high concentration of intracellular zeaxanthin accumulates, copper ions are added to induce the expression of β-carotene ketolase crBKT gene, reduce the production of intermediate metabolic byproducts, and promote the efficient production of astaxanthin.

The present application is suitable for the field of molecular biology and biotechnology, and provides a maize gibberellin 13-hydroxylase gene ZmGA13ox1 and application thereof. The maize gibberellin 13-hydroxylase gene ZmGA13ox1 has a nucleotide sequence shown as SEQ ID NO:1 in the sequence listing. The maize gibberellin 13-hydroxylase protein is encoded by the maize gibberellin 13-hydroxylase gene ZmGA13ox1, and has an amino acid sequence shown as SEQ ID NO:2 in the sequence listing. The present application provides the nucleotide sequence of the ZmGA13ox1 gene in the maize inbred line B73 and the amino acid sequence information of the protein encoded thereby, and uses the agrobacterium-mediated transformation technology to introduce a plant expression vector containing the gene into Arabidopsis thaliana, and cultivates a homozygous T3 generation transgenic Arabidopsis thaliana plant. The experimental results show that the overexpression of the ZmGA13ox1 gene in Arabidopsis thaliana has a promoting effect on the root growth.

Gene-modified, lymphoid-tissue-homing dendritic cells that comprise a 1-alpha-hydroxylase gene and a retinaldehyde dehydrogenase 2 gene, where the 1-alpha-hydroxylase gene is expressed to produce functional 1-alpha-hydroxylase enzyme and the retinaldehyde dehydrogenase 2 gene is expressed to produce functional retinaldehyde dehydrogenase 2 gene enzyme. A method for treating one or more than one inflammation-related condition or disease, the method comprising administering gene-modified, lymphoid-tissue-homing dendritic cells that comprise a 1-alpha-hydroxylase gene and a retinaldehyde dehydrogenase 2 gene, where the 1-alpha-hydroxylase gene is expressed to produce functional 1-alpha-hydroxylase enzyme and the retinaldehyde dehydrogenase 2 gene is expressed to produce functional retinaldehyde dehydrogenase 2 gene enzyme.

The invention belongs to the technical field of biology, and particularly relates to an RNAi target gene for preventing and controlling sitobion avenae and design and application of dsRNA of the RNAi target gene. The sitobion avenae tyrosine hydroxylase gene is obtained through sequencing, annotation and verification, and the nucleotide sequence of the sitobion avenae tyrosine hydroxylase gene is SEQ ID No: 1; a sitobion avenae tyrosine hydroxylase gene is used as a target to design dsRNA, a primer with a T7 promoter sequence added at the 5'end is designed, and the nucleotide sequence of the obtained tyrosine hydroxylase dsRNA is SEQ ID No: 4. The sitobion avenae tyrosine hydroxylase gene is taken as a target gene, and sitobion avenae tyrosine hydroxylase dsRNA is correspondingly designed and is applied to prevention and control of sitobion avenae, so that the growth and development of the sitobion avenae are blocked, and a lethal effect is generated.

The invention discloses a genetically engineered bacterium for producing triacetyl sphingosine as well as a construction method and application of the genetically engineered bacterium, and relates to the technical field of biology. According to the present invention, the sphingosine C-4 hydroxylase gene in the Schiff Hammosaccharomyces cerevisiae is inactivated, such that the metabolic pathway of the Schiff Hammosaccharomyces cerevisiae is reconstructed, and the metabolic flow is efficiently guided to the synthesis of the triacetyl dihydrosphingosine, such that the large-scale production of the triacetyl dihydrosphingosine is stably achieved, and the significant industrial application potential and the significant economic benefit are represented.

The disclosure related to water-miscible forms of astaxanthin produced by transgenic red algae strains transformed to express β-carotene ketolase and hydroxylase genes from a green algae. These water-miscible forms of astaxanthin are aggregations of astaxanthin secreted by the transformed red algae. The particles accumulate in the extracellular space and are enriched in stationary phase of the algae culture, which enables simple and efficient isolation and purification of astaxanthin from algae. Thus, also disclosed herein are methods of isolating and / or purifying astaxanthin from transgenic algae comprising separating algae cells from its culture solution to collect the supernatant and then centrifuging the supernatant to pellet astaxanthin for collection and optionally further purification.

The present invention discloses a recombinant Escherichia coli and a method for synthesizing 5-hydroxyferulic acid from ferulic acid, belonging to the field of biotechnology. In the present invention, the 4-hydroxyphenylacetic acid-3-hydroxylase gene derived from Pseudomonas aeruginosa is heterologously expressed in Escherichia coli, and a recombinant bacterium capable of directly converting ferulic acid into 5-hydroxyferulic acid is successfully constructed. The whole-cell catalysis can produce 5-hydroxyferulic acid with a yield of up to 1.97 g / L and a productivity of 91% within 24 h. The recombinant bacterium and the whole-cell catalysis system provided by the present invention have simple processes and are environmentally friendly, laying a certain foundation for industrial production.

The invention relates to a fritillaria cirrhosa cholesterol 22 (R)-hydroxylase FcircCYP90B27 gene and application thereof, and belongs to the technical field of biology. The nucleotide sequence of the fritillaria cirrhosa cholesterol 22 (R)-hydroxylase FcircCYP90B27 gene is as shown in SEQ ID NO. 1, and the amino acid sequence of the encoded protein is as shown in SEQ ID NO. 2. The fritillaria cirrhosa cholesterol 22 (R)-hydroxylase FcircCYP90B27 gene disclosed by the invention can be used as a biosynthesis regulation gene of 22 (R)-hydroxycholesterol, is applied to preparation of 22 (R)-hydroxycholesterol, and is remarkable in application prospect and easy to popularize and apply.

The invention belongs to the field of gene engineering, and relates to a gene combination for synthesizing luteoloside, a recombinant plasmid combination, a strain and a method for heterologous biosynthesis of luteoloside. The gene combination comprises the following genes: a flavone hydroxylase gene TbFH; a flavone synthase II gene TbFNS-1 or TbFNS-2, and a flavone synthase II gene TbFNS-1 or TbFNS-2; the UDP glucosyltransferase gene is TbUGT-1, TbU75L1, U8089 or U16269, and the UDP glucosyltransferase gene is The invention provides a potential method for replacing traditional plant extraction or chemical synthesis, and lays a foundation for further transforming efficient biosynthesis through metabolic engineering.

The embodiment of the invention discloses recombinant XVII humanized collagen and a preparation method thereof, relates to the technical field of recombinant protein, and aims to solve the technical problem of low preparation efficiency of existing XVII type collagen. The preparation method of the recombinant XVII humanized collagen comprises the following steps: obtaining an amino acid sequence containing an integrin receptor site "RGD" and a binding site "KGHK"; based on the amino acid sequence, obtaining recombinant humanized collagen protein granules; the method comprises the following steps: obtaining recombinant lysine hydroxylase gene plasmids on the basis of a human gene LH2; and obtaining the recombinant XVII humanized collagen on the basis of the recombinant humanized collagen protein plasmid and the recombinant lysine hydroxylase gene plasmid.

The invention relates to the field of medicinal plant genetic engineering, and particularly discloses a CYP450 gene participating in lignan synthesis in a plant and a coding product thereof. The invention aims to analyze the oxidation modification step of the lignan biosynthetic pathway in the plant, and provides a gene element for the subsequent synthetic biological application. The invention discloses a C-5 hydroxylase gene sequence and an amino acid sequence thereof required by biosynthesis of aryl tetrahydronaphthalene lignans in podophyllum hexandrum for the first time. Reference is provided for subsequent rational design and directional transformation of lignan compound structure modification enzymes, development and utilization of biosynthetic pathway elements and synthetic biology heterologous efficient production of podophyllotoxin compounds.

The invention belongs to the field of gene engineering, and relates to a gene combination for synthesizing delphinidin, a recombinant plasmid combination, a strain and a method for heterologous biosynthesis of delphinidin. The gene combination comprises the following genes: a flavanone 3-hydroxylase gene TbF3H, a P450 cytochrome oxidase gene TbCYP, a flavanonol 4-reductase TbDFR, an anthocyanin synthetase TbANS and a P450 cytochrome reductase gene LjCPR, and the flavanone 3-hydroxylase gene TbF3H, the P450 cytochrome oxidase gene TbCYP, the flavanonol 4-reductase TbDFR, the anthocyanin synthetase TbANS and the P450 cytochrome reductase gene The invention provides a potential method for replacing traditional artificial cultivation or submerged fermentation acquisition, and lays a foundation for further transforming efficient biosynthesis through metabolic engineering.

Provided is a use of gene encoding gibberellin 3B-hydroxylase of G. max, GmGA3ox1. The use of gibberellin 3β-hydroxylase gene of G. max, GmGA30x1, set forth in SEQ ID NO:1, is in genetic engineering of seed weight of Arabidopsis thaliana and Glycine max. In A. thaliana, overexpression of GmGA3ox1 can complement the low seed weight phenotype of an atga3ox1 mutant. In G. max, overexpression of the excellent haplotype of the gene can significantly improve the seed weight of G. max.

The present invention provides a transgenic plant having enhanced stress resistance and / or with a change in flower color, preferably with a change in flower color to blue, or a self-fertilized or filial generation thereof, or a part of a plant body, a tissue or a cell thereof, and provides a method capable of producing 2-hydroxyflavanone in a simple and efficient manner. The present invention uses a polynucleotide encoding an enzyme belonging to the 2-ketoglutaric acid dependent dioxygenase (ODD) family and having an activity of hydroxylating position 2 of a flavanone.

The invention belongs to the technical field of gene engineering, and discloses a recombinant yeast strain for synthesizing astaxanthin as well as a construction method and application of the recombinant yeast strain. The recombinant plasmid provided by the invention comprises the following gene segments: a promoter TEF1p, a beta-carotene hydroxylase gene CrtZ, a cis-acting element IGG6, a beta-carotene ketolase gene CrtW and a terminator CYC1t which are spliced in sequence. A specific cis-acting element IGG6, a promoter, a terminator and exogenous CrtZ and CrtW are selected, a gene segment capable of producing astaxanthin is obtained, a recombinant plasmid is constructed from the gene segment, the recombinant plasmid is transformed into a saccharomyces cerevisiae strain, and high transformation of astaxanthin is achieved. Furthermore, the CrtZ and the CrtW are mutated by adopting error-prone PCR (Polymerase Chain Reaction) to obtain forward mutation, so that the conversion efficiency of the astaxanthin is further improved, and a more economic and more efficient way is provided for heterologous production of the astaxanthin by a microbial chassis.

The application belongs to the technical field of biology, and discloses a 7-amino-deacetylcephalosporin producing bacterium as well as a preparation method and application thereof. The 7-amino-deacetylcephalosporin producing bacterium is obtained by knocking out acetoxy synthase / hydroxylase genes and acetyltransferase genes in cephalosporium acremonium and introducing related genes for synthesizing 7-amino-3-deacetoxycephalosporin. The 7-amino-deacetylcephalosporin producing bacterium can be used to directly prepare high-yield and high-purity 7-amino-deacetylcephalosporin through microbial fermentation.

The invention discloses an eggplant flavonoid 3 ', 5'-hydroxylase gene and an application method thereof, and relates to the field of biochemistry, the gene sequence is shown as SEQ ID NO.1, and the amino acid sequence is shown as SEQ ID NO.2; the invention discloses a cloning method of the gene and a gene expression detection method. The gene is applied to creation of new eggplant varieties with different colors, creation of a rare blue system, creation of fruits rich in antioxidant delphinidin anthocyanin, production of natural delphinidin and production of delphinidin through fermentation engineering. According to the invention, on the basis of DNA high-fidelity polymerase, the CDS sequence of the eggplant SmF3 '5' H gene is cloned by using a PCR technology, Sanger sequencing verification is completed, and gene expression quantity detection is carried out. The eggplant SmF3 '5' H gene has huge application potential in the aspects of crop appearance and nutritional quality improvement.

This invention discloses a method for controlling the biotransformation of canlione by monitoring the expression level of hydroxylase gene. The steps are as follows: using *Aspergillus ochraceus* as the inoculum and canlione as the substrate, the expression level of the hydroxylase gene is monitored using real-time quantitative PCR during cell culture and transformation to determine the mid-logarithmic growth phase of the cell culture; canlione is then biotransformed again using *Aspergillus ochraceus* as the inoculum and canlione as the substrate, with an exogenous inducer added during the mid-logarithmic growth phase of the cell culture to improve the transformation rate. This method uses the hydroxylase gene expression level, a molecular biological indicator, as a key process parameter to guide the decision-making of substrate addition timing and the optimization of transformation conditions. It enables precise control of process optimization driven by gene expression data, achieving a significant increase in transformation rate and a significant reduction in transformation cycle, showing promising application prospects.

The invention relates to the technical field of microbial engineering, and discloses an L-hydroxyproline-producing de novo synthesis type corynebacterium glutamicum and a construction method thereof, and the corynebacterium glutamicum strengthens the biosynthesis flux from glucose to L-proline by overexpressing a key enzyme gene of an endogenous proline synthesis pathway; by introducing an exogenous proline-4-hydroxylase gene, the accumulated L-proline is efficiently converted into the L-hydroxyproline; meanwhile, by knocking out key genes of a competitive pathway, generation of by-products such as lysine and threonine is reduced, and metabolic interference is relieved. The invention further provides a construction method of the engineering bacterium. The construction method comprises the steps of vector construction, gene knockout and overexpression, transformation, screening and the like. According to the method, an economical and feasible foundation is laid for industrial biological manufacturing of the L-hydroxyproline, the overall conversion efficiency from a carbon source to a final product is improved, and the utilization rate of raw materials and the yield of the L-hydroxyproline are improved.

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 application discloses a kind of efficient conversion preparation hydroxyl tyrosol hydroxylase mutant and its application, belong to biotechnology field.The application provides a kind of hydroxylase mutant HpaB Y282H And HpaB S258C , using E.coli YMGR5A as host strain, with pEtac plasmid as carrier, expressed 4- hydroxyphenylacetic acid hydroxylase gene (GenBank accession number: CP053601.1) in escherichia coli and mutant HpaBC Y282H Or HpaB S258C ;The recombinant strain is fermented in 250mL flask with 50mL LB medium, and the yield of hydroxytyrosol is 1.82g / L, 1.81g / L respectively, and the yield of hydroxytyrosol is 49.98mM / L, 48.2mM / L respectively in the medium added with 50mM tyrosol, and the conversion rate is more than 99%, 96% respectively, higher than the achievement of most research teams at present.

The invention belongs to the technical field of biology, and discloses a saccharomyces cerevisiae engineering strain capable of efficiently producing dihydroquercetin, a mutant, a construction method and application. The mutant is obtained by performing point mutation on a flavonoid 3-hydroxylase gene CsF3H from citrus and a flavonoid 3 '-hydroxylase gene SmF3' H from silybum marianum which are subjected to heterologous expression, and the mutation sites of the point mutation are selected from any one, two or three sites of the 198th site, the 388th site and the 389th site. Finally, the yield of dihydroquercetin in a shake flask is increased by 15%-25% compared with that of a control strain (saccharomyces cerevisiae strain for overexpressing CsF3H and SmF3 'H). The engineering bacterium provided by the invention not only has the advantage of high yield, but also can be fermented through relatively simple method conditions, is easy to implement and control in industry and low in investment cost, and lays a foundation for efficient synthesis and further industrialization of dihydroquercetin.

The invention provides a flavanone C3-hydroxylase gene of camptotheca acuminate, a carrier, a protein and application, and belongs to the technical field of bioengineering.CYP71A hydroxylase genes CYP71AU223 and CYP71AU224 in the camptotheca acuminate are obtained through screening, plasmids are constructed and introduced into a saccharomyces cerevisiae WAT11 host cell, the host cell is used for inducing expression of two microsomal proteins of CYP71AU223 and CYP71AU224, and therefore the flavanone C3-hydroxylase gene of the camptotheca acuminate is obtained. The two microsomal proteins both show C-3 site hydroxylation activity on the key intermediate naringenin for flavone biosynthesis, and can be used for preparing the citrus aurantium. According to the invention, molecular docking and site-specific mutagenesis technologies are combined to determine substrate oriented stable key residues of two proteins CYP71AU223 and CYP71AU224, the catalytic activity of the key residues is improved, and the types of hydroxylase at C3 site of flavonoid compounds are enriched.

The spirosolane skeleton 23-position hydroxylase gene and the spirosolane skeleton 23-position acetyltransferase gene derived from S. chacoense, S. tuberosum, and S. lycopersicum are found to be involved with the biosynthesis of leptine, which achieves resistance against Colorado potato beetle.

The present invention discloses a bacterial tryptophan 5-hydroxylase and its application, belonging to the field of biotechnology. The amino acid sequence of the bacterial tryptophan 5-hydroxylase is shown in SEQ ID NO.1, SEQ ID NO.3 or SEQ ID NO.5, or has more than 50% homology and functional equivalence with the amino acid sequence shown in any one of them. The present invention identifies a new type of bacterial tryptophan 5-hydroxylase family that uses heme as a cofactor and can catalyze tryptophan to produce 5-hydroxytryptophan in vivo or in vitro. The enzyme can be used to catalyze tryptophan to prepare 5-hydroxytryptophan or to prepare melatonin. The present invention utilizes an engineered strain that heterologously expresses tryptophan 5-hydroxylase genes, methyltransferase genes and decarboxylase genes and lacks tryptophan dioxygenase genes as a fermentation strain, and specifically produces melatonin through a microbial fermentation method. The method is green, economical, efficient, and has industrial production prospects.

The invention discloses an application of a soybean flavanone-3-hydroxylase gene and a soybean flavanone-3-hydroxylase gene and a soybean flavanone-3-hydroxylase gene. The nucleotide sequences of the soybean GmF3H1, the soybean GmF3H2 and the soybean GmFNSII-1 are as follows: SEQ ID NO. 1, SEQ ID NO. 2 and SEQ ID NO. 3 respectively. The CRISPR / Cas9 is used for simultaneously knocking out GmF3H1, GmF3H2 and GmFNSII-1 in the soybean, so that the expression level of the CRISPR / Cas9 in the soybean can be obviously reduced. Compared with a wild type, the hundred-grain weight, the branching number and the protein content of the three-gene mutant material are also stably increased in multiple generations. Therefore, the soybean flavanone-3-hydroxylase genes GmF3H1 and GmF3H2 and the flavone synthase gene GmFNSII-1 disclosed by the invention can be applied to the aspects of soybean hundred-grain weight, plant type improvement and high-protein variety quality improvement of soybean knockout strains through gene engineering regulation.

The present application relates to a kind of fritillaria cirrhosa cholesterol 22 (R) -hydroxylase FcirCYP90B27 Gene and application, belong to the field of biotechnology.Fritillaria cirrhosa cholesterol 22 (R) -hydroxylase FcirCYP90B27 Gene nucleotide sequence as shown in SEQ ID NO.1, the amino acid sequence of encoding protein as shown in SEQ ID NO.2.The fritillaria cirrhosa cholesterol 22 (R) -hydroxylase FcirCYP90B27 Gene of the present application can be used as 22 (R) -hydroxylase biosynthesis control gene, and be applied to the preparation of 22 (R) -hydroxylase cholesterol, application prospect is remarkable, easy to popularization and application.

The invention provides a C24-site hydroxylase gene CYP7140E67 of stamen eugenin C, a CYP7140E67 protein and application of the C24-site hydroxylase gene CYP7140E67 and belongs to the technical field of genes. According to the invention, the key enzyme for catalyzing hydroxylation of the C24 site of the stamen eugenin C is successfully found and identified, the blank of research on the catalytic enzyme in aesculus chinensis is filled, and a key gene and a protein coding sequence are provided for biosynthetic pathway analysis and synthetic biology research of the original aescin and derivatives thereof. Moreover, the proaescin is heterogeneously synthesized by expressing the CYP714E67 in the Bensi tobacco leaves, a living body biosynthesis system is constructed, the CYP714E67 is heterogeneously expressed by utilizing saccharomyces cerevisiae, and the in-vitro synthesis of the proaescin is realized by virtue of yeast microsomes, so that a flexible path is provided for industrial production; the problem of resource supply of aescin A is solved, and a solid foundation is laid for promoting cultivation of new varieties of high-yield plants and industrial biosynthesis.