18 results about "Hydroxylase gene" patented technology

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 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.

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 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 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.

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.

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 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.

The invention relates to the field of gene engineering, and discloses a Golden-Gate assembly system, a recombinant strain containing the Golden-Gate assembly system, a construction method and application of the recombinant strain, and a method for producing stevioside. The assembly system comprises a connection sequence I, a selection marker gene, a connection sequence II, a promoter I, a connection sequence III, a kaurene oxidase gene, a connection sequence IV, a terminator I, a connection sequence V, a promoter II, a connection sequence VI, a kaurene acid 13-hydroxylase gene, a connection sequence VII, a terminator II and a connection sequence VIII which are sequentially connected. According to the Golden-Gate assembly system, synchronous directional assembly of stevioside synthesis pathways is achieved through the eight specific connection sequences, the assembly efficiency of the stevioside synthesis pathways is greatly improved, the conversion rate of the synthesis pathways is increased, then the yield of stevioside is increased, and the Golden-Gate assembly system has good industrial prospects.

The invention belongs to the field of plant tissue culture, and particularly relates to a method for promoting taxol accumulation of taxus chinensis cells by overexpressing hydroxylase. The invention provides a method for promoting accumulation of paclitaxel in taxus chinensis cells, which comprises the following steps: constructing taxus chinensis cells overexpressing taxane-5 alpha-hydroxylase genes, performing tissue culture amplification, and extracting paclitaxel from the amplified taxus chinensis cells, wherein the taxane-5 alpha-hydroxylase from the overexpressed Popus alba has the optimal effect, and the yield of paclitaxel can be increased by 8.44%.

This invention belongs to the field of biotechnology, specifically relating to the design and application of an RNAi target gene and its dsRNA for controlling the wheat aphid. The invention involves sequencing, annotation, and verification to obtain the wheat aphid tyrosine hydroxylase gene, whose nucleotide sequence is SEQ ID No: 1. Using the wheat aphid tyrosine hydroxylase gene as a target gene, dsRNA was designed, including the design of primers with a 5' end T7 promoter sequence, resulting in the nucleotide sequence of the tyrosine hydroxylase dsRNA, which is SEQ ID No: 4. This invention uses the wheat aphid tyrosine hydroxylase gene as a target gene and designs corresponding wheat aphid tyrosine hydroxylase dsRNA, both of which are applied to control the wheat aphid, leading to inhibited growth and development and a lethal effect.