169 results about "Hydroxylase gene" patented technology

The invention discloses a method for producing eriodictyol by reforming escherichia coli in metabolic engineering, belonging to the field of metabolic engineering. The step of transforming naringenin into the eriodictyol is successfully realized through a gene engineering technology by carrying out fusion expression on two genes, namely a flavone 3' hydroxylase gene tF3'H with a membrane binding sequence cut and a P450 reductase gene tCPR, in escherichia coli. A tyrosine ammonia lyase gene TAL from R.glutinis, 4-coumarate:coenzyme A ligase gene 4CL from P.crispum, a chalcone synthase gene CHS from P.hybrida and a chalcone isomerase gene CHI from M.sativa are simultaneously expressed to realize that naringenin is directly produced through tyrosine, so that an engineered strain for directly producing eriodictyol through tyrosine is established. According to the method, in order to increase the output of eriodictyol, an acetokinase ackA gene contained in an escherichia coli genome is removed by being knocked, an acetyl coenzyme A synthetase gene acs and an acetyl coenzyme A carboxylase gene ACC are excessively expressed, and the output of eriodictyol can finally reach 106.7 mg/L.

The invention provides a method for producing 9-alpha-hydroxyandrostenedione by microbial fermentation. Recombinant Mycobacterium smegmatis is fermented to convert phytosterin to produce the 9-alpha-hydroxyandrostenedione. The method comprises the following steps: constructing a recombinant expression vector pMV261-ksh; constructing a recombinant strain; and carrying out culture propagation and fermentation conversion. Gene engineering means are utilized to obtain the gene engineering strain for overexpressing 3-sterone-9-alpha-hydroxylase gene (ksh); after the strain is fermented for 120 hours, the conversion rate of the phytosterin is up to 95% above; and the method has the characteristics of high yield, fewer byproducts, short fermentation time simple extraction, small pollution, low pressure for environmental protection and the like, and has huge advantages in industrial production.

The invention discloses engineering bacteria for producing trans-4-hydroxy-L-proline and a construction method and an application thereof. The construction method of the engineering bacteria provided by the invention includes the steps of A1) and A2): A1) introducing a L-proline-4-hydroxylase gene, a glutamate-5-kinase gene and a glutamate-5-semialdehyde dehydrogenase gene into a receptor cell; and A2) knocking an alpha-ketoglutaricdehydrogenase gene, an isocitratlyase gene or a proline dehydrogenase gene of the receptor cell out, or replacing a pyruvic oxidase gene of the receptor cell with an acetylcoenzyme A synthetase gene; and carrying out a reaction of the recombinant cell and a substrate to obtain the trans-4-hydroxy-L-proline. Experiments prove that the production method of the trans-4-hydroxy-L-proline can be used for production of the trans-4-hydroxy-L-proline.

Provided are transformed chrysanthemum plants having blue flower color, their self-fertilized progenies or cross-fertilized progenies thereof, a vegetative propagated plants thereof, and a part, a tissue or a cell of the plant body. Anthocyanin 3′,5′-O-glucosyltransferase gene (CtA3′5′GT) derived from Clitoria ternatea and flavonoid 3′,5′-hydroxylase gene derived from Campanula (CamF3′5′H) are coexpressed in chrysanthemum petals.

The invention discloses a biosynthetic gene cluster of paquete amide and an application thereof. The biosynthetic gene cluster of paquete amide comes from (Streptomyces pactum)SCSIO 02999, and the cluster comprises five genes including heterozygous polyketone/ nonribosomal peptide synthetases genes ptmA, FAD dependent redox enzyme genes ptmB1, phytoene dehydrogenase genes ptmB2, cyclase genes ptmC and hydroxylase genes ptmD. According to the biosynthetic gene cluster of paquete amide and the application thereof, information in genes and proteins related to biosynthesis of the paquete amide provides theoretical foundations and materials for conducting genetic modification on the biosynthesis of multi-ring tetramate macrocylic lactam family. By conducting genetic modifications on the biological synthetic genes, 6 new structures antineoplastic paquete amide compounds pactamide A-F are obtained, and thus effective compound entities are provided for research and development of antineoplastic drugs.

The invention discloses a gene related to flavone composite in radix scutellariae. The gene includes one or several in flavone synthetase gene with a sequence shown in SEQ ID No.1, flavone 6-site hydroxylase gene with a sequence shown in SEQ ID No.3 and flavone 8-site hydroxylase gene with a sequence shown in SEQ ID No.5. The invention further provides primer composite for amplifying the gene, protein encoded by the gene, a recombinant vector, host cell or transgenic cell line containing the gene and application thereof. The protein encoded by the gene disclosed by the invention can participate in synthesizing baicalein and wogonin and catalyzing synthesis and hydroxylation reaction of flavone matters of the similar structures, can provide very good basis for producing effective active matters, provides theoretical basis for producing baicalein, wogonin and aglycone of the baicalein and the wogonin in a large scale and establishes solid basis for industrially producing other related flavonoid compound.

The present invention provides methods for producing human ceramide in a yeast cell.

The methods of the present invention comprise:

• • 1) introducing the sphingoid Δ4-desaturase gene (DES1) by transformation of the yeast cell; and
  • 2) abolishing the expression of the yeast sphinganine C4-hydroxylase gene (SUR2) by transformation of the yeast cell.

The present invention belongs to the field of molecular biology and gene technology, and is especially the nucleotide coding sequence of flavoid-3', 5'-hydroxylase gene expressed in butterfly orchid, and its application in changing flower color. The present invention relates to the recombinant expression vector including the said gene, the transgenic plant, nucleotide primer sequence for obtaining the gene, the oligonucleotide sequence probe for detecting endogenous expression mode and method of analyzing and identifying the expression mode of the endogenous gene in butterfly orchid. Transforming the gene into petunia obtains transgenic plant with differentially expressed flavoid-3', 5'-hydroxylase gene and altered flower color.

The present invention relates generally to a genetic sequence encoding a polypeptide having flavonoid 3′,5′-hydroxylase (F3′5′H) activity and to the use of the genetic sequence and/or its corresponding polypeptide thereof inter alia to manipulate color in flowers or parts thereof or in other plant tissue. More particularly, the F3′5′H has the ability to modulate dihydrokaempferol (DHK) metabolism as well as the metabolism of other substrates such as dihydroquercetin (DHQ), naringenin and eriodictyol. Even more particularly, the present invention provides a genetic sequence encoding a polypeptide having F3′5′H activity when expressed in rose or gerbera or botanically related plants. The instant invention further relates to antisense and sense molecules or RNAi-inducing molecules corresponding to all or part of the subject genetic sequence or a transcript thereof. The present invention further relates to promoters which operate efficiently in plants such as rose, gerbera or botanically related plants.

The invention relates to a synthesizing method of 4-hydroxyisoleucine. According to the method, isoleucine hydroxylase genes are screened out, recombinant expression plasmid is constructed, and fusion protein having an amino acid deaminase transmembrane domain and isoleucine hydroxylase is expressed, so that isoleucine hydroxylase is anchored to the surface of cells; then a complete cell catalysis technology is utilized to replace a cell thawing and breaking technology, a cell freezing technology is omitted, and freezing processing equipment is not needed; meanwhile, product separation difficulty caused when inclusion of the cells is spread to reaction liquid after the cells are broken is avoided, the production procedure is simplified, the energy consumption, the material consumption and the production cost are lowered by a large margin, the conversion ratio of 4-hydroxyisoleucine produced through the method reaches up to 99.0% or above, and the synthesizing method is more beneficial to industrialized production.

The invention relates to a method for improving characters of gossypol in cotton, and a use thereof. In the invention, expression of a cadinene-8-hydroxylase gene CYP 706 B 1 in cotton seeds is down-regulated specifically thus synthesis of gossypol in cotton seeds is inhibited effectively without an influence on synthesis of gossypol in other tissues. Through the method, characters of gossypol ina cotton plant are improved and an integrated utilization value of cotton is increased greatly.

The invention provides a technology for obtaining 5-hydroxytryptophane by fermenting an engineering bacterial strain BL 21-DE. The engineering bacterial strain is prepared by utilizing a reported rabbit Oryctolagus cuniculus trptophan hydroxylase gene to build and reconstruct an expression vector and carrying out plasmid conversion. The bacterial strain is utilized for one-step fermentation process optimization; and under the condition of not purifying tryptophan hydroxylase, substrate tryptophan is added in the fermentation mature period to obtain the final product of 5-hydroxytryptophane. The invention has easily obtained raw material and low cost, conversion rate is more than 70%, and yield is more than 80%.

Genomic DNA and cDNA encoding GA 3β-hydroxylase were isolated from rice. When the expression of these genes was suppressed in rice plants, the plants became dwarfed compared with the wild type plants.

The invention relates to a method for converting indole to prepare indigotine by engineering strains formed by recombining phenol hydroxylase genes into colon bacillus, which belongs to the technical field of biological engineering. The method comprises the following steps: obtaining phenol hydroxylase gene full-length sequences with the total length of 5490bp and the Genbank registration number of FJ610336 in high-efficient phenol-degrading bacteria Arthrobacter sp.W1 with the GenBank registration number of EU339930; connecting the phenol hydroxylase gene full-length sequences to carrier plasmids (pET28a (+) Vector); and converting the phenol hydroxylase gene full-length sequences into host cell colon bacillus to construct phenol hydroxylase genetic engineering strains. The full-cell dry weight concentration of the phenol hydroxylase genetic engineering strains in a biological conversation reaction solution is between 1 and 2 g/L, the indole concentration is between 100 and 600 mg/L, the glucose concentration is between 0.5 and 2 mmol/L, the rotation speed of a shaking table is controlled between 50 to 200 r/min at a temperature between 20 and 40 DEG C, and the oscillating reaction lasts 2 to 14 hours. The biological conversation method of the invention has the advantages of mild reaction condition, high product concentration and short production period, and is suitable for industrial production.

The invention provides a rose characterized by comprising a flavone and delphinidin added by a genetic modification method. The flavone and delphinidin are typically produced by expression of a transferred flavone synthase gene and flavonoid 3′,5′-hydroxylase gene, respectively. The flavone synthase gene is, for example, a flavone synthase gene of the family Scrophulariaceae, and specifically it may be the flavone synthase gene of snapdragon of the family Scrophulariaceae, or the flavone synthase gene of torenia of the family Scrophulariaceae. The flavonoid 3′,5′-hydroxylase gene is, for example, the pansy (Viola×wittrockiana) flavonoid 3′,5′-hydroxylase gene.

The present invention relates to phenol hydroxylase gene and its use. The present invention provides metabolic encoding gene capable of converting phenol into catechol; constitutes HF conjugal transfer plasmid; and obtains genetic engineering strain capable of degradation to phenol or enlarging degradation range of substrate. When the phenol hydroxylase gene is transferred to immobile calcium acetate bacillus PHEA-2, the phenol degrading capacity of the strain will be raised. When the hydroxylase gene is transferred to toluene degrading bacillus, the bacillus strain will obtain phenol degrading capacity and enlarged substrate degrading range.