95 results about "UDP Glucose" patented technology

Disclosed is a novel enantiomeric synthesis cermamide-like inhibitors of UDP-glucose: N-acylsphingosine glucosyltransferase. Also disclosed are novel intermediates formed during the synthesis.

The present invention is directed to compositions and methods related to the synthesis and modification of uridine-5′-diphospho-sulfoquinovose (UDP-SQ). In particular, the methods of the present invention comprise the utilization of recombinant enzymes from Arabidopsis thaliana, UDP-glucose, and a sulfur donor to synthesize UDP-SQ, and the subsequent modification of UDP-SQ to form compounds including, but not limited to, 6-sulfo-α-D-quinovosyl diaclyglycerol (SQDG) and alkyl sulfoquinovoside. The compositions and methods of the invention provide a more simple, rapid means of synthesizing UDP-SQ, and the subsequent modification of UDP-SQ to compounds including, but not limited to, SQDG.

The invention concerns the use of duplex oligonucleotides about 25 to 30 base pairs to introduce site specific genetic alterations in plant cells. The oligonucleotides can be delivered by mechanical (biolistic) systems or by electrpoporation of plant protoplasts. Thereafter plants having the genetic alteration can be generated from the altered cells. In specific embodiments the invention concerns alteration in the gene that encode acid invertase, UDP-glucose pyrophosphorylase, polyphenol oxidase, O-methyl transferase, cinnamyl alcohol dehydrogenase, ACC synthase and ACC oxidase or etr-1 or a homolog of etr-1, and plants having isolated point mutations in such genes.

Disclosed is a novel enantiomeric synthesis ceramide-like inhibitors of UDP-glucose: N-acylsphingosine glucosyltransferase. Also disclosed are novel intermediates formed during the synthesis.

The invention discloses a method for preparing Rebaudioside E through an enzyme method. By means of the method, through tomato source UDP-glycosyl transferase and potato source sucrose synthetase, with stevioside as the raw material, the Rebaudioside E is produced through one-step glycosylation; meanwhile, by means of the fixed-point mutation technology, the UDP-glycosyl transferase is transformed, and the yield of the Rebaudioside E is further improved. By means of the method, no UDP-glucose or UDP needs to be added, the UDP-glucose is obtained as the raw material of glycosylation under the sucrose synthetase decomposition effect of the UDP in a crude extraction solution and the added sucrose, a double-enzyme circulating reaction system is established, and the stevioside is effectively catalyzed to generate the Rebaudioside E. The yield of the Rebaudioside E can reach 65% or above; through the fixed-point mutation technology in the later period, N358F beneficial mutants are obtained through screening, and the yield of the Rebaudioside E is improved through the catalysis and can reach 85% or above at most. Compared with an existing Rebaudioside E preparing method, the method is simple in processing step, low in cost and high in yield and has important application value.

The invention discloses a recombinant bacterium and application of the recombinant bacterium to the generation of rebaudioside D by catalyzing rebaudioside A. The recombinant bacterium contains a tomato-derived glycosyltransferase UGTSL2 gene and a potato-derived sucrose synthase StSUS1 gene; the tomato-derived glycosyltransferase UGTSL2 gene is cloned between NdeI and XhoI sites of pRSFDuet-1 to construct a recombinant plasmid pRSFDuet-SL2; then the potato-derived sucrose synthase StSUS1 gene is cloned between NcoI and EcoRI sites of the pRSFDuet-SL2 to construct a recombinant plasmid pRSFDuet-SL2-SUS1; the recombinant plasmid pRSFDuet-SL2-SUS1 is transferred into a host cell to obtain the recombinant bacterium. After the recombinant bacterium is subjected to induction expression, the recombinant bacterium is added into a reaction mixture to catalyze the rebaudioside A to generate the rebaudioside D; in reaction, crude enzyme liquid obtained by crushing the recombinant bacterium is utilized and separation and purification of an enzyme are avoided; lyophilized powder does not need to be produced; UDP (Uridine Diphosphate) or UDP-glucose and any cell penetration agent or other chemical reagents do not need to be added into a reaction solution, so that the recombinant bacterium has a better environment-friendly property. The yield of the rebaudioside D can reach 10.8g / L.

Nucleic acid sequences from potato that encode the enzyme UDP-glucose:solanidine glucosyltransferase (SGT2) are disclosed. Recombinant DNA molecules containing the sequences, and use thereof, in particular, use of the sequences and antisense constructs to inhibit the production of SGT2 and thereby reduce the level of the more human-toxic of the two predominant steroidal glycoalkaloids α-chaconine and / or increase the level of the more insect-toxic α-solanine in Solanaceous plants such as potato are described.

The invention discloses a recombinant bacterium and application of the recombinant bacterium to preparation of rebaudioside M2 by catalyzing rebaudioside A. The recombinant bacterium contains a tomato-based glycosyltransferase UGTSL2 gene and a potato-based sucrose synthase StSUS1 gene at the same time; the tomato-based glycosyltransferase UGTSL2 gene is cloned between NdeI and XhoI sites of pRSFDuet-1 and is constructed to obtain a recombinant plasmid pRSFDuet-SL2; then the potato-based sucrose synthase StSUS1 gene is cloned between NcoI and EcoRI sites of the pRSFDuet-SL2 and is constructed to obtain a recombinant plasmid pRSFDuet-SL2-SUS1; the recombinant plasmid pRSFDuet-SL2-SUS1 is transferred into a host cell to obtain the recombinant bacterium. After the recombinant bacterium is subjected to induced expression, crude enzyme liquid is taken and is added into a reaction mixture to catalyze the rebaudioside A to generate the rebaudioside M2; in a reaction process, the crude enzyme liquid obtained by crushing the recombinant bacterium is utilized and separation and purification of an enzyme are avoided; lyophilized powder is also not needed and substrates including rebaudioside D and UDP or UDP-glucose and any cell penetration agent or other chemical reagents do not need to be added, so that the environmental friendliness is better. The yield of the rebaudioside M2 reaches 11.09g / L.

The invention discloses a method for synthesizing 3-O-glucose-glycyrrhetinic acid with an enzymic method and belongs to the technical field of biological engineering. Glycosyltransferase gene Unigene14953, Unigene20323UGT73C11, UGT73C5 and UGT73C6 from plant licorice, barbarea vulgaris or Arabidopsis are cloned or chemically synthesized, escherichia coli is preferably selected as host bacteria, and genetically engineered bacteria containing glycosyl transferase is established; recombinant glycosyl transferase serves as glycyrrhetinic acid substrate and uridine diphosphate glucose, and 3-O-glucose-glycyrrhetinic acid can be synthesized efficiently in pH value of 5.0-11.0 and at the temperature of 25-55DEG C conditions.

Disclosed is a method of preparing a composition comprising a compound represented Structural Formula (I): The method comprises the step of reacting an aldehyde compound R10CHO with an isonitrile compound represented by Structural Formula (II):

The present invention discloses a panax japonicus var. major glycosyl transferase UGTPjm1 gene and an application thereof in preparing ginsenoside Ro. A nucleotide sequence of the panax japonicus var.major glycosyl transferase UGTPjm1 gene is represented by SEQ ID NO:1. Panax japonicus saponin IVa and a glycosyl donor UDP-glucose are used as raw materials and a glucuronic acid group at a C3 position of the panax japonicus saponin IVa is once further glycosylated to form the ginsenoside Ro catalyzed by panax japonicus var. major glycosyl transferase encoded by the panax japonicus var. major glycosyl transferase UGTPjm1 gene. The biosynthesis regulation gene UGTPjm1 of the ginsenoside Ro is first identified and successfully verified. By synthesizing the ginsenoside Ro in vitro, management is more convenient, controllability is strong, demand for raw material planting can be reduced, agricultural land is saved, a production product is single, and separation and purification of the ginsenoside Ro in a late stage are facilitated. The panax japonicus var. major glycosyl transferase UGTPjm1 gene is used as a key gene for the biosynthesis of the ginsenoside Ro and can also be used for breeding studies of panax japonicus var. major and other plants.

The invention discloses a biosynthesis method of uridine diphosphate glucose and uridine diphosphate glucuronic acid. The method includes adopting soluble starch as a main initial raw material, conducting recombinant expression on high-temperature alpha-glucan phosphorylase and high-temperature sugar-1-nucloside phosphorylase in escherichia coli respectively, and utilizing high-temperature whole cell catalysis of expressed bacteria to synthesize uridine diphosphate glucose; on this basis, conducting recombinant expression on high temperature uridine diphosphate glucose dehydrogenase in the escherichia coli, coupling a synthesis system of the uridine diphosphate glucose to conduct high temperature whole cell catalysis to synthesize the uridine diphosphate glucuronic acid, and introducing awhole cell catalysis system of high temperature NADH oxidase into the synthesis system of the uridine diphosphate glucuronic acid to form a high temperature NAD+ / NADH circulating system to reduce theuse of coenzyme NAD+. The high temperature whole cell catalysis method is utilized to successfully avoid the interference of various metabolic pathways of the bacteria in the synthesis process and reduce the purification difficulty.

The invention provides application of arabidopsis thaliana UDP-glucose 4-isomerase gene At-UGE2, an overexpression mutant strain 35S:UGE2 and a deletion mutant strain uge2 in adjustment of plant yield correlated characters, adversity resistance characters and the like. The function of biological synthesis and conversion of the UDP-glucose and UDP-galactose, which are catalyzed by UDP-glucose 4-isomerase, in adjustment of plant growth and development as well as adversity stress response is disclosed, a high-quality gene is provided for plant variety improvement, a new method and a new mechanism for improving plant yield characters and adversity resistance abilities are explained, and important economic significance and application value are achieved.

The invention discloses an improvement of Astragaloside content by endogenous gene overexpression technology, wherein the two metabolism reaction coupled key enzyme genes of the polysaccharide metabolic pathway, i.e., ugp gene (uridine diphosphate glucose phosphorylase gene) and gbss gene (adenosine diphosphate glucose glycoside transferase gene) are modified in vitro, so as to reinforce the enhanced expression regulation element and construct intermediate carrier, and electrical perforation method is employed to transfer the expression intermediate carrier pBI-UG into Agrobacterium rhizogenes LBA-9402, thus obtaining polysaccharide synthesize double gene Radix Astragali hairyroot.