30 results about "Uridine diphosphate glucose" patented technology

The invention discloses a genetically engineered bacteria used for producing stevia glycosyltransferase UGT76G1; a UGT76G1 coding gene is inserted between the restriction enzyme cutting sites EcoRI and XhoI of a PYes2 carrier, so as to construct a recombinant plasmid, and then the recombinant plasmid is introduced to expression host Saccharomyces cerevisiae YPH499 to obtain the engineered bacteria; and the coding gene of the UGT76G1 is GenBank, No. GenBank: AY345974.1, and the gene sequence of the coding gene is named as UGT (Udp Glucuronyl Transferase). The invention also discloses a construction method of the genetically engineered bacteria and the application of the bacteria to the production of rebaudioside A. According to the invention, under the condition that expensive UDPG (Uridine Diphosphate Glucose) is not added, cheap carbon source glucose is used as a substrate, the metabolic pathway of UDPG in the yeast is regulated, and then the rebaudioside A is produced from St glycosides through whole cell catalysis.

The invention discloses biomarker compositions, and provides biomarker compositions for diagnosing cervicitis and a cervical cancer. The biomarker compositions are characterized by at least comprising one or a combination of uridine-5'-monophosphate disodium, histamine, 3'-O-methyl-guanosine and phenyllactic acid. The biomarker composition for diagnosing the cervicitis is characterized by comprising one or a combination of uridine-5'-monophosphate disodium, L-cysteine, isocitric acid, uridine diphosphate glucose, adenosine monophosphate, inosine 5'-monophosphate, L-pipecolic acid, N-acetyl putrescine and saccharose. The biomarker composition for diagnosing the cervical cancer is characterized by comprising one or a combination of histamine, phenyllactic acid, citraconic acid, L-2-aminoadipic acid, 3'-O-methyl-guanosine, beta-D-glucosamine, glycerin and cholesterol sulfate.

The invention discloses a method for generating rebaudioside M by catalyzing rebaudioside A via immobilized glycosyltransferases, which comprises the steps of: (1) crosslinking glycosyltransferase UGT1 and glycosyltransferase UGT2 on activated chitosan beads to obtain immobilized glycosyltransferases; (2) adding rebaudioside A into a container, adding uridine diphosphate glucose, and adding the immobilized glycosyltransferases, and catalyzing obtain rebaudioside M. The immobilized glycosyltransferases according to the invention are easy to separate, can be repeatedly used, have high thermal stability and high operation stability, and can, when compared with free enzymes, directly yield a higher amount of rebaudioside M by catalyzing rebaudioside A via the immobilized glycosyltransferases;the free enzymes can only be used once, while the immobilized glycosyltransferases can be reused; after the immobilized glycosyltransferases are reused 10 times, the activity of the immobilized glycosyltransferases is still more than 50% of that of the immobilized glycosyltransferase used for the first time. The method is simple and easy to implement. The process steps are reduced, the cost is reduced, and the production efficiency is improved.

The invention provides a preparation method of tea containing salidroside. The method comprises the following steps: spraying p-hydroxyphenylethyl alcohol to growing camellia sinensis leaves, and picking to obtain the tea containing salidroside. The method provided by the invention is simple in step; a synthetic precursor, namely p-hydroxyphenylethyl alcohol, of the salidroside is directly supplied from the external world; and uridine diphosphate glucose and glucosyltransferase contained in the tea are taken as raw materials, and the salidroside is synthesized in a tea body. The prepared tea containing the salidroside and application of the tea are further protected in the invention.

The invention discloses optically active (P)-bicyclol-9-O-beta-D-glucoside and (M)-bicyclol-9-O-beta-D-glucoside applicable to medicines and a preparation method and application thereof. According tothe invention, bicyclol and uridine diphosphate glucose disodium salt are catalyzed by glycosyltransferase to synthesize optically active (P)-bicyclol-9-O-beta-D-glucoside and (M)-bicyclol-9-O-beta-D-glucoside in one step. The bicyclol-9-O-beta-D-glucoside prepared according to the invention can be used for preventing and treating liver diseases.

The invention discloses a method for producing uridine diphosphate glucose and special engineering bacteria for the method. The method for producing the uridine diphosphate glucose comprises the following steps of by using uridine monophosphate and sucrose as raw materials, producing the uridine diphosphate glucose under the action of the engineering bacteria, wherein the engineering bacteria arerecombinant bacteria for expressing functional protein and are obtained by introducing a gene for encoding the functional protein into starting bacteria, and the functional protein comprises polyphosphokinase, uridine monophosphate kinase and sucrose synthase. The method is of great significance to industrial production of the uridine diphosphate glucose.

The invention relates to a method for preparing rebaudioside A by fermentation and catalysis of bacillus subtilis. The method is characterized by comprising the following steps of: 1, connecting a glycosyltransferase UGT76G1 gene onto a PBR322 plasmid vector, transferring the plasmid vector to a competent state of bacillus subtilis, inoculating the bacillus subtilis into an LB culture medium, andculturing at 30-37 DEG C for 20-24 h; 2, inoculating into a seed tank according to an inoculation amount of 1-10% volume ratio, culturing, introducing air, and culturing at 30-37 DEG C for 20-24 h; 3,inoculating into a fermentation tank according to an inoculation amount of 1-10% volume ratio, culturing, introducing air, controlling the pH to be 6-8, and culturing at 30-37 DEG C for 48-72 h; 4, carrying out filter pressing to obtain bacillus subtilis thalli, re-suspending, carrying out high-pressure homogeneous crushing, and carrying out filter pressing to obtain crude enzyme solution; and 5,mixing stevioside, uridine diphosphate glucose, phosphate buffer solution and the crude enzyme solution according to a mass ratio of (5-10): (1-5): (40-60): (5-20), and carrying out a reaction at 25-40 DEG C for 12-48 h. The method in the invention has the advantages that: the yield of the rebaudioside A is increased, and can reach 85.3 g/L; and the method is few in operation step, low in production cost and easy for industrial production.

The invention relates to the technical field of biology, in particular to a method for preparing uridine diphosphate glucose by means of an immobilized enzyme method. The method specifically comprisesthe steps of: (1) preparing UDPG (uridine diphosphate glucose) production enzymes; (2) immobilizing the UDPG production enzymes; and (3) separating a product. According to the invention, two UDPG production enzymes UGP and SPA are adopted, and the UDPG can be synthesized by just two-step enzymatic reaction; compared with a complex process for producing UDPG by chemical and fermentation methods, the method here has a simpler reaction process, with the reaction easier to control, and the product quality being more stable; the UDPG is prepared by means of an immobilized enzyme catalysis method,and the immobilized enzymes can be continuously and repeatedly used many times, so that the production cost is greatly reduced; in addition, a large amount of pigments, other types of nucleotides andother impurities introduced by the use of yeast are avoided, so that purification is easier; the method is suitable for large-scale production of UDPG.

The invention provides grifola frondosa UDP (User Datagram Protocol) glucosyltransferase as well as a coding gene and application thereof, and belongs to the field of edible mushroom molecular biotechnology and genetic engineering. According to the invention, UDP (User Datagram Protocol) glucosyltransferase UGT88A1 is obtained by cloning from Grifola frondosa and is named as GUFGT88A1, and the amino acid sequence of the UDP glucosyltransferase UGT88A1 is shown as SEQ ID NO. 2; the UDP glucosyltransferase is glucosyltransferase on which uridine diphosphate glucose (UDP-glucose) depends; the UDP glucosyltransferase can be used for catalyzing polysaccharide synthesis.

A recombinant microorganism for producing 2,3-butanediol consisting of selecting at least three groups from uridine diphosphate glucose phosphate uroglycan transferase gene (galU), acetyl alcohol dehydrogenase gene (acoA), acetyl phosphate transferase gene (pta), adenosine glucosylphosphate transferase gene (glgC), lactose dehydrogenase gene (ldhA), and phosphodiesterase gene (pdeC) which were modified.

The invention relates to a method for preparing rebaudioside D by fermentation catalysis of bacillus subtilis, which is characterized by comprising the following steps: (1) inoculating glycosyl transferase UGT76G1 and AtSUSY genes to a PBR322 plasmid vector, guiding into the bacillus subtilis, inoculating into an LB culture medium, and culturing for 20-24 hours at the temperature of 30-37 DEG C and the speed of 100-250 rpm; (2) inoculating into a seed tank according to the inoculation amount of 1%-10%, and culturing for 20-24 hours at the temperature of 30-37 DEG C at the speed of 200-400 rpm and the ventilation ratio of 0.1-1 V/V.min; (3) inoculating into a fermentation tank according to the volume ratio of 1%-10%, and culturing for 48-72 hours under the conditions that the speed is 200-1000 rpm, the ventilation ratio is 0.1-2 V/V.min, the temperature is 30-37 DEG C and the pH value is 6-8; (4) carrying out ultrafiltration on the fermentation liquor by adopting a ceramic membrane, resuspending bacteria, carrying out high-pressure homogeneous crushing, and carrying out ultrafiltration on the crushed liquor by adopting the ceramic membrane to obtain crude enzyme liquor; and (5) mixing stevioside, uridine diphosphate glucose, a Tris-HCl buffer solution and the crude enzyme solution according to a mass ratio of (5-10): (1-5): (40-60): (5-20), and reacting at 25-40 DEG C for 12-48 hours. The method has the advantages that two crude enzymes are obtained through one-time fermentation, the cost is low, and the conversion rate of rebaudioside D reaches 88-90 g/L.