576results about "Glycosyltransferases" patented technology

The invention provides a uracil production strain and a construction method and application thereof.According to the strain, on an E.coli UR14 genome by means of a CRIPSR / Cas9 gene editing technology, firstly, psuG genes, preTA genes, rutA genes and upp genes are knocked out, so that decomposition of uracil is blocked; then, a uridine phosphorylase gene udp and a pyrimidine-5 '-nucleotide nucleotidase gene ppnN are subjected to overexpression, and synthesis and accumulation of uracil are synergistically enhanced; and finally, overexpression of the ribose phosphate mutase gene pgm further enhances the conversion of a by-product ribose phosphate 1-precursor 5-ribose phosphate 1-pyrophosphate and improves the carbon utilization rate, and the obtained strain has good genetic stability and high fermentation yield, can stably produce uracil, and has wide application prospects.

The invention relates to the technical field of synthetic biology and microbial engineering, in particular to recombinant escherichia coli for de novo synthesis of forsythiaside A. The invention provides recombinant escherichia coli for de novo synthesis of forsythiaside A. A preparation method of the recombinant escherichia coli comprises a step of improving an escherichia coli starting strain, and the escherichia coli starting strain is escherichia coli MG1655. In the fermentation process of the recombinant escherichia coli, effective accumulation of forsythiaside A in fermentation liquor can be achieved, and a foundation can be laid for subsequent construction of high-yield forsythiaside A engineering bacteria.

The invention discloses a Pseudomonas putida high-yield rhamnolipid fermentation medium based on a step-by-step optimization strategy, an optimization method and application thereof, through the step-by-step strategy of component screening-single-factor optimization-multi-factor interactive optimization, a mixed carbon source and a mixed nitrogen source are taken as cores, and a Plackett-Burman test, a steepest climbing test and Box-Behnken response surface design are combined, so that the high-yield rhamnolipid fermentation medium for the pseudomonas putida high-yield rhamnolipid is obtained. Optimizing to obtain a special fermentation culture medium; two genetic engineering strains estA-2 and estA-3 are constructed by heterologous expression of key genes, and the supply and extracellular transport efficiency of a rhamnolipid synthesis precursor is enhanced; the recombinant strain is applied to the optimized culture medium, and efficient production of a shake flask and a 5L fermentation tank is achieved. The rhamnolipid yield of the estA-3 strain in a 5L fermentation tank reaches 35.59 + / -2.18 g / L, the conversion rate is 0.282 g / g, the productivity is 0.404 g / L / h, the rhamnolipid yield, the conversion rate and the productivity are obviously superior to those in the prior art, and a key technical support is provided for industrial production of rhamnolipid.

The invention belongs to the technical field of gene engineering, and particularly relates to application of a TaMGD3 gene in improvement of plant starch quality and cultivation of a high-starch transgenic plant, including application of the TaMGD3 gene in promotion of wheat grain starch synthesis, application of the TaMGD3 gene in cultivation of the high-starch and / or high-amylopectin transgenic plant, and application of the TaMGD3 gene in improvement of plant starch quality and cultivation of the high-starch and / or high-amylopectin transgenic plant. The invention also discloses an application of the TaMGD3 gene in improvement of plant starch characteristics. A nucleotide sequence of the TaMGD3 gene is shown as SEQ ID NO. 1. The invention also provides a cultivation method of transgenic wheat. In wheat cultivation, wheat amylopectin quality improvement is realized through overexpression of the gene shown in SEQ ID NO.1. The TaMGD3 gene can improve the starch content of wheat grains, influence the particle size distribution of the wheat grain starch, improve the amylopectin proportion, improve the peak viscosity and the relative crystallinity of the starch, and facilitate the improvement of the cooking quality related to the wheat starch.

[Summary] [Problems] To provide a novel technique for improving the taste of a glycosylated steviol glycoside composition. [Solution] The present technique provides a method for producing a glycosylated steviol glycoside composition and a method for improving the taste of a glycosylated steviol glycoside composition, which include a step of treating a stevia extract and dextrin with cyclodextrin glucanotransferase derived from a microorganism belonging to the genus Anoxybacillus. The production method and taste improvement method according to the present technique may further include a step of treating with a carbohydrate-processing enzyme other than the cyclodextrin glucanotransferase derived from a microorganism belonging to the genus Anoxybacillus.

The invention relates to an engineered glycosyltransferase and a steviol glycoside glucosylation method. The present invention provides engineered glycosyltransferases (GTs), polypeptides having GT activity and polynucleotides encoding the enzymes, as well as vectors and host cells comprising the polynucleotides and polypeptides. The present invention provides engineered sucrose synthases (SuS), polypeptides having SuS activity and polynucleotides encoding the enzymes, as well as vectors and host cells comprising the polynucleotides and polypeptides. The invention also provides compositions comprising the GT enzymes and methods of using the engineered GT enzymes to prepare products having beta-glucose ligation. The present invention also provides compositions and methods for producing rebaudioside (e.g., rebaudioside M, rebaudioside A, rebaudioside I, and rebaudioside D). The invention also provides compositions comprising the SuS enzymes and methods of using the same. Methods for producing a GT enzyme and a SuS enzyme are also provided.

The invention belongs to the technical field of bioengineering, and discloses a high-yield hyaluronic acid bacillus subtilis strain for expressing a hyaluronic acid synthase gene, a construction method and application, and the construction method comprises the following steps: inserting a codon-optimized sphaasA gene into a lytH site in a bacillus subtilis 168 strain in a traceless manner by utilizing a CRISPR / dCas9 system; the preparation method comprises the following steps: synthesizing hyaluronic acid oligosaccharides, simultaneously integrating genes for expressing a synthetic pathway of hyaluronic acid oligosaccharides, reducing the transcriptional level of zwf and pfkA genes, weakening the metabolic flux of a competitive pathway, and finally increasing the expression of sphaasA and tuaD genes according to a ratio of 2: 1 by using plasmids and the interaction of protein scaffolds RIAD and RIDD. When the strain is fermented for 48 hours in a 5L fermentation tank, the yield of the hyaluronic acid reaches 38.02 g / L, and the average molecular weight HA Mw of the low-molecular-weight hyaluronic acid is about (5-13) * 10 < 4 > Da.

The invention provides a method for cultivating anti-aging glutinous highland barley, highland barley starch as well as a preparation method and application of the highland barley starch, and particularly relates to a method for improving the starch quality character of an existing high-quality glutinous highland barley variety, in particular to a method for cultivating anti-aging glutinous highland barley by utilizing a gene editing technology without exogenous DNA (Deoxyribose Nucleic Acid) integration. A method for fixed-point knockout of an SSIIa gene to cultivate an anti-aging glutinous highland barley new strain under the background of a "Zhengnongglutinous 232" variety comprises the following steps: providing an acceptor material derived from a highland barley variety "Zhengnongglutinous 232" with a glutinous character, and deleting a Wx gene function of the "Zhengnongglutinous 232"; the method comprises the following steps: assembling guide RNA (Ribonucleic Acid) designed aiming at an SSIIa gene and Cas9 protein in vitro to form a ribonucleoprotein RNP complex; introducing the RNP complex into cells of an acceptor material so as to enable Cas9 protein to generate double-chain breakage at a target site of the SSIIa gene; carrying out culture and plant regeneration on the introduced cells to obtain a T0-generation plant of which the SSIIa gene is edited; and screening plants with the function loss of the SSIIa gene, wherein the plants keep the glutinous character of adult agricultural glutinous 232.

The present invention relates to a method for hydrolyzing carbohydrates present in raw materials containing carbohydrates, comprising the step a) of contacting the raw material containing carbohydrate with a medium in which bacteria capable of secreting carbohydrate-hydrolyzing enzymes have been fermented, to a medium in which bacteria capable of secreting carbohydrate-hydrolyzing enzymes have been fermented, to a method for simultaneously liquifying and saccharifying starch, a method for simultaneously liquifying, saccharifying and fermenting starch, and to a method for producing a biomass, the method comprising the step of cultivating bacteria secreting carbohydrate-hydrolyzing enzymes on a medium comprising complex carbohydrates as a major carbon source.

The present application relates to methods for preparing rebaudioside D and rebaudioside M by reaction of uridine diphosphate (UDP)-glycosyltransferases; and compositions for preparing rebaudioside D and rebaudioside M, comprising uridine diphosphate (UDP)-glycosyltransferases.

The invention discloses a genetically engineered bacterium for biosynthesis of gastrodin as well as a construction method and application of the genetically engineered bacterium. According to the genetically engineered bacterium, through overexpression of sucrose permease CscB derived from escherichia coli W, sucrose phosphorylase Basp of bifidobacterium adolescentis, uridine triphosphoryl-glucose-1-phosphate uridine acyl transferase UgpA of bifidobacterium bifidum and UDP-glycosyl transferase RsUGT of hedyotis diffusa, an exogenous path for synthesizing UDP-glucose from sucrose is constructed in a cell, and the UDP-glucose is synthesized into UDP-glucose. And the glycosylation capability of the gastrodin precursor on hydroxybenzyl alcohol is enhanced. The invention also provides a construction method of the genetically engineered bacterium and a method for synthesizing gastrodin through whole-cell catalysis by using the strain and taking p-hydroxybenzyl alcohol and cane sugar as substrates. The genetically engineered bacterium is high in gastrodin synthesis efficiency, the molar conversion rate can reach 97.8%, the highest yield reaches 3.61 g / L, and a new green manufacturing scheme is provided for solving the problems that a traditional gastrodin production method is low in efficiency, high in cost, large in pollution and the like.

In various embodiments, provided herein are host cells, methods, and pharmaceutical compositions comprising gastrodin, wherein the gastrodin is produced by a genetically modified plant or plant cell, fungal cell, yeast cell, insect cell, or bacterial cell. In certain embodiments, the present disclosure provides methods and compositions for the production of gastrodin. In yet other embodiments, the present disclosure provides enhanced cells and methods for producing gastrodin.

The present invention provides a polypeptide that catalyzes the hydrolysis of lactose β-(1-4) glycoside linkages for converting lactose and N-acetylglucosamine (GlcNAc) into a galactooligosaccharide (GOS) composition rich in N-acetyllactosamine (LacNAc). [Solution] A functional recombinant β-hexosyl-transferase (rBHT) polypeptide is provided, which has at least 90% sequence identity with a specific sequence and includes an N-terminal shortening of at least one amino acid with respect to the specific sequence.

Provided are compounds, methods, and pharmaceutical compositions for reducing the amount or activity of GYSI RNA in a cell or subject, and in certain instances reducing the amount of GYSI protein in a cell or subject. Such compounds, methods, and pharmaceutical compositions are useful to ameliorate at least one symptom or hallmark of a glycogen storage disease. Such glycogen storage diseases include Lafora disease, adult polyglucosan body disease (APBD), Andersen's disease, and Pompe disease.

Provided is Thermothelomyces heterothallica (formerly Myceliophthora thermophila) genetically modified to produce glycoproteins having N-glycans of mammalian proteins, particularly human, companion animal, and other animal proteins, comprising a deletion or disruption of the alg3 gene, expression of an ER-targeted mannosidase 1 (alpha-1,2-mannosidase), and expression of an ER-targeted glucosidase 2 alpha-subunit. The Thermothelomyces heterothallica may also further comprise a heterologous GlcNAc transferase 1 (GNT1), a GlcNAc transferase 2 (GNT2), an STT3 subunit of a heterologous oligosaccharyl transferase, and a galactosyltransferase.

Disclosed herein are methods of producing a food product or food precursor. These methods can comprise, for example, (a) providing milk, (b) adding sucrose to the milk to form sweetened milk, (c) contacting the sweetened milk with at least one glucosyltransferase to form insoluble alpha-glucan in the milk, (d) acidifying the milk product to a pH below about 5.0, and (e) inactivating the glucosyltransferase by subjecting the acidified milk product to a temperature of about 70-110 °C. Such methodology can produce an acidified milk product having a stable sucrose level and typically also a stable level of texture. Such methodology can optionally use other types of food products / precursors instead of milk, as appropriate. One or more glucosyltransferases used in the methodology can be selected from (i) a glucosyltransferase that synthesizes alpha-1,6-glucan, and / or (ii) a glucosyltransferase that synthesizes alpha-1,3-glucan. Food products / precursors produced by this methodology are also disclosed.

The invention discloses application of a PRTFDC1 gene in promoting porcine skeletal muscle satellite cell differentiation, myotube formation and skeletal muscle development. According to the invention, overexpression and knock-down experiments of the PRTFDC1 are respectively carried out in pig MuSCs, and immunofluorescence, qRT-PCR and Western blot analysis are carried out on cells subjected to induced differentiation, so that the PRTFDC1 is effectively shown to play a positive regulation role in a pig MuSCs differentiation process, and a new marker gene which can be applied to pig molecular breeding and growth performance evaluation is provided; and a novel molecular target and a novel detection tool are provided for muscle development regulation and biological breeding.

The present disclosure relates to toxin-derived delivery constructs, in particular isolated non-naturally occurring delivery constructs, for oral delivery, comprising a bacterial toxin-derived delivery construct coupled to a biologically active therapeutic cargo; wherein the delivery construct is capable of delivering a bioactive cargo through epithelial cells via endocytotic transport; and wherein the delivery construct does not comprise a bacterial toxin-derived translocation domain or a bacterial toxin-derived catalytic (cytotoxic) domain.

A glucosyltransferase catalyzing RA to produce RM and a use thereof are provided. The glucosyltransferases derived from Cornus florida and Juglans regia L. are screened and obtained. It was found that the glucosyltransferase derived from the Cornus florida exhibited a relatively high enzymatic activity in catalyzing the RA to produce the RD, while the glucosyltransferase derived from the Juglans regia L. exhibited a relatively high enzymatic activity in catalyzing the RD to produce the RM. When these two enzymes were used together in catalyzing the RA to produce the RM, a relatively high conversion rate was achieved. The use of the glucosyltransferases derived from the Cornus florida and the Juglans regia L. for preparing the RM of SGs, has not been reported yet. The glucosyltransferase provides an excellent candidate protein resource and its corresponding gene sequence for the enzymatic conversion process of the SGs.

The present invention provides a method for producing immobilized cells for producing mannose, and a method for producing mannose, which include the steps of obtaining fermentation liquids of Escherichia coli or Bacillus subtilis in which α-glucan phosphorylase, phosphoglucomutase, glucose phosphate isomerase, mannose 6-phosphate isomerase, and mannose 6-phosphate phosphatase have been expressed by fermentation, and mixing the fermentation liquids to obtain a fermentation mixture.

The present invention relates to a method for producing a stable hydrogel-cell composition comprising the steps of:a) providing a reaction mixture comprising at least one polymerizable substrate and at least one enzyme capable of polymerizing said at least one substrate, and cells, andb) incubating the mixture of step a) to form a stable hydrogel-cell composition.

This invention provides modified proteins, immunogenic compositions, and vaccines containing modified proteins, their manufacture, and the use of such compositions in pharmaceuticals. [Solution] A modified EPA (Exotoxin A of Pseudomonas aeruginosa) protein containing a specific amino acid sequence is provided, which is modified in that it contains one (or more) consensus sequences selected from three specific amino acid sequences, and can be used as a carrier protein for other antigens, particularly monosaccharide antigens, or other antigens lacking a T cell epitope.

The invention belongs to the field of plant engineering technology and crop germplasm resource innovation, and particularly relates to a rice BEIIb gene mutant, application of the rice BEIIb gene mutant in improving rice resistant starch and an improving method. A mutant material with high resistant starch content is created by knocking out the rice BEIIb gene, and compared with a wild type, mutant seeds show an obvious chalky phenotype, composite starch particles are abnormal in a spherical or ellipsoidal shape, the amylose content is remarkably increased, the protein content is remarkably reduced, the resistant starch content is remarkably increased, and the resistant starch content is remarkably reduced. The hardness of cooked rice is remarkably increased, and the elasticity is remarkably reduced. Through transcriptomics analysis, the protein mainly participates in biological pathways such as carbohydrate metabolism, chloroplast photosynthesis, calmodulin combination and secondary metabolite synthesis, and the expression level of endosperm starch synthesis related genes is obviously changed. The method can provide theoretical basis and method reference for breeding of high-quality rice varieties with high resistant starch.

The present disclosure discloses the identification and introduction of a specific heterologous gene encoding an alpha-1,2-fucosyltransferase (denoted smob) into LNT producing strains for the specific production of LNFP-I. The smob gene is derived from the microorganism Sulfuriflexus mobilis (https: / / www.dsmz.de / collection / catalogue / details / culture / DSM-102939), a sulfur-oxidizing bacterium isolated from a saline lake sediment.

The present invention relates to a microorganism which has an enhanced activity of a protein consisting of an amino acid sequence derived from the amino acid sequence represented by SEQ ID NO: 2 by deletion of 20-37 amino acid residues on the N-terminal side, and which has an improved productivity of 3'-sialyllactose compared to the parent strain.

The invention discloses a beta 1, 3-galactosyl transferase Pm beta 3GalT-S207H and an application thereof, and belongs to the technical field of functional enzymes. And the amino acid sequence of the beta 1, 3-galactosyl transferase Pm beta 3GalT-S207H is as shown in SEQ ID NO. 3. The invention further discloses the application of the beta 1, 3-galactosyl transferase Pm beta 3GalT-S207H in the preparation of lactose-N-tetrasaccharide. According to the invention, 1, 3-galactosyl transferase Pm beta 3GalT is subjected to mutation modification to obtain a mutant enzyme, and compared with a wild enzyme, the enzyme activity of the mutant enzyme is improved by 157.21% and is obviously improved, and the thermal stability and alkali resistance are also obviously improved. The beta 1, 3-galactosyl transferase Pm beta 3GalT-S207H disclosed by the invention has important theoretical value and application value on the industrial production of LNT (Lipoxystrobin).

The disclosure provides for recombinantly modified Zika-based vectors that can be used in gene therapy applications.

The invention relates to methods for altering the starch granule composition in starch storage organs. Also described are genetically altered plants characterised by the above phenotype as well as methods of producing such plants.

The present invention relates to a bioconversion for elongating gluco-oligosaccharides with a glucosyl moiety. The present invention also relates to the elongated gluco-oligosaccharides obtained by the process. The elongated gluco-oligosaccharides obtained by the process can be used in edible products, for instance as a dietary fiber or a (low calorie) bulking agent, a prebiotic, in sugar replacement or in calorie reduction. The elongated gluco-oligosaccharides obtained by the process present a decreased digestibility compared to known gluco-oligosaccharides.

The present application relates to the field of bioengineering, and particularly to a mutant of an enzyme, a mutant of a glycosyltransferase and application thereof in preparing rhodiolin. The present application provides a mutant of an enzyme, which has the following amino acid site mutations on the basis of a wild-type glycosyltransferase: the 308th site is mutated from I to Q, the 333rd site is mutated from V to R, the 363rd site is mutated from W to S and the 386th site is mutated from S to G, and the amino acid sequence of the wild-type glycosyltransferase is shown as SEQ ID NO: 1. The present application obtains a glycosyltransferase and a sucrose synthase with improved thermal stability through structural rational design and deep learning modification, and can glycosylate tyramine to generate rhodiolin in the form of regenerating ADP-glucose, and the price of ADP is only about 20% of UDP, which has a cost advantage.