63 results about "Glucuronidase" patented technology

The invention discloses an application of a tomato B3-337 transcription factor in improvement of tomato quality, a gene for coding a B3-337 transcription factor protein is named as a B3-337 gene, and the nucleotide sequence is shown as SEQ ID NO.1. The invention also discloses an application of the tomato B3-337 transcription factor in improvement of tomato quality. According to the application, a carrier pFM3100GFP-B3-337 is constructed, so that the B3-337 is proved to be positioned in a cell nucleus. A transcriptional activity detection result proves that B3-337 has transcriptional activation activity. RT-qPCR (real-time quantitative polymerase chain reaction) and GUS (glucuronidase) dyeing show that the transcription factor is expressed in pericarp, seed, placenta and center pillar of each fruit ripening stage. According to the invention, a B3-337 gene overexpression plant is obtained by constructing a B3-337 overexpression vector, and compared with a wild type, the tomato fruit overexpressed with the B3-337 gene has the advantages that the content of soluble solids brix is increased, the fruit becomes larger, and the weight becomes heavier. It is shown that the B3-337 transcription factor plays an important regulation role in positive regulation of tomato fruit quality. The invention lays a good gene genetic resource foundation for the cultivation of new varieties of high-quality tomatoes.

The invention belongs to the technical field of biology, and discloses a gene of beta-glucuronidase, an amino acid sequence, an expression vector and a host cell of the beta-glucuronidase and application of the beta-glucuronidase in preparation of glycyrrhetinic acid monoglucuronide. The gene of the beta-glucuronidase is connected with an expression vector and is transformed into a saccharomyces cerevisiae cell, so that the saccharomyces cerevisiae engineering strain with high yield of the beta-glucuronidase is successfully constructed. The glycyrrhizic acid can generate a single product glycyrrhetinic acid monoglucuronide under the catalytic action of the enzyme, and no by-product is generated, so that the aim of efficiently, specifically and safely preparing glycyrrhetinic acid monoglucuronide is fulfilled, and the application and development prospect is wide.

The invention provides a complex microbial inoculant for producing beta-glucuronidase and application of the complex microbial inoculant in calculus bovis production, and relates to the technical field of microorganisms. The culture method of the complex microbial inoculant provided by the invention comprises the following steps: culturing lactobacillus ATCC 53103 in an initial culture medium to a logarithmic phase, sterilizing to obtain a complex culture medium, mixing the complex culture medium with nutritional ingredients, inoculating escherichia coli ATCC 25922 and riemerella CCUG 14215, and co-culturing to obtain the complex microbial inoculant. The complex microbial inoculant provided by the invention can generate a large amount of high-activity beta-glucuronidase, so that the efficiency of in-vitro production of calculus bovis can be effectively improved.

The invention discloses a tissue-specific promoter screening vector and application thereof, and relates to the technical field of plant genetic engineering. The screening vector comprises an expression vector and a modified GUS (glucuronidase) reporter gene connected to the expression vector, the modified GUS reporter gene is Cat1muGUS, and the sequence of the Cat1muGUS is shown as SEQ ID NO. 3. According to the present invention, the Cat1muGUS variant is developed based on the Cat1GUS, the Cat1muGUS sensitivity is high, the problem of low dyeing screening efficiency caused by the fact that the Cat1GUS is not sheared in the dicotyledonous plant and the monocotyledonous special tissue can be overcome, in addition, the tissue-specific promoter screening vector is constructed based on the Cat1muGUS variant, and the screening efficiency is high; the method can be well applied to screening of all plant and all tissue specific promoters, and the screening efficiency is improved.

The invention provides a method for producing beta-glucuronidase from escherichia coli and application of the beta-glucuronidase, and relates to the technical field of microorganisms. The method provided by the invention comprises the following steps: inoculating Escherichia coli ATCC 25922 on a composite culture medium for pre-culture, and culturing in an electric field environment of 1-10N / C for 12-24h to obtain a mixture containing beta-glucuronidase; wherein the composite culture medium comprises 10-100 [mu] mol / L of ferric citrate, and the concentration of the ferric citrate is increased in stages in the pre-culture process. According to the method, the escherichia coli ATCC 25922 is cultured in the iron-containing culture medium and the low-intensity electric field environment, so that the synthesis of the beta-glucuronidase by the escherichia coli ATCC 25922 can be effectively improved, and the enzyme activity and the enzyme yield of the beta-glucuronidase are improved.

The present invention discloses a method for synchronously extracting and detecting 59 endocrine disruptors in amniotic fluid, belonging to the technical field of substance detection. The method includes the following steps: S1, adding β-glucuronidase to the amniotic fluid sample and incubating; S2, adding a stable isotope-labeled internal standard to the amniotic fluid sample to be detected in step S1; S3, adding the amniotic fluid sample to be detected in step S2 to an activated HLB extraction column to remove impurities and extract the target analyte; S4, eluting the extraction column to obtain an eluate, blowing it to near dryness, and adding a reconstitution solvent to obtain a sample to be detected; S5, using a liquid chromatography-mass spectrometry instrument to detect the sample to be detected obtained in step S4 to complete the detection and analysis of endocrine disruptors. The detection process is carried out simultaneously in positive ion mode and negative ion mode. This method realizes the synchronous extraction, detection and analysis of at least 59 typical environmental endocrine disruptors in amniotic fluid, which is simple and convenient, and solves the problems of large sample consumption and long time consumption.

The invention discloses a lutein composition for relieving asthenopia and a preparation method of the lutein composition, and belongs to the field of functional foods. Phycocyanin and kelp polyphenol are mixed and react to obtain modified phycocyanin; fucoidin and beta-glucuronidase are mixed and react to obtain modified fucose; mixing the fish collagen peptide and zinc chloride to react to obtain a collagen peptide-zinc chelate; the xanthophyll siliceous cage compound and the modified phycocyanin are mixed and react to obtain a first compound; mixing and reacting the first compound and modified fucose to obtain a second compound; mixing and reacting the second compound, collagen peptide-zinc chelate and krill oil to obtain a lutein compound; mixing and reacting the second compound, collagen peptide-zinc chelate and krill oil, adding trehalose, and mixing and reacting to obtain a xanthophyll compound; and mixing the xanthophyll compound and maltodextrin to obtain the xanthophyll composition.

The invention discloses a beta-glucuronidase mutant and application thereof, and belongs to the technical field of genetic engineering and protein engineering modification. The beta-glucuronidase mutant disclosed by the invention has the following mutation on the basis of an amino acid sequence as shown in SEQ ID NO.3: T512S or T512A. The catalytic efficiency of the beta-glucuronidase mutant T512S is improved by 13.92 times compared with that of AtGUS (-3t), and the beta-glucuronidase mutant T512S shows remarkable industrial application potential.

Provided is a glucuronidase activator containing, as an active ingredient, Levilactobacillus brevis (Levilactobacillusbrevis) strain I-3141 (accession number: CECT 7480).

The present disclosure provides an engineered enzyme comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of a human beta-glucuronidase (hBG). Also provides a fusion protein comprising a polypeptide and the engineered enzyme, and a method of enhancing an anticancer activity of an anticancer drug in a subject.

Provided herein are plants, plant parts, a population of plants or plant parts, and plant products (e.g., seed composition, protein composition) comprising reduced activity of a protein-related polypeptide [e.g., stomatal cytokinesis defective 2 (SCD2), SCD2A, SCD2B, response to dehydration 22 (RD22), glucuronidase 3 (GUS3), GUS3A, glycosyl hydrolase family 10 protein B (GH10B), protein phosphatase 2A beta subunit (PP2AB), PPA2BA, PP2ABAB, alpha / beta-hydrolases superfamily protein (ABH), ABHA, ABHB, calmodulin-binding transcription activator protein 2 (CAMTA2), CAMTA2A, CAMTA2B, cinnamyl-alcohol dehydrogenase (CAD1), beta-ketoacyl reductase 1 (KCR1), KCR1A, or KCR1B], and compositions and methods of producing such plants and plant parts. The plants, plant parts, population of plants or plant parts, or plant products can have a genetic mutation that reduces the protein-related polypeptide activity, which can be one located at least partially in a protein-related gene (e.g., CAD1) or its homolog or in its regulatory region, and can have increased protein content and / or white flake protein content.

The invention discloses an oxgall-tolerant in-vitro calculus bovis culture transformation strain and application thereof, and belongs to the technical field of calculus bovis culture. The strain is obtained by separating and purifying fresh ox bile, is identified to belong to escherichia coli, has excellent ox bile tolerance, has the characteristics of beta-glucuronidase, high bezoar conversion rate and high safety, and can be used in the production of in-vitro artificial cultivation of bezoar. The strain is a brand new strain for in-vitro calculus bovis transformation, not only enriches a strain resource library of calculus bovis transformation bacteria, but also provides a new thought for deep research of subsequent in-vitro calculus bovis culture.

The present invention discloses a selenium-rich probiotic that affects irinotecan-induced diarrhea and intestinal flora in mice, establishes an irinotecan-induced diarrhea mouse model, and administers selenium-rich probiotics by long-term oral gavage to evaluate its effects on irinotecan-induced diarrhea and intestinal flora in mice. The mice in the experimental group were given selenium-rich probiotics by oral gavage every day, and the weight, diarrhea grade, immune index, intestinal flora and ileum tissue structure of the mice were measured. The results show that selenium-rich probiotics can inhibit weight loss and reduced immune function caused by irinotecan; reduce the incidence of diarrhea and the average diarrhea grade in mice, and alleviate irinotecan-induced diarrhea in mice; reduce the content of Clostridium, Escherichia coli and Bacteroidetes associated with β-D-glucuronidase, and increase the content of bifidobacteria in the intestine; alleviate the damage of intestinal mucosa, cells and glands of mice caused by irinotecan.

A glucuronidase activator including an active ingredient that is a Levilactobacillus brevis strain I-3141 deposited under accession number CECT 7480.

This invention provides an in vitro cultured bezoar, its preparation method, and its application, belonging to the field of traditional Chinese medicine preparation technology. The preparation method of this invention involves obtaining a bile-resistant, high-β-glucuronidase-active *Escherichia coli* strain through bile gradient acclimation, which is then combined with *Enterococcus faecalis* to form a functional bacterial community. This community is then cultured in vitro through a three-stage process of proliferation, transformation, and crystallization, followed by gradient calcium ion-induced crystallization and vacuum freeze-drying to obtain in vitro cultured bezoar. This invention aims to solve the problems of low conversion efficiency, uneven crystallization, insufficient content of effective components, and unstable product quality in existing technologies. The obtained in vitro cultured bezoar product has significantly higher bilirubin and conjugated bile acid contents than the standards of the *Chinese Pharmacopoeia*, and the process is stable, controllable, and suitable for industrialization.

The application discloses a kind of in-vivo cultivation of bovine calculus bezoar gel and preparation method thereof, it is related to in-vivo cultivation of bovine calculus bezoar technical field.A kind of in-vivo cultivation of bovine calculus bezoar gel, including following mass component: with mass percentage, 0.1%~5% drug depot, 8%~20% high molecular gel skeleton, 0.1%~2% crosslinking agent, 0.1%~2% crosslinking regulator, 50%~65% solvent, 10%~40% crosslinking agent solution;Drug depot exists in the form of β-glucuronidase sustained-release microsphere.The sponge-like porous gel prepared by the application can be injected into the gallbladder of a cow to form a sponge-like bovine calculus bezoar bed through crosslinking reaction, the bovine calculus bezoar bed can slowly release β-glucuronidase, promote the decomposition of conjugated bilirubin into free bilirubin, and finally the free bilirubin forms calcium bilirubinate, which, after combining with mucin, forms bovine calculus bezoar particles, and the particles are enriched by the bovine calculus bezoar bed to form bovine calculus bezoar with high yield and good quality.

A glucuronidase activator including an active ingredient that is a Levilactobacillus brevis strain I-3141 deposited under accession number CECT 7480.

The invention discloses a method for detecting various pollutants in follicular fluid. The method comprises the steps of enzymolysis, extraction, nitrogen blowing, redissolution and detection on a machine. Mixing the follicular fluid sample with the mixed internal standard solution, performing enzymolysis overnight, and hydrolyzing the bound metabolite by using beta-glucuronidase; extracting by using an ethyl acetate / methyl tert-butyl ether mixed solution containing 0.1% formic acid, and repeating for three times to extract 13 types of 182 pollutants; and carrying out nitrogen blowing drying and acetonitrile / water redissolution, and analyzing through a series quadrupole rod liquid chromatograph-mass spectrometer. According to the method, the detection limit is as low as nanogram / milliliter, quantification is completed within a short time through single sample injection, more than 100 samples can be detected in a single day, high-throughput and high-sensitivity detection is achieved, energy consumption and cost are reduced through program optimization, high efficiency and environmental friendliness are achieved, and a powerful means is provided for studying the follicular fluid pollutant exposure level and correlating with reproductive health.

The invention relates to the technical field of beta-glucuronidase production, and discloses recombinant escherichia coli for expressing beta-glucuronidase and application of the recombinant escherichia coli. According to the method, beta-GD derived from clostridium perfringens is optimized by preferred codons and then is introduced into escherichia coli to obtain recombinant escherichia coli for fermentation, and the beta-GD with high activity is separated from thalli of the recombinant escherichia coli. Specifically, the yield of beta-GD expressed by the gene reaches 116 mg / L, and the activity of the prepared beta-GD reaches 664 U / mg. Therefore, the problem of low activity of beta-GD obtained by existing escherichia coli expression is solved. The amino acid sequence of the beta-GD derived from clostridium perfringens is as shown in SEQ ID No. 1, and the nucleotide sequence of the beta-GD derived from clostridium perfringens is as shown in SEQ ID No. 2.

The present application relates to the technical field of drug residue detection, and in particular to a general pre-treatment method based on immunoaffinity rapid detection. A general pre-treatment method based on immunoaffinity rapid detection, comprising the following steps: using reagent 1, reagent 2, reagent 3, reagent 4, reagent 5 to process the sample to be tested in sequence, wherein the reagent 1 is ammonium citrate buffer; the reagent 2 is β-glucuronidase; the reagent 3 is arylsulfatase; the reagent 4 is a mixture of acetonitrile, methanol, ethanol and an organic acid; the organic acid is citric acid or formic acid; the reagent 5 is N-propylethylenediamine. The pre-treatment method of the present application is applicable to the pre-treatment of the residue detection of multiple major drugs involved in veterinary drug residues in current animal food, and solves the problem that food safety detection usually requires the determination of multiple drugs on the same sample, and multiple pre-treatment operations are required, and the time, equipment and reagent consumption are large.

Provided are a β-glucuronidase, a recombinant expression vector thereof, an engineering bacterium, a fermenting agent, and a mass production method for glycyrrhetinic acid. The provided β-glucuronidase is selected from β-glucuronidase AcGUS, β-glucuronidase AcGUS1, β-glucuronidase AcGUS2, β-glucuronidase AcGUS3, and a β-glucuronidase AcGUS3 mutant. The β-glucuronidase AcGUS is an enzyme obtained in such a way that in the amino acid sequence with the GenBank accession number of AEK69352.1, the amino acid on the 1st site is deleted, the alanine on the 7th site is mutated into glycine, the arginine on the 66th site is mutated into lysine, the alanine on the 256 site is mutated into valine, the threonine on the 270 site is mutated into alanine, the valine on the 332nd site is mutated into phenylalanine, the aspartic acid on the 363rd site is mutated into glutamic acid, and the methionine on the 506th site is mutated into valine. The provided β-glucuronidase can be used for industrial scale production of 18α-GA and 18β-GA.

The invention relates to the technical field of analysis and detection, in particular to a detection method for quantitative urine steroid hormone based on efficient enzymolysis. The invention establishes a detection method for accurately quantifying 35 steroid hormones in urine by combining a high performance beta glucuronide enzymatic dissociation method with high performance liquid chromatography-tandem mass spectrometry. According to the method, the pretreatment time required by clinical determination of urine steroid hormones can be remarkably shortened, single-needle quantitative detection of 35 steroid hormones is realized, and the method has a wide application prospect.

The invention relates to Leclercia adcarboxsaca and an application thereof in degradation of corn straws, the Leclercia adcarboxsaca is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 32394. The strain can secrete AA2 family DyP-type peroxidase, GH1 / GH3 beta-glucosidase, PL8 oligogalacturonic acid lyase and GH88 unsaturated glucuronic acid hydrolase, and has the capability of synergistically degrading lignocellulose. The culture method comprises the steps of activation, seed solution preparation and fermentation. The obtained fermentation liquor is applied to the corn straw in proportion, so that the surface structure of the straw can be effectively destroyed, and the degradation rate is increased; meanwhile, soil nutrients are remarkably activated, the content of rapidly available phosphorus, rapidly available potassium, soluble organic carbon, alkali-hydrolyzable nitrogen and the like in the soil is greatly increased, and the synergistic interaction of efficient straw degradation and soil fertility improvement is achieved.

This invention discloses a lutein composition for relieving eye fatigue and its preparation method, belonging to the field of functional foods. The process involves: reacting phycocyanin and kelp polyphenols to obtain modified phycocyanin; reacting fucoidan and β-glucuronidase to obtain modified fucose; reacting fish collagen peptides and zinc chloride to obtain collagen peptide-zinc chelate; reacting lutein silica cage complex and modified phycocyanin to obtain a first complex; reacting the first complex and modified fucose to obtain a second complex; reacting the second complex, collagen peptide-zinc chelate, and krill oil to obtain a lutein complex; reacting the second complex, collagen peptide-zinc chelate, and krill oil, then adding trehalose to obtain the lutein complex; and finally mixing the lutein complex with maltodextrin to obtain the lutein composition.

The invention discloses a chlorinated spiro-gentianol derivative as well as a preparation method and application thereof, and belongs to the technical field of active ingredient analysis of marine fungi. The chlorinated spiro-gentianol derivative is a derivative with a chlorogentisyl alcohol and gbosine heterozygous skeleton, the chlorinated spiro-gentianol derivative with a novel structure is separated from a fermentation culture of marine fungi Epicoccum sorghine.GST-5 by utilizing the polarity difference of the chlorogentisyl alcohol and the gbosine derivative, and through an in-vitro anti-beta-glucuronidase test, the chlorinated spiro-gentianol derivative with the novel structure can be used for preparing the anti-beta-glucuronidase derivative with the novel structure and the application of the anti-beta-glucuronidase derivative in the marine fungi Epicoccum sorghine.GST-5, so that the anti-beta-glucuronidase derivative with the novel structure is obtained. It is shown that the chloro-spiro-gentianol derivative provided by the invention has good beta-glucuronidase inhibitory activity, is low in cytotoxicity, and has development prospects in the aspect of preparation of anti-beta-glucuronidase drugs or functional foods.

The application belongs to the technical field of bioengineering, and specifically discloses a glucuronidase and a mutant thereof and application of the glucuronidase and the mutant in production of bilirubin. The glucuronidase is derived from Escherichia coli, and an amino acid sequence of the glucuronidase is shown as SEQ ID NO. 1. An amino acid sequence of the mutant of the glucuronidase is shown as SEQ ID NO. 2, and the mutant is obtained by truncating amino acids 360-376 in the amino acid sequence of the glucuronidase. A coding gene of the glucuronidase or the mutant thereof is connected with an expression plasmid to construct an expression vector, the expression vector is introduced into a host strain to obtain a recombinant strain, and the recombinant strain is cultured to obtain whole-cell catalytic bacteria slurry. The recombinant strain or the whole-cell catalytic bacteria slurry can be used in preparation of bilirubin, has a high substrate conversion rate and less by-products, the preparation method is simple and convenient, production conditions are mild, environmental pollution is small, and the application has a good technical application prospect.

The invention discloses a method for predicting fruit color based on structural variation of a strawberry FaMYB10-2 promoter, variation analysis is carried out on FaMYB10-2 promoter regions of 200 strawberry varieties to find that significant structural variation (SV) exists at the position 986bp away from the upstream of an initiation codon (ATG) on the promoter, the SV is in high linkage imbalance with adjacent SNP, and the SV is in high linkage imbalance with adjacent SNP. Promoter variation causes significant difference of FaMYB10-2 gene expression levels, and further influences anthocyanin synthesis and fruit color shade. Ref / Ref genotypes are mainly distributed in strawberry varieties bred in European and North America, and the fruit color is relatively deep; alt / Alt genotypes are mostly found in strawberry varieties bred in China and Japan, and the fruit color is light. Proved by activity analysis of promoter-driven GUS (glucuronidase), the Ref genotype promoter has strong activity, while the Alt genotype promoter has weak activity. The molecular marker is designed by using the structural variation and the related SNP, so that the rapid typing and auxiliary breeding of the fruit color character can be realized. The invention provides a new understanding for a molecular regulation mechanism of strawberry fruit color, and has wide breeding application value.

The present invention discloses a β-glucuronidase and its application in bilirubin production, belonging to the field of biomedicine technology. The present invention obtains a product containing β-glucuronidase by constructing a recombinant plasmid, transforming, expressing and isolating and purifying, which can be used to prepare bilirubin by pig bile reaction. The β-glucuronidase provided by the present invention has high enzyme activity and thermal stability, a wide pH range and temperature range, and strong adaptability. In addition, it contains Fe 2+ and Mg 2+ The metal ion solution has a promoting effect on β-glucuronidase, which is beneficial for improving enzyme activity. The β-glucuronidase produces bilirubin with high yield and purity and can be widely used in the production process of bilirubin.

The utility model relates to a glucuronidase separating device which comprises a centrifugal machine body and a base installed at the bottom of the centrifugal machine body in a sliding mode. The two sides of the base are each provided with an adsorption piece used for firmly placing the base. A buffering piece used for reducing vibration transmission of the centrifugal machine body is installed in the base. The buffer part comprises four push rods hinged to the bottom of the centrifugal machine body, the centrifugal machine body is started to work, generated vibration is transmitted to the four push rods, the four push rods push the moving rod to move, the piston block at one end of the moving rod reciprocates in the sealing frame, the sealing frame is filled with damping oil, and in the moving process of the piston block, the piston block is driven by the damping oil. Damping oil in the sealing frame circulates through the circulation holes in the surface of the piston block, so that large damping force is generated, resistance borne by the piston block in the vibration process can enable the vibration amplitude of the piston block to be gradually reduced, and therefore energy absorption buffering is conducted on vibration generated during work of the centrifugal machine body.