244 results about "Glucuronidase" patented technology

The invention provides a method of modifying soluble xylan which contains glucuronic acid and/or arabinose side chains so that it can be adsorbed onto other substrates. The method includes the steps of enzymatically modifying the xylan by selectively removing glucuronic acid and/or arabinose side chains from the xylan with α-D-glucuronidase and/or α-L-arabinofuranosidase, and allowing the modified xylan to adsorb onto the substrate. The substrate may be a cellulosic substrate like pulp or paper or a non-cellulosic substrate. The enzymes are able to remove a sufficient number of the xylan side chains so as to decrease water solubility of the xylan, induce xylan precipitation and allow adsorption of the xylan onto other substrates. A pulping process which incorporates modification of xylan and adsorption onto cellulosic material is also provided, as are products which contain the modified xylan.

There is disclosed an isolated, modified recombinant β-glucuronidase wherein the modification is having its carbohydrate moeties chemically modified so as to reduce its activity with respect to mannose and mannose 6-phosphate cellular delivery system while retaining enzymatic activity Also disclosed are methods for the treatment of lysosomal storage disease in mammals wherein the mammal is administered a therapeutically effective amount of isolated, modified recombinant β-glucuronidase whereby said storage diseased is relieved in the brain and visceral organs of the mammal. Also disclosed are other lysosomal enzymes within the scope of the invention.

A prodrug utilizes an enzyme whose enzymatic activity is different in between the target site of the drug and the site to express side effects, the prodrug having a substituent cleavable with the enzyme and being activated by cleaving the substituent with the enzyme. As the target site of the drug, for example, a respiratory organ can be mentioned and as the site to express side effects, for example, the heart can be mentioned. As the example of the drug, a bronchodilator can be mentioned and as the example of the enzyme, a glycosidase (for example, β-glucuronidase) can be mentioned. Furthermore, the substituent is, for example, a glycosyl group composed of a monosaccharide or an oligosaccharide. Use of the enzyme enables reducing the side effects of a drug of the type whose target site is different from the site to express side effects.

The invention is relative to a novel method for the detection of coliforms, which is based on the use of a mixture of amino acids to promote the expression of inducible enzymes, in absence of cell growth. This enzyme-inducing solution can be used for the rapid detection of the enzymes β-glucuronidase and/or β-galactosidase in a very small number of E. coli and/or total coliforms.

The invention discloses a method of simultaneously determining contents of 1-OHP ,3-OHB[a]P and 3-OHB[a]A in urine. The method comprises sequentially using beta-glucuronidase for enzymatic hydrolysis of samples, performing solid-phase extraction and purification, concentrating the samples with a vacuum rotary evaporator, and determining by using a liquid chromatography-tandem mass spectrometer, thereby rapidly, accurately and simultaneously detecting contents of 1-hydroxy pyrene (1-OHP), 3-hydroxy benzo [a] pyrene (3-OHB [a] P) and 3-hydroxy benzo [a] anthracene (3-OHB [a] A) in the urine. The method uses deuterated standards as quantitative analysis substances of internal standards and thus can reduce errors in a pretreatment process for the samples; uses a tandem mass spectrometer to relatively improve selectivity and accuracy of the method; and selects a method of preparing standards by matrices, wherein, compared with a method of preparing the standards by pure water, the method of preparing standards by matrices is relatively good in accuracy and can eliminate interference from matrix effects. Through selecting and optimizing chromatographic columns and gradient elution conditions, the method relatively improves a chromatogram separating process and shortens a chromatographic analysis time.

Late embryogenesis abundant (Lea) proteins accumulate in maturing seeds after many of the storage compounds have been synthesized, and they are considered relevant to maturation. We report here the molecular organization and expression of BnLea3-1, a novel Group 3 Lea gene from Brassica napus. BnLea3-1 contains a coding region of 798 bp, sharing 84.4% homology at the amino acid level with Lea76 of B. napus. Two tandem 11-mer repeats are truncated from the coding region of BnLea3-1, compared to the 13 conserved 11-mer repeats of Lea76. Substitutions of consensus residues are found at various positions within the 11-mer repeats. A 1561 bp 5′ flanking promoter fragment of BnLea3-1 fused to E. coli-glucuronidase (GUS) coding region conferred seed-specific GUS expression in stable transgenics of B. napus, tobacco and in transiently-transformed pea. A −137 bp minimal promoter preceding the first transcription start site, identified through progressive deletions from the upstream was sufficient for basal GUS expression in the seeds and in leaves treated with ABA. Deletion studies indicate the presence of enhancing elements located between −137 bp to −742 bp and suppressing elements located between −742 and −1561 bp. BnLea3-1 expression in seeds precedes that of Lea76. Unlike other Group 3 Lea members including HVA1 and Dc3, BnLea3-1 is active in seeds and responsive weakly in vegetative tissues to ABA and methyl jasmonate (MeJA) but not to stress treatments. Possible functions of BnLea3-1 and another member of the Group 3 Lea family BnLea3-2 in embryo development is discussed.

The invention discloses a chromogenic medium used for detecting esherichia coli O157:H7. The medium comprises agar, peptone, beef extract powder, sodium chloride, sorbitol, inositol, neutral red, beta-galactosidase chromogenic substrate, beta-glucuronidase chromogenic substrate, isopropyl-beta-D- thiogalactopyranoside, natrium taurocholicum, potassium tellurite and cefixime. The chromogenic medium used for detecting esherichia coli O157:H7 of the present invention has the advantages of high sensitivity, good specificity, direct bacterial strain discrimination according to colony color, short detection period and strong operationality, is suitable for treating high-reflux samples, is capable of comprehensively, systematically and accurately detecting and preliminarily identifying the esherichia coli O157:H7 in food production and environment, and provides a novel approach for rapidly detecting the microbes.

The present invention provides a novel method for diagnosing, monitoring, prognosing and staging Lymph Node (LN) status in colorectal cancer (CRC) that is more sensitive and accurate than conventional detection technologies such as histopathology. The Guanylyl Cyclase C (GCC) gene is specifically expressed in apical epithelial cells of the GI tract from the duodenum to the rectum and the detection of GCC mRNA in LNs is indicative of the presence of metastases. Quantitative RT-PCR (RT-qPCR) detection of GCC mRNA to identify the presence of colorectal cancer (CRC) cells in LNs has the potential to aid in CRC staging. When used in combination with glucuronidase B (GUSB), accurate quantification of GCC can be achieved with less than a 2-fold variation between intact and highly degraded RNA specimens. The invention also relates to a newly designed GCC/GUSB assay that uses relative quantification having improved prognostic value for time to recurrence and relapse-free survival in Stage I or II colon cancer patients. The GCC/GUSB assay also improves the statistical power of prognosis stratification for relative risk of recurrence and relapse-free survival.

The invention relates to a lubricant which is used for high-speed conveyer apron production lines of PET bottles, glass bottles, ring-pull cans and the like and belongs to the filed of chemical lubricant. The proportion of the water-based chain lubricant is that oleic acid triethanolamine 3-8%, triethanolamine 4-9%, LF-131 2-5%, isothiazolin-ketone 1-3%, polymaleic acid 0.5-2%, glucuronidase 4-9%, EDTA-2Na 4-10%, benzotriazole 0.5-2%, sodium sulfonate xylene 0.5-2% and H2O 65-75%, wherein the water-based chain lubricant has high lubricating property and good solubility and is provided with a plurality of excellent performances as anti-hard water, sterilization, low-foam, non-corrosiveness, crack resistance, anti-coagulation and the like, when the lubricant is used in a lubricating system, the bottles inverting ratio can be effectively reduced, the workshop environment can be improved, and the service life of scraping belts cab be prolonged.

The invention discloses a biological wall breaking technology of pine pollen. The process of the technology comprises the following two steps of: sterilizing pollen and enzymatically breaking the wall of the pollen, wherein the sterilizing time is 12-24 h, the pollen concentration in enzymolysis is 1-50 percent, the enzymatic wall breaking time is 0.5-36 h, and the enzymatic wall breaking temperature is 20-60 DEG C. One or more composite enzymes of the enzymic preparations, such as pepsin, trypsin, cathepsin, papain, subtilisin, lysozyme, Snail enzyme, amylase, arabinosidase, galactosidase, glucuronidase, acetyl xylan esterase, pectase, cellulose and hemicellulase, are used for enzymatic wall breaking. The process is simple and reliable, has less investment and high wall breaking rate, can fully obtain the active ingredients with no loss of activity and keep the original characteristics of the pine pollen.

A thimerosal-free hyaluronidase preparation wherein the preferred hyaluronidase enzyme is devoid of molecular weight fractions below 40,000 MW, between 60-70,000 MW and above 100,000 MW. Also disclosed is a method for accelerating the clearance of hemorrhagic blood from the vitreous humor of the eye, said method comprising the step of contacting at least one hemorrhage-clearing enzyme (e.g., a β-glucuronidase, matrix metalloproteinase, chondroitinase, chondroitin sulfatase or protein kinase) with the vitreous humor in an amount which is effective to cause accelerated clearance of blood therefrom.

The invention relates to a method for improving the instantaneous expression rate of an agrobacterium tumefaciens-mediated sugarcane glucuronidase (GUS). The method comprises the following steps of: selecting materials, preparing conversion bacteria liquid, performing an infection method, performing cocultivation and selective cultivation, and chemically dyeing GUS active tissues. The method that the advantages and disadvantages of a transgenic technology method is measured by taking the degree of the GUS instantaneous expression rate as an index is nationally acknowledged. Therefore, the index which is nationally acknowledged is adopted to evaluate the technology method. By the method, the GUS instantaneous expression rate can be increased by 335.8 to 1,859.3 percent, the bottle neck problem about efficiently obtaining a transgenic sugarcane plant by using a agrobacterium tumefaciens-mediated way can be effectively solved, a key technology method for introducing an exogenous gene into a sugarcane is provided for improving the variety of the sugarcane by using transgenic technology. Meanwhile, the method is also applied to the genetic transformation for other plants based on an agrobacterium tumefaciens-mediated way.

The invention discloses a pseudo-ginseng inducible promoter R1 and application thereof, the nucleotide sequence of R1 is shown as SEQ ID NO: 1, and molecular biology and genetic engineering related technical researches prove that the pseudo-ginseng promoter R1 responds to several plant hormones, biological stress and abiotic stress; a fusion expression cassette constructed by the panax notoginseng promoter R1 and the beta-glucuronidase gene is transferred into tobacco for expression, and the activity of the glucuronidase of the transgenic tobacco is quantitatively detected through a fluorescence method. The result shows that after the transgenic tobacco is treated by fusarium oxysporum, fusarium solani, alternaria alternata, NaCl, AlCl3, gibberellin, methyl jasmonate, salicylic acid, abscisic acid and ethephon, the activity of glucuronidase is obviously enhanced; the pseudo-ginseng promoter R1 is induced by several plant hormones and biological and abiotic stress factors, and can be used for plant disease-resistant gene engineering.

Disclosed are prodrugs of inactivators of O⁶-alkylguanine-DNA alkyltransferase (AGT). The prodrugs are cleavable by the β-glucuronidase enzyme, which is either administered to the patient or produced by necrotic tumor cells. The prodrugs are represented by the formula A-B-C, wherein A is a glucuronosyl residue linked through its 1-oxygen to the phenyl ring of B; B is a benzyloxycarbonyl group, optionally ring-substituted with one or more electron withdrawing groups; and C is an inactivator of AGT, e.g., a substituted or unsubstituted O⁶-benzylguanine or O⁶-benzyl-2′-deoxyguanosine, Also disclosed are additional inactivators of AGT, pharmaceutical compositions comprising an inactivator or prodrug and a pharmaceutically acceptable carrier, and a method of use of the inactivator or prodrug in enhancing the chemotherapeutic treatment of tumor cells in a mammal, e.g., a human, with an antineoplastic alkylating agent that causes cytotoxic lesions at the O⁶-position of guanine.

The invention provides a Dongxiang wild rice endophytic fungus strain capable of transforming glycyrrhizic acid to produce glycyrrhetinic acid monoglucuronide. The strain is identified and named as Aspergillus flavus DX-SEL2 and preserved in China Center for Type Culture Collection, with an accession number of CCTCC NO: M2013686. The strain can induce generation of beta-glucuronidase in a transformation medium with a glycyrrhizic acid as an inductive agent, then hydrolysis is carried out so as to remove a glucuronyl group at the terminal of a glycyrrhizic acid molecule, so glycyrrhetinic acid monoglucuronide is produced (secreted in fermentation broth). Organic solvent extraction, normal-phase silica gel column chromatography, reversed-phase silica gel column chromatography and the like are used for further separation and purification so as to obtain glycyrrhetinic acid monoglucuronide. According to the invention, microbial catalytic conversion is employed, and the advantages of greenness, environmental protection, no pollution, simple transformation steps, a single product, a high transformation rate, etc. are obtained.

The invention discloses a specific expression promoter of a rice seed aleurone layer and application of the rice seed aleurone layer, and provides a deoxyribonucleic acid (DNA) segment which is a DNA molecule shown in 1) or 2) or 3), wherein 1) is the DNA molecule shown in a sequence 1 of a sequence table; 2) is the DNA molecule hybridized with a DNA sequence defined by 1) under a strict condition and having a promoter function; and 3) is the DNA molecule having homology of more than 70 percent with the DNA sequence defined by 1) and having a promoter function. The experiment shows that the promoter P-NF-YB1 is found, rice is introduced, and the expression of glucuronidase (GUS) gene is driven in the rice. The result shows that the rice seed aleurone layer specially expresses that the promoter is a tissue specificity promoter; and the promoter has wide application and market prospect in the field of agriculture.

The invention discloses a method for culturing agrobacterium-mediated transgenic salix matsudana plants and relates to the method for culturing the transgenic salix matsudana plants. The method aims to solve the problems that the existing method cannot induce resistant buds from transgenic transgenices of salix matsudana so that the transgenic salix matsudana plants cannot be obtained. The method comprises the following steps of: firstly, sterilizing and preculturing salix matsudana seeds; secondly, selecting agrobacterium tumefaciens with pBI121 carriers, and culturing to obtain a bacteria solution; thirdly, taking sprouted top bud parts cut from basic parts as explants, and then soaking in the bacteria solution; fourthly, taking out the explants and soaking up through filter papers, and inoculating the explants into a co-culture culture medium; fifthly, switching to a selective medium till the resistant buds grow, switching to a light condition, and screening the resistant buds; sixthly, transferring the resistant buds into an elongation culture medium; seventhly, cutting off stem sections, and switching to a rooting culture medium for culture till roots grow; and finally, carrying out polymerase chain reaction (PCR) and glucuronidase (GUS) staining detection, transplanting to earth, and obtaining the transgenic salix matsudana plants. The method for culturing the agrobacterium-mediated transgenic salix matsudana plants is applied to culturing the transgenic salix matsudana plants.

The invention discloses a method for culturing a transgenic plant with decreased wax. The method for culturing the transgenic plant with the decreasing wax comprises the following steps of: losing the gene coding function of proteins shown in SEQ ID NO: 1 of an initial plant so as to obtain the transgenic plant with the decreased wax. In the method, the gene affects the formation of leaf wax and is the gene for controlling leaf wax to form, and particularly, the gene can positively regulate the formation of leaf wax. The gene has important application value on the understanding of the mechanism of forming the leaf wax of paddy rice, and the gene can be used for breeding paddy rice to improve cuticular properties of paddy rice leaves by the gene engineering method so as to improve the drought control capacity. In addition, the gene and the protein can control the content of cuticular wax which may affect glucuronidase (GUS) staining effect of paddy rice leaves, and the GUS staining effect of paddy rice leaves can be enhanced by lowering the content of cuticular wax, so that the gene can be used to culture transgenic test materials for detecting expression conditions of exogenous gene on leaves.

A recombinant DNA construct is provided and includes a first DNA fragment encoding a β-glucuronidase and a second DNA fragment encoding a membrane anchoring domain. The β-glucuronidase may be a human β-glucuronidase or a mouse β-glucuronidase. In one embodiment, an expression vector for delivering a gene of interest or portion thereof into a host cell includes a DNA sequence for the gene of interest, a first DNA fragment encoding a β-glucuronidase, and a second DNA fragment encoding a membrane anchoring domain. In another embodiment, a method of introducing a gene of interest or portion thereof into a host cell is provided, including introducing into the host cell a recombinant DNA construct.

Mutated Staphylococcus sp. RLH1 β-glucuronidase enzymes with enhanced enzymatic activity and thermostability as compared to wild type enzyme are provided. The enzymes of the invention advantageously allow for accurate analysis of bodily samples for the presence of drugs in 30 minutes or less, as compared to the several hours needed using prior enzyme preparations. Methods of using the mutated enzymes for hydrolysis of glucuronide substrates, including opiates and benzodiazepines, are also provided.

The invention discloses an agrobacterium-mediated spirodela polyrhiza stable transformation system establishment method, which comprises the following steps: A, inducing calli of spirodela polyrhiza; B, connecting a target gene with a T vector; C, connecting the target gene with a plant expression vector; D, transforming the plant expression vector in agrobacterium tumefaciens LBA4404; E, preparing an infective bacteria solution; F, co-culturing the calli and the agrobacterium tumefaciens; G, performing regeneration and selective culturing: determining that the target gene is successfully integrated into a genome of the spirodela polyrhiza if a GUS (glucuronidase) staining result of thalli is blue, and determining that a part which is stained to be blue by thallus tissues is a part expressed by the target gene. The target gene is inserted into the plant expression vector and then transferred into the agrobacterium tumefaciens LBA4404 for callus infection, and the concentration of antibiotics is finally controlled to screen a positive plant to obtain transgenetic spirodela polyrhiza, and an established transformation system can still ensure continuous and stable inheritance after asexual reproduction of the spirodela polyrhiza, and has bio-safety and development and application value.