83 results about "Hydrolase activity" patented technology

The present invention relates to monoclonal antibodies that bind to the extracellular domain of prostate-specific membrane antigen (PSMA), hybridoma cell lines producing the antibodies, and methods of using such antibodies for diagnosis and treatment of cancer. In particular, thirty-five monoclonal antibodies reactive with PSMA expressed on the cell surface are exemplified. Additionally, the present invention relates to a novel protein variant (PSM′) of the PSMA detected by a number of the antibodies of the invention. The hydrolase activity of PSMA and PSM′ allows the use of an immunoenzymatic assay for their detection.

The preparation method to extract tea saponin from oil meal, which comprises smashing, removing enzyme, leaching with hot mixing to boost infiltration capacity, centrifugal solid-liquid separating to add flocculant, deposition separating, disc centrifugal separating for cleaning solution, prefiltration separating for flocculation deposition, nano-membrane concentrating, and spray drying. Wherein, using chemical treatment to inhibit and eliminate hydrolase activity before leaching. This invention is simple and low cost with yield more than 70% and 80% purity.

The present invention relates to monoclonal antibodies that bind to the extracellular domain of prostate-specific membrane antigen (PSMA), hybridoma cell lines producing the antibodies, and methods of using such antibodies for diagnosis and treatment of cancer. In particular, thirty-five monoclonal antibodies reactive with PSMA expressed on the cell surface are exemplified. Additionally, the present invention relates to a novel protein variant (PSM′) of PSMA detected by a number of the antibodies of the invention. The hydrolase activity of PSMA and PSM′ allows the use of an immunoenzymatic assay for their detection.

The invention relates to polypeptides having glucanase, e.g., endoglucanase, mannanase, xylanase activity or a combination of these activities, and polynucleotides encoding them. In one aspect, the glucanase activity is an endoglucanase activity (e.g., endo-1,4-beta-D-glucan 4-glucano hydrolase activity) and comprises hydrolysis of 1,4-beta-D-glycosidic linkages in cellulose, cellulose derivatives (e.g., carboxy methyl cellulose and hydroxy ethyl cellulose) lichenin, beta-1,4 bonds in mixed beta-1,3 glucans, such as cereal beta-D-glucans or xyloglucans and other plant material containing cellulosic parts. In addition, methods of designing new enzymes and methods of use thereof are also provided. In alternative aspects, the new glucanases e.g., endoglucanases, mannanases, xylanases have increased activity and stability, including thermotolerance or thermostability, at increased or decreased pHs and temperatures.

The soporific activity of cis-9,10-octadecenoamide and other soporific fatty acid primary amides is neutralized by hydrolysis in the presence of fatty-acid amide hydrolase (FAAH). Hydrolysis of cis-9,10-octadecenoamide by FAAH leads to the formation of oleic acid, a compound without soporific activity. FAAH has be isolated and the gene encoding FAAH has been cloned, sequenced, and used to express recombinant FAAH. Inhibitors of FAAH are disclosed to block the hydrolase activity.

The present invention includes compositions and methods for treating disease and disorders associated with pathological calcification or pathological ossification by modulating the level or activity of NPP1 or a mutant thereof, or a mutant NPP4 modified to exhibit ATP hydrolase activity similar to the hydrolase activity of NPP1.

The invention relates to polypeptides having glucanase, e.g., endoglucanase, mannanase, xylanase activity or a combination of these activities, and polynucleotides encoding them. In one aspect, the glucanase activity is an endoglucanase activity (e.g., endo-1,4-beta-D-glucan 4-glucano hydrolase activity) and comprises hydrolysis of 1,4-beta-D-glycosidic linkages in cellulose, cellulose derivatives (e.g., carboxy methyl cellulose and hydroxy ethyl cellulose) lichenin, beta-1,4 bonds in mixed beta-1,3 glucans, such as cereal beta-D-glucans or xyloglucans and other plant material containing cellulosic parts. In addition, methods of designing new enzymes and methods of use thereof are also provided. In alternative aspects, the new glucanases e.g., endoglucanases, mannanases, xylanases have increased activity and stability, including thermotolerance or thermostability, at increased or decreased pHs and temperatures.

The present invention concerns a new β-galactosidase with transgalactosylating activity isolated from Bifidobacterium bifidum and a truncated enzyme where the C-terminal end of the β-galactosidase protein has been deleted, resulting in an enzyme with a higher transgalactosylating activity than hydrolase activity. When lactose is used as a substrate, galacto-oligosaccharides are products of the transgalactosylase activity. Galacto-oligosaccharides enhance growth of health-promoting Bifidobacterium that may be used in a number of applications in the dairy industry.

The release by trichomonads of a hydrolase that hydrolyzes a narrowly defined class of substrates at a low pH without interference from hydrolases that are unrelated to trichomoniasis is the basis for a selective diagnostic assay for trichomoniasis that measures hydrolysis of any of these substrates by vaginal fluid at a low pH. Selective assays for trichomoniasis are also obtained by removing particulate matter from a sample of vaginal fluid to extract a fraction devoid of particles greater than a selected size, and where desired, combining the extracted fraction with any of certain specified hydrolase inhibitors, then testing the fraction for enzymatic hydrolase activity. These qualities of trichomoniasis are the basis for a series of diagnostic tests and test devices that produce results that are detectable by visual and other means with a high degree of accuracy.

Recombinant bacteria capable of producing glycerol and glycerol-derived products from sucrose are described. The recombinant bacteria comprise in their genome or on at least one recombinant construct: a nucleotide sequence encoding a polypeptide having sucrose transporter activity; a nucleotide sequence encoding a polypeptide having fructokinase activity; and a nucleotide sequence encoding a polypeptide having sucrose hydrolase activity. These nucleotide sequences are each operably linked to the same or a different promoter. These recombinant bacteria are capable of metabolizing sucrose to produce glycerol and / or glycerol-derived products such as 1,3-propanediol and 3-hydroxypropionic acid.

The present invention relates to monoclonal antibodies that bind to the extracellular domain of prostate-specific membrane antigen (PSMA), hybridoma cell lines producing the antibodies, and methods of using such antibodies for diagnosis and treatment of cancer. In particular, thirty-five monoclonal antibodies reactive with PSMA expressed on the cell surface are exemplified. Additionally, the present invention relates to a novel protein variant (PSM′) of PSMA detected by a number of the antibodies of the invention. The hydrolase activity of PSMA and PSM′ allows the use of an immunoenzymatic assay for their detection.

The invention relates to polypeptides having glucanase, e.g., endoglucanase, mannanase, xylanase activity or a combination of these activities, and polynucleotides encoding them. In one aspect, the glucanase activity is an endoglucanase activity (e.g., endo-1,4-beta-D-glucan 4-glucano hydrolase activity) and comprises hydrolysis of 1,4-beta-D-glycosidic linkages in cellulose, cellulose derivatives (e.g., carboxy methyl cellulose and hydroxy ethyl cellulose) lichenin, beta-1,4 bonds in mixed beta-1,3 glucans, such as cereal beta-D-glucans or xyloglucans and other plant material containing cellulosic parts. In addition, methods of designing new enzymes and methods of use thereof are also provided. In alternative aspects, the new glucanases e.g., endoglucanases, mannanases, xylanases have increased activity and stability at increased pH and temperature.

The present invention includes compositions and methods for treating disease and disorders associated with pathological calcification or pathological ossification by modulating the level or activity of NPP1 or a mutant thereof, or a mutant NPP4 modified to exhibit ATP hydrolase activity similar to the hydrolase activity of NPP1.

The invention provides pediococcus acidilactici which is classified and named (Pediococcus acidilactici) CY-10 and conserved in the general microbiology center of China committee for culture collection of microorganisms, the address is number three in courtyard number one on Beichen west road in the Chaoyang District of Beijing, the postcode is 100101, the conservation series number is CGMCC NO.12450, and the conservation date is May,13th, 2016. The invention further provides application of pediococcus acidilactici in kitchen waste treatment. Pediococcus acidilactici has high protease, amylase and lipid hydrolase activity, degrades glucose to produce acid, has thermophilic bacteria with high salt and acid resistance and can effectively improve the effect of treating other kinds of kitchen waste, and the weight decrement and synergism reach 5% or more.

The fluorescent detecting method for nitrile hydrolase activity includes hydrolyzing fluorescent probe with nitrile hydrolase and measuring the fluorescence strength to obtain nitrile hydrolase activity data. The fluorescent probe is made of salicyl nitrile compound in the structure as shown. The present invention has the advantages of simple preparation of fluorescent probe, stable property and high purity of the fluorescent probe, and high detection sensitivity, precision and speed.

The invention provides a process of producing bio-available folate, i.e. folic acid having an increased proportion of monoglutamyl folate and a decreased proportion of polyglutamyl folate, by culturing micro-organisms containing an active heterologous or homologous polyglutamyl hydrolase activity or containing increased activities of folate biosynthesis enzymes. The genes encoding the polyglutamyl hydrolase and the folate biosynthesis enzymes may be of various origin, e.g. from rodents or other mammals including man. Also provided is a foodstuff, especially a dairy product containing such monoglutamyl folate-producing micro-organisms.

The invention provides mature peptide of a myotis brandti leukotriene A4 hydrolase inhibitor Motistin and an application of the mature peptide, and belongs to the technical field of biological medicine. The amino acid sequence of the mature peptide is shown as SEQ ID NO: 2. The invention discovers for the first time that the mature peptide of Motistin can inhibit the hydrolase activity of LTA4H without affecting the aminopeptidase activity of LTA4H. The mature peptide of Motistin can target LT44H, can inhibit the recruitment of immune cells and the generation of inflammatory mediators by inhibiting the hydrolase activity of LTA4H, and also has the function of inhibiting the generation of inflammatory cytokines IL6, TNF <alpha> and IL1<beta> caused by virus and bacterial lipopolysaccharide.The mature peptide has the potential of becoming an effective drug for inhibiting inflammation.

The invention discloses a fluorescence detection method for hydrolase activity. The method comprises the following steps: hatching detection liquid containing a substrate and a Cu<2+>-bathocuproine disulfonic acid complex with a to-be-detected hydrolase solution, taking the Cu<2+>-bathocuproine disulfonic acid complex as a fluorescent probe, and reducing the fluorescent probe by a reducing product obtained by hydrolyzing the substrate by virtue of hydrolase, so as to generate the Cu<2+>-bathocuproine disulfonic acid complex and weaken a fluorescent signal of the detection liquid. The higher is the activity of the hydrolase in a sample, the lower fluorescence emission intensity of the detection liquid is. The fluorescence detection method disclosed by the invention has high detection sensitivity on hydrolase activity, good selectivity, strong interference resistance, good reproducibility, simple operation and short detection time and can be used for quick and high-sensitivity detection of the hydrolase activity in a clinical sample, and the detection cost is greatly reduced.

An object of the present invention is to provide a PHA-producing microorganism which can assimilate sucrose, and a method for producing a PHA by culturing this microorganism, using sucrose as a carbon source. A PHA-producing microorganism, comprising a PHA synthase gene, and heterogeneous-organism-derived genes in the following items (1) and (2): (1) a sucrose hydrolase gene encoding an amino acid sequence described in SEQ ID NO: 1, or a gene encoding a polypeptide which has a sequence homology of 90% or more to the amino acid sequence and which has sucrose hydrolase activity; and (2) a sucrose permease gene encoding an amino acid sequence described in SEQ ID NO: 2, or a gene encoding a polypeptide which has a sequence homology of 90% or more to the amino acid sequence and which has sucrose permease activity. The invention is also a method for producing a PHA, including the step of culturing this microorganism in a medium including sucrose as a carbon source.

The invention discloses a method for preparing ginsenosides of Gyp XVII, compound O, Mb and F2. The method comprises the following steps that panoxadiol saponins as substrates and a protein undergo a catalytic reaction to produce reactants containing ginsenosides of Gyp XVII, compound O, Mb and F2, wherein the protein is a protein (a) or (b), the protein (a) has an amino acid sequence shown in the formula of SEQ ID No.2 or shown in the formula of SEQ ID No.4, and the protein (b) is derived from the amino acid sequence of the protein shown in the formula of SEQ ID No.2 or shown in the formula of SEQ ID No.4 by replacement and / or deletion and / or addition of one or more amino acid residues and has ginsenoside hydrolase activity. An experiment proves that the method for preparing ginsenosides of Gyp XVII, compound O, Mb and F2 has good practicality.

Glycoside hydrolases having at least two different hydrolytic activities are provided. In one embodiment, an isolated recombinant hydrolase having at least two activities selected from a group including asparagine derivatives, glutamine derivatives, and histidine derivatives is provided. Further, a method of generating free sugars from a mixture comprising asparagine derivatives, glutamine derivatives, and histidine derivatives is provided.

The invention belongs to the technical field of biochemical analysis and detection, in particular to a detection method and a detection kit for acid hydrolase activity in lysosomes. The present invention firstly designs a detection system comprising 5 kinds of mucopolysaccharide acid hydrolase in lysosomes, these five kinds of enzymes are respectively: α-L-iduronidase, iduronate sulfatase, β-N-acetyl half Lactosamine‑6‑sulfatase, β‑galactosidase and arylsulfatase B enzymes. The present invention utilizes the characteristic of fluorescence of 4-methylumbelliferone, uses the new substrate produced by combining the 4-methylumbelliferone group obtained by chemical synthesis with the enzyme substrate, and adopts the acid buffer solution containing the new substrate and mucopolysaccharide The hydrolase reacts, and the free 4-methylumbelliferone fluorescent substance is released after the hydrolase hydrolyzes the substrate. The higher the activity of the hydrolase in the sample, the more the amount of 4-methylumbelliferone released by hydrolyzing the substrate, and the higher the fluorescence intensity generated by the fluorescence detection, so as to realize the quantitative detection of the hydrolase activity. The invention has high detection sensitivity, good selectivity, strong anti-interference ability, good reproducibility and easy operation.

The invention discloses a biocontrol bacterial strain CH01 for efficiently preventing and treating citrus canker and an application of the biocontrol bacterial strain CH01. The biocontrol bacterial strain CH01 is bacillus cereus, and is assigned with the preservation number CGMCC NO.11677. It is shown from a flat inhibition test result the bacillus cereus CH01 can obviously inhibit growth of xanthomonas axonopodis pv. citri, and has an obvious inhibition effect on multiple pathogenic fungi. Further, the bacillus cereus CH01 shows great hydrolase activity in vitro. It is shown from a greenhousetest result that the bacillus cereus CH01 can effectively prevent development of citrus canker, and it is shown from a field trial result that the bacillus cereus CH01 has 65.46% of prevention effecton citrus canker. Therefore, the biocontrol bacterial strain CH01 can be used as a biocontrol preparation and efficiently prevent citrus canker.

Recombinant bacteria capable of metabolizing sucrose are described. The recombinant bacteria comprise in their genome or on at least one recombinant construct: a nucleotide sequence from Bacillus licheniformis ATCC® 14580 encoding a polypeptide having sucrose transporter activity and a nucleotide sequence from Bacillus licheniformis ATCC® 14580 encoding a polypeptide having sucrose hydrolase activity. These nucleotide sequences are each operably linked to the same or a different promoter. Recombinant bacteria capable of metabolizing sucrose to produce glycerol and / or glycerol-derived products such as 1,3-propanediol and 3-hydroxypropionic acid are also described.

The present invention provides a method of producing optically active amino acids from 5-substituted hydantoin by isolating a hydantoinase gene and an N-carbamyl-L-amino acid hydrolase gene involved in an ability to convert 5-substituted hydantoin or N-carbamylamino acid into optically active amino acids from a microorganism of the genus Microbacterium having the above ability and by improving gene amplification and transcriptional and translational activities thereby preparing a recombinant wherein the amount of the desired enzymes produced is increased. The hydantoinase gene is, for example, a DNA encoding for a protein having a hydantoinase activity, which has the nucleotide sequence set forth in SEQ ID NO:1 in the Sequence. The N-carbamyl-L-amino acid hydrolase gene is, for example, a DNA encoding for a protein having an N-carbamyl-L-amino acid hydrolase activity, which has the nucleotide sequence set forth in SEQ ID NO:3 in the Sequence.

The invention discloses high-activity PET hydrolase mutants and application thereof. A protein provided by the invention is any one of the following 1)-11): 1) the protein shown in the specification is a protein with PET hydrolase activity obtained by mutating asparagine at the 204th site of wild PET hydrolase into alanine and mutating arginine at the 251th site into alanine and keeping amino acidresidues at other positions unchanged. According to the invention, original PET hydrolase (PETase) is mutated by utilizing a structure analysis and site-directed mutagenesis technology to obtain eight mutants, so that the problem that the original PET hydrolase is relatively low in activity is solved, the PET degradation activity of PETase is effectively improved, and the degradation effect of PETase is improved. PET is an insoluble high polymer which is difficult to degrade, the eight mutants greatly improve the degradation efficiency of PET plastic, and the industrial application prospect is promising.

A sugar ester derivative of the general formula (I) wherein G stands for a group of the formula (A) or a group of the formula (B) wherein lambda means 0 or 1; at least one of R1 and R2 means an ester residue of a saturated or unsaturated fatty acid having 1-30 carbon atoms and the other group means hydrogen atom or acetyl group, and R3 means a group of the formula (C) wherein X means a halogen atom, m means an integer of 0 to 4, Y means hydroxy group, an alkoxy group, a carboxyl group or sulfonic acid group, n means 0 or 1 and Z means nitro group or nitrovinyl group and its use. The sugar ester derivative of the general formula (I) is useful as a substrate for the measurement of lipase or esterase activities, and the reagents and the methods for measuring lipase or esterase activities which comprise said sugar ester derivative as the substrate are excellent in terms of reproducibility and sensitivity and enable the measurement of lipase or esterase activities in accordance with rate-assay method by an easy procedure.

The invention relates to polypeptide having pNPPC hydrolase activity and a coding gene, a preparation method and application thereof. The separated polypeptide is selected from: (1) amino acid sequence shown as SEQ ID NO:4, 6, 8 or 10; (2), SEQ ID NO:21 or segment obtained by truncating at least 15 amino acid residues from a terminal N thereof and / or at least 29 residues from a terminal C; (3), polypeptide derived from (1) or (2) by substituting, deleting or adding one or multiple amino acids in the amino acid sequence mentioned in (1) or (2) and maintaining pNPPC hydrolysis activity that SEQ ID NO: 4, 6, 8 or 10 or SEQ ID NO:21 or segment thereof; (4), polypeptide containing the amino acid sequence mentioned in (1), (2) or (3). The polypeptide like MPPX has performances like high substrate specifity and is quite suitable for application of enzyme in enzyme-linked immunosorbent assay.