35 results about "Atpase reaction" patented technology

The present invention provides an integrated microfluidic electrospray chip system and analytical method thereof, characterized in which the proteinase reaction, solid-phase extraction mechanism, electrophoresis and mass spectrometry are integrated into one system. This system allows continuous, fast and on-line detection and identification of a sample, where the sample is firstly hydrolyzed and desalted, and then introduced into the microfluidic electrospray chip to undergo electrophoresis. Finally, the separated sample is introduced into a mass spectrometer by means of electrospray for continuous detection and identification. This system applies mainly in the identification of biochemical substances, such as proteins.

The present invention provides an integrated microfluidic electrospray chip system and analytical method thereof, characterized in which the proteinase reaction, solid-phase extraction mechanism, electrophoresis and mass spectrometry are integrated into one system. This system allows continuous, fast and on-line detection and identification of a sample, where the sample is firstly hydrolyzed and desalted, and then introduced into the microfluidic electrospray chip to undergo electrophoresis. Finally, the separated sample is introduced into a mass spectrometer by means of electrospray for continuous detection and identification. This system applies mainly in the identification of biochemical substances, such as proteins.

The present invention relates to a method for preparing a palatability enhancer for use in pet foods of low, medium or high moisture content, comprising at least: a) providing a first stage reaction product obtained by: (i) reacting with at least one protease, in the absence of any added lipase, a substrate comprising proteinaceous and fatty materials, (ii) heat-inactivating said protease and filtrating the resulting digest product, b) optionally adding fat; c) emulsifying said first stage reaction product; d) reacting said emulsion with at least one lipase, in the absence of any added protease, so as to obtain a second stage reaction product; e) adding at least one reducing sugar and at least one nitrogen compound to said second stage reaction product and heating the resulting mixture.

An HGF precursor protein variant, in which a peptide structure comprises a sequence including a peptide chain X inserted between an α chain of HGF or a polypeptide where 1 to 20 amino-acid residues from the C-terminus of theα chain are deleted, and a β chain of HGF or a polypeptide where 1 to 20 amino-acid residues from the N-terminus of the β chain are deleted; wherein (i) the peptide chain X has an amino-acid sequence of at least two residues, (ii) the peptide chain X can be cleaved by a protease reaction or a chemical reaction, and (iii) a protein obtained by cleaving at least one site of the peptide chain X has HGF action.

A fast micro-protease reactor is prepared by taking multiple polymer monomers in capillaries, in-situ polymerizing in higher alcohol mixed solution to obtain continuous homogenous porous polymer substrate, activating while modifying for group and binding trypsase molecule on substrate to obtain the final product. It has fast high-activity protease molecule, better stability and less side reactions.

The invention relates to a pharmaceutical reagent, in particular to a reagent for determining the percentage of glycosylated hemoglobin. The reagent consists of the following substances: a dissolution buffering agent which is used for releasing hemoglobin from red blood cells in a blood sample, protease which can cut off a glycosylated valine or glycosylated polypeptide fragment of an N-terminal end of a beta chain of the glycosylated hemoglobin, an oxidant which can oxidize and eliminate interfering substances in the sample, an oxidant which can denature the hemoglobin and can accelerate the reaction rate of the protease, a reaction accelerator which can accelerate the reaction rate of the protease, fructosyl-amino acid oxidase undergoing oxidation-reduction reaction with the protease which can cut off the glycosylated valine or glycosylated polypeptide fragment of the N-terminal end of the beta chain of the glycosylated hemoglobin in the substance a, a color development agent which can undergo color development reaction with generated hydrogen peroxide, and peroxidase which can catalyze the hydrogen peroxide and the color development agent for the color development reaction. Compared with the prior art, the invention has the advantage that the reagent can directly determine the glycosylated hemoglobin.

The present invention relates to a method of producing keratin hydrolysate comprising the steps of: i) reacting keratin material with a protease; and ii) reacting keratin material with a chemical oxidant; wherein step ii occurs: a) after step i); b) during step i) when the selected protease hydrolyses under the pH conditions used for the chemical reaction and / or c) prior to step i) when the selected protease hydrolyses under the reaction conditions used for the chemical reaction; keratin hydrolysate so produced and uses thereof.

The present invention provides a protein comprising an amino acid sequence in which arginine at position 61 of a protein comprising the amino acid sequence represented by SEQ ID NO: 1 is substituted to an amino acid selected from the group consisting of glycine, alanine, valine, leucine, serine, threonine, proline, cysteine, methionine, asparagine, glutamine, and aspartic acid; and a method for measuring a glycated hemoglobin in a sample, wherein the method comprises reacting a glycated hemoglobin in a sample with a protease to produce a glycated hexapeptide, then reacting the produced glycated hexapeptide with the aforementioned protein, and measuring a substance produced or consumed by the reaction.

The invention relates to a preparation method of a hypotensive peptide of lactalbumin, belonging to the technical field of biology. The preparation method comprises the following steps of: maintaining 10min denaturation for lactalbumin suspension under the temperature of 85-90 DEG C, cooling to room temperature, adjusting the pH, then adding pepsin, reacting for 1-3 hours, deactivating enzyme, cooling, and centrifugating to take supernatant; adjusting the pH, adding trypsin and chymotrypsin, reacting for 4-6 hours, deactivating enzyme, cooling, and centrifugating to take supernatant; after vacuum concentration, freezing and drying to obtain a white or light-yellow crude product of the hypotensive peptide of the lactalbumin; and using DA201-C macroporous adsorption resin for adsorption, then carrying out gradient elution by alcohol, collecting and concentrating 75% of alcohol elution component, and utilizing glucan SephadexG-10 gel for chromatography separation to obtain the hypotensive peptide of the lactalbumin. The invention has the advantages that the preparation method is safe, has no toxic and side effects, has a simple process, and the industrial production is convenient; and the hypotensive peptide prepared by the invention can be easily absorbed by the human body and has high ACE (Angiotensin-converting Enzyme) inhibitory activity and has the characteristics of safety, no toxic and side effects, no action on normal blood pressure and the like, and can be used for preparing medicine and food for reducing blood pressure.

Described is a method for accurately and highly sensitively measuring glycated hemoglobin in a hemoglobin-containing sample without being influenced by hemoglobin. It is related to a method for measuring glycated hemoglobin in a hemoglobin-containing sample, comprising: reacting hemoglobin-containing sample with a protease in the presence of a surfactant; and then reacting the obtained reaction product with fructosyl peptide oxidase, wherein the latter reaction or both of the former reaction and the latter reaction are performed in the presence of a halogen oxide, and measuring the generated hydrogen peroxide. The method for measuring glycated hemoglobin in a hemoglobin-containing sample provided by the present invention is useful in, for example, the measurement of glycated hemoglobin useful in the diagnosis of diabetes mellitus.

The invention discloses an electrochemical sensing detection method of protein O-GlcNAc glycosyltransferase activity. The electrochemical sensing detection method comprises the following steps that a membrane of substrate polypeptide molecules of OGT is formed on the surface of an electrode; the obtained electrode fixed with the membrane of the polypeptide molecules on the surface and a glycosylation reaction fluid are incubated through OGT catalysis to obtain an electrode fixed with the membrane of the glycosylated polypeptide molecules on the surface; the electrode fixed with the membrane of the polypeptide molecules on the surface and the electrode fixed with the membrane of the glycosylated polypeptide molecules on the surface are sequentially subjected to the treatment in the steps 1-3 as follows: 1, the electrodes are put in a protease reaction fluid for hydrolysis; 2, the electrodes are cleaned with water; 3, a tyrosine residue serves as an electrochemical signal probe, and the electrodes perform electrochemical oxidation reaction; tyrosine residue electrocatalytic oxidized currents in the obtained systems are respectively determined, and OGT activity can be quantitively analyzed according to the obtained catalyzed and oxidized currents. By adopting the method, simple, quick and non-marked OGT activity detection is achieved through an electrochemical biosensor.

The invention discloses a method for extracting elastin peptide from bovine cartilage. The method comprises the following steps: grinding the bovine cartilage into powder; transferring the powder intoan extraction tank, adding purified water which is 3-4 times of the weight of the powder, heating a mixture to 95 DEG C, and performing stirring for 2 hours; cooling to 50-55 DEG C, adding compound protease, and after the reaction is finished, carrying out enzyme deactivation to obtain an enzyme shearing solution; cooling to 50-60 DEG C, and carrying out decolorization and deodorization treatment; separating a solute from a solvent by adopting a permeable membrane; separating organic small molecules with a molecular weight of 150-1000 Dal by adopting a nanofiltration membrane; and performingconcentration and spray drying. Protein peptides and non-peptide substances are different in thermal stability, the bovine cartilage powder is subjected to high-temperature extraction, an extracting solution is sheared through compound protease and then concentrated and filtered through different permeable membranes, small peptides with the molecular weight of 150-1000 Dal are obtained, the peptides with the molecular weight range account for 85% or above of peptides, and the absorption efficiency can be greatly improved.

The invention relates to the technical field of sludge treatment and especially relates to a method of enhancing sludge dehydration performance. The method particularly includes steps of: regulating pH value of wet sludge to 6.0-7.0; adding lysozyme to the wet sludge for a reaction, then adding protease for a reaction; after the enzyme pretreatment, dehydrating the wet sludge to prepare sludge filter cake. The method improves total dehydration performance of the sludge; after the dehydration through the method, the dehydrated sludge is reduced in influence on environment and ecologic system.

A sample preparation kit related to the present invention provides a significantly versatile analytical technique that is not affected by the diversity of antibodies, difference in species, matrix and the like. For preparing a sample to be used for detection of a monoclonal antibody through high-performance liquid chromatography-mass spectrometry (LC-MS), the kit includes a porous body for immobilizing a monoclonal antibody to be detected; nanoparticles with an immobilized protease; a reaction vessel for selectively digesting the monoclonal antibody by bringing the porous body and nanoparticles into contact; a buffer to be introduced into the reaction vessel along with the nanoparticles and porous body so that a protease reaction is carried out; and a filtration membrane to remove the porous body and nanoparticles after the proteolysis so as to extract the reaction product and the buffer.

The invention discloses a waste bacteria treatment method and a waste bacteria recycling method in waste bacterial production. The waste bacteria treatment method comprises: a, adjusting the pH valueof a spiromycin production fermentation broth to 4.0-6.5, and filtering by using a ceramic membrane to obtain a concentrated liquid retained by the ceramic membrane; b, adding lysozyme to the concentrated liquid to obtain a lysozyme reaction solution; c, adding protease to the lysozyme reaction solution to obtain a protease reaction solution; and d, carrying out spray drying on the protease reaction solution to obtain waste bacteria powder. The waste bacteria recycling method comprises: replacing the organic nitrogen source in fermentation production of spiramycin with the obtained waste bacteria powder. With the methods of the present invention, the waste bacteria can achieve the fermentation raw material condition, and can be reused for the production of the original product so as to avoid the external flow of the waste bacteria.

The invention relates to the enzyme catalysis / enzyme inhibition biotechnological field, in particular to a test method for activity of a trypsin inhibitor. The particular test process is mainly that: a trypsin inhibitor extract and a trypsin action substrate are respectively added into a trypsin reaction system; after reaction for 10 to 15 minutes, reaction stopping solution is added, and then absorbance of the reaction stopping solution in the wavelength of 410 nautical miles is measured; a curve is drafted according to addition and the absorbance of the inhibitors; the activity of the trypsin inhibitor is determined; wherein, the trypsin action substrate is BAPNA solution and the addition is 0.5 to 1.0 milliliters per milliliter trypsin reaction system. The invention has the advantages of simple instruments used, convenient operation and high reaction sensitivity, and is suitable for testing activity of various trypsin inhibitors.

Provided are fish meal proteolytic products, its preparation process and application, wherein the preparing process comprises, charging deionized water into fish meal, stirring in a container, charging ammonia to adjust the pH of the solution, charging basified protease for reaction, then charging ammonia to maintain the pH of the reaction system, water-bathing the reaction solution, removing enzyme, centrifuging, freeze drying the supernatant fluid into dried powder, finally freezing for later use.

The present invention relates to a method of producing keratin hydrolysate comprising the steps of: i) reacting keratin material with a protease; and ii) reacting keratin material with a chemical oxidant; wherein step ii occurs: a) after step i); b) during step i) when the selected protease hydrolyses under the pH conditions used for the chemical reaction and / or c) prior to step i) when the selected protease hydrolyses under the reaction conditions used for the chemical reaction; keratin hydrolysate so produced and uses thereof.

The invention discloses a preparation method and application of a polypeptide-containing sheep placenta zymolyte, and belongs to the technical field of enzyme engineering. The method mainly comprises the following steps: (1) taking sheep embryo homogenate or dry powder, adding water and conducting stirring; (2) conducting heating to 50-60 DEG C, adjusting the pH value to 8-9, adding alkaline protease to react for 0.5-12 hours, and then adding flavourzyme to react for 0.5-12 hours; and (3) heating the enzymatic hydrolysate obtained in the step (2) to 80-100 DEG C, conducting reacting for 25-45 minutes, conducting filtering, and drying the filtrate, thereby obtaining the product. The method disclosed by the invention is simple in process, low in raw material consumption and suitable for industrial production of the sheep placenta zymolyte; and the product obtained by the invention has good functions of improving sleep, improving climacteric syndrome and the like, and has a good market prospect.

The invention discloses a process for preparing a heavy metal adsorbent by waste plants. The specific process includes the steps: firstly, cleaning an appropriate quantity of Chinese yam residues by tap water and distilled water and drying the Chinese yam residues to reach constant weight; secondly, grinding the Chinese yam residues into powder and sieving the powder; thirdly, performing reaction of the sieved powder, amylase and protease for 1-2 hours under a certain condition, adding reactants into NaOH solution, performing reaction for 30-60 minutes, neutralizing the reactants with dilute H2SO4 solution, filtering and drying the reactants, mixing the reactants with urea according to a certain ratio, adding an appropriate amount of deionized water, performing reaction for 5-10 hours under the condition of 50 DEG C and then filtering the solution; finally, cleaning the reactants by the deionized water and drying the reactants to obtain the heavy metal adsorbent.

The present invention provides a protein comprising an amino acid sequence in which arginine at position 61 of a protein comprising the amino acid sequence represented by SEQ ID NO: 1 is substituted to an amino acid selected from the group consisting of glycine, alanine, valine, leucine, serine, threonine, proline, cysteine, methionine, asparagine, glutamine, and aspartic acid; and a method for measuring a glycated hemoglobin in a sample, wherein the method comprises reacting a glycated hemoglobin in a sample with a protease to produce a glycated hexapeptide, then reacting the produced glycated hexapeptide with the aforementioned protein, and measuring a substance produced or consumed by the reaction.

The invention discloses a histological method for distinguishing three kinds of carp muscle cells, wherein the method comprises the specific steps: freezing a treated sample with isoamyl and liquid nitrogen; freezing the sample at low temperature, slicing, and drying at room temperature; putting the slice into a constant-temperature pretreatment liquid, treating, and then cleaning with a cleaningliquid; adding a cleaning liquid into the slice, pouring out after the temperature is constant, adding an ATPase reaction liquid with the same temperature, carrying out a reaction, and cleaning and impregnating with distilled water; soaking the slice with a CaCl2 solution, a CoCl2 solution and an ammonium sulfide solution successively, and cleaning and impregnating with distilled water after the soaking is finished; absorbing excess moisture of the slice, covering with cover glass, sealing the slice, observing with an optical microscope, and thus distinguishing the three kinds of carp muscle cells. Boundary lines of red meat cells, pink meat cells and white meat cells in microscope images obtained by the distinguishing method are obvious, and the distribution of the three kinds of carp muscle cells can be clearly distinguished.

The invention relates to a pharmaceutical reagent, in particular to a reagent for determining the percentage of glycosylated hemoglobin. The reagent consists of the following substances: a dissolution buffering agent which is used for releasing hemoglobin from red blood cells in a blood sample, protease which can cut off a glycosylated valine or glycosylated polypeptide fragment of an N-terminal end of a beta chain of the glycosylated hemoglobin, an oxidant which can oxidize and eliminate interfering substances in the sample, an oxidant which can denature the hemoglobin and can accelerate the reaction rate of the protease, a reaction accelerator which can accelerate the reaction rate of the protease, fructosyl-amino acid oxidase undergoing oxidation-reduction reaction with the protease which can cut off the glycosylated valine or glycosylated polypeptide fragment of the N-terminal end of the beta chain of the glycosylated hemoglobin in the substance a, a color development agent which can undergo color development reaction with generated hydrogen peroxide, and peroxidase which can catalyze the hydrogen peroxide and the color development agent for the color development reaction. Compared with the prior art, the invention has the advantage that the reagent can directly determine the glycosylated hemoglobin.

The invention discloses a novel preparation process of a heavy metal absorbent. The preparation process of the heavy metal absorbent comprises steps as follows: waste potato dregs are taken as raw materials, cleaned, dried and ground into powder and then react with amylase and protease; a product then reacts with a NaOH solution and is subjected to a neutralization reaction with a dilute H2SO4 solution; then filtering and drying in the air are performed, dried powder is mixed with urea, and a proper amount of deionized water is added; filtering and cleaning are performed after the reaction, drying is performed finally, and the heavy metal absorbent can be prepared. The preparation process is simple and low in cost and has better application prospect.

The invention relates to the technical field of molecular biologics and discloses a high-flux screening method for a novel broad-spectrum lysozyme. The method comprises the following steps: A, performing secretory expression on a gene mutation library of a target lysozyme in eukaryotic cells so as to obtain fermentation liquids with different active target lysozymes; B, respectively adding the fermentation liquids with different active target lysozymes into a reaction system for reaction, and screening a lysozyme with high sterilization activity upon Gram negative bacteria, wherein the reaction system comprises Gram negative bacteria with intracellular expression of 'recognition site of donor fluorescin-protease-receptor fluorescin', protease and a protease reaction buffer solution. By adopting the method disclosed by the invention, through cascading of FRET (Fluorescence Resonance Energy Transfer) fluorescin pairs with locus specific protease, the detection sensitivity is remarkably improved; moreover, by adopting the method, high-flux detection on lysozymes can be achieved through a fluorescence microplate reader and a 96-microplate.

A process for preparing the high-purity antimer, L-p-hydroxyphenyl glycine includes such steps as sequentially adding ionic liquid BMIM.BF4, phosphoric acid buffer, methyl p-hydroxyphenylglycenate, and papain, reacting, ultrafilter to remove suspended particles and enzyme, and separating to obtain L-p-hydroxyphenyl glycine.