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39 results about "Oxalate decarboxylase" patented technology
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In enzymology, an oxalate decarboxylase (EC 4.1.1.2) is an enzyme that catalyzes the chemical reaction oxalate + H⁺ ⇌ formate + CO₂ Thus, the two substrates of this enzyme are oxalate and H⁺, whereas its two products are formate and CO₂. This enzyme belongs to the family of lyases, specifically the carboxy-lyases, which cleave carbon-carbon bonds. The systematic name of this enzyme class is oxalate carboxy-lyase (formate-forming).
Polynucleotides encoding oxalate decarboxylase from Aspergillus niger and methods of use
ActiveUS6929940B1Prevent oxalate accumulationPrevent encrustationFungiSugar derivativesNucleotideBacillus subtilis
The subject invention pertains to polynucleotides encoding the enzyme oxalate decarboxylase from the filamentous fungus Aspergillus niger and methods of use. The subject invention also pertains to methods of using the enzyme oxalate decarboxylase from Bacillus subtilis.
Owner:UNIV OF FLORIDA RES FOUNDATION INC +1
Preparation method of cross-linked oxalate decarboxylase aggregates (CLEAs)
InactiveCN102492683AHigh purityImprove separation efficiencyImmobilised enzymesMicroorganism based processesDiseaseCross-link
The invention provides a method for cross-linking and fixing oxalate decarboxylase by cross-linked oxalate decarboxylase aggregates (CLEAs), belonging to the technical field of biochemical engineering. The method comprises the following steps: induced expression of recombinant oxalate decarboxylase, separation and purification of oxalate decarboxylase, and preparation of CLEAs. The invention is characterized in that ethanol, acetone or ammonium sulfate are utilized to carry out precipitation separation on oxalate decarboxylase in a crude enzyme liquid; ethanol is utilized to prepare the oxalate decarboxylase aggregates; 0.01-0.15% glutaric dialdehyde is utilized to carry out cross-linking reaction for 0.5-3.5 hours under the conditions of adding 0.1-1.5mg / L BSA (bovine serum albumin), pH value 3-9 and 4 DEG C, thereby preparing the CLEAs; and the enzyme activity recycling rate of the cross-linked immobilized enzyme is up to more than 90%. The invention has the advantages of simple experimental operations and low cost; the obtained CLEAs have obviously higher stability than free oxalate decarboxylase; and thus, the invention can be used for developing enzyme preparations for oxalate calculus disease.
Owner:南宁奕德环境科技有限公司
Oxalate decarboxylase and recombinant expression method of oxalate decarboxylase
PendingCN105695441AOvercoming the inability to produce via E. coli expression systemsExpression realizationPeptide/protein ingredientsUrinary disorderEscherichia coliCompetent cell
The invention discloses an oxalate decarboxylase and a recombinant expression method of the oxalate decarboxylase. The oxalate decarboxylase is subjected to recombinant expression through an Escherichia coli expression system, and is stable and active when the pH value is less than 3.0; and compared with the primary zymoprotein sequence, a signal peptide segment is deleted in the protein sequence of the oxalate decarboxylase in the expression process. The recombinant expression method is suitable for oxalate decarboxylase which is still active when the pH value is lower than 3.0: the DNA segment of the optimized oxalate decarboxylase coding gene and an Escherichia coli expression vector are mixed according to the mole ratio of (3-9):1 to perform connection so as to establish the recombinant expression vector, and the recombinant expression vector is used for transforming DH5a competent cells. PCR (polymerase chain reaction) and sequencing verification are utilized to screen the successfully established recombinant expression vector transformed bacteria, the recombinant expression vector is extracted after culture and is used for transforming the molecular-chaperone-containing Escherichia coli expression strain, and 15-30-DEG C culture and protein expression induction are performed. The method enhances the expression efficiency, simplifies the production steps and lowers the production cost.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Method for preparing oxalic acid decarboxylase by fungus
The invention discloses a preparing method of oxalic acid decarboxylase with fungi, which comprises the following steps: (1) moist-heat-sterilizing organic acid or inorganic acid solution with high temperature at 121 deg. c; (2) setting the initial pH value of liquid medium of trametes versicolor (Trametes versicolor) or aspergillus niger (Aspergillus niger) and so on fungi at 5. 0-5. 5; setting the reacting temperature at 20-25 deg. c and air capacity at 20-30L / L . h; adding the acid solution at the third or fourth day with pH value at 0-3; centrifuging; collecting thallus cell; or filtering; getting thallus cell; eluting with o. 05-0. 2M and pH value at 3-3. 7 acetic acid buffer solution; collecting; freezing; grinding hypha; adding into acetic acid-sodium acetate buffer solution with 0. 05-0. 2M and pH value at 3-3. 7; grinding to pasty continually; centrifuging; getting oxalic acid decarboxylase liquid. This invention possesses low cost, which can be used to medical, food, beverage, agriculture and so on aspects.
Owner:DONGHUA UNIV
Recombinant expression plasmid vector and recombinant strain to be used in producing oxalate decarboxylase, and method of producing recombinant oxalate decarboxylase
It is intended to provide a means of highly producing oxalate decarboxylase originating in a microorganism. A recombinant expression plasmid vector, which contains a-amylase promoter of a microorganism belonging to the strain Bacillus and an oxalate decarboxylase gene originating in a microorganism that is provided under the control of the promoter, is constructed. A host bacterium is transformed by this vector to give an oxalate decarboxylase-producing bacterium. A recombinant oxalate decarboxylase is produced by culturing the producing bacterium as described above and then harvesting the oxalate decarboxylase thus produced.
Owner:AMANO ENZYME INC
Crystallized oxalate decarboxylase and methods of use
Oxalate decarboxylase crystals, including stabilized crystals, such as cross-linked crystals of oxalate decarboxylase, are disclosed. Methods to treat a disorder associated with elevated oxalate concentration using oxalate decarboxylase crystals are also disclosed. Additionally disclosed are methods of producing protein crystals.
Owner:ALTUS PHARMA INC
Preparation method of dry powder and fungus powder containing Tricholoma lobayense Heim oxalate decarboxylase
ActiveCN102697040AIncrease vitalityEfficient degradationPowder deliveryMicroorganism based processesIntestinal structureCalcium oxalate calculus
The invention discloses a preparation method of dry powder and fungus powder containing Tricholoma lobayense Heim oxalate decarboxylase and relates to the field of degradation of oxalate in stomachs and intestines of human bodies or animal bodies. The preparation method comprises the following steps of: using Tricholoma lobayense Heim or sibling fungi thereof as raw materials, culturing thalli by adopting a liquid fermentation method, regulating pH (potential of hydrogen) of fermentation solution to 2.0-5.0, inducing to produce oxalate decarboxylase, collection the fermentation solution and the thalli, crushing the thalli, conducting solid-liquid separation to respectively obtain liquid and solid containing the Tricholoma lobayense Heim oxalate decarboxylase, and respectively drying the obtained liquid and solid after separation to obtain the dry powder and the fungus powder containing the Tricholoma lobayense Heim oxalate decarboxylase. The dry powder and the fungus powder containing the Tricholoma lobayense Heim oxalate decarboxylase can be used for oxalic acid removal of food, beverage, feed and the like, can also be used as raw materials to prepare healthcare products or medicines, is suitable for oral taking, can effectively degrade oxalic acid and oxalate and can prevent or cure urine oxalic acid excess symptoms and calcium oxalate calculi.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Preparation method and application of oxalate decarboxylase
ActiveCN106939315AStrong secretory abilityHigh densityVectorsFermentationBiotechnologySequence signal
The invention discloses a preparation method of oxalate decarboxylase. The method comprises the following steps: constructing a recombinant expression plasmid vector comprising Bacillus-containing acid induced promoter, a secretion signal peptide, and a fungus-derived oxalate decarboxylase gene prepared under the control of the secretion signal peptide, transforming bacilli normally growing in a pH value being not more than 4.5 by the vector to prepare oxalate decarboxylase recombinant bacteria; and carrying out culture and acid feeding on the recombinant bacteria in a fermentation medium, inducing the recombinant bacteria, and carrying out simple purification to recover generated extracellular oxalate decarboxylase. The extracellular oxalate decarboxylase prepared through the method is used in diagnosis reagent raw materials, medicinal compositions, healthcare foods or formulated foods with special medical uses. The mutated acid induced promoter is adopted to substitute an original acid induced promoter in the recombinant expression vector, so the transformed bacilli have a high expression level.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Method for modifying oxalate decarboxylase with monomethoxypolyethylene glycol
InactiveCN103614359AImprove thermal stabilityImprove heat resistanceMicroorganism based processesEnzyme stabilisationMonomethoxypolyethylene glycolHeat resistance
The invention relates to a method for modifying oxalate decarboxylase, particularly a method for modifying oxalate decarboxylase with monomethoxypolyethylene glycol, belonging to the technical field of biochemical engineering. The method comprises the following steps: preparation of recombinant oxalate decarboxylase, activation of monomethoxypolyethylene glycol (mPEG), modification of oxalate decarboxylase and the like. The modified oxalate decarboxylase has higher heat stability, pH adaptability, heat resistance and trypsinase tolerance. The method solves the realistic problem that the oxalate decarboxylase in use can be influenced by digestion of gastric acid and proteinase and can be easily digested to lose activity at present. The method can be used for preparing an enzyme preparation for preventing and treating calculus caused by oxalate crystallization.
Owner:南宁奕德环境科技有限公司
Crystallized oxalate decarboxylase and methods of use
Oxalate decarboxylase crystals, including stabilized crystals, such as cross-linked crystals of oxalate decarboxylase, are disclosed. Methods to treat a disorder associated with elevated oxalate concentration using oxalate decarboxylase crystals are also disclosed. Additionally disclosed are methods of producing protein crystals.
Owner:味之素阿尔西亚有限公司
Composition for inhibiting or dissolving calcium oxalate stones as well as preparation method and application of composition
ActiveCN111419786AGrowth inhibitionNon-traumaticPeptide/protein ingredientsEnergy modified materialsOXALIC ACID DIHYDRATECalcium oxalate calculus
The invention relates to a composition for inhibiting or dissolving calcium oxalate stones as well as a preparation method and application of the composition. The composition comprises an immobilizedoxalate decarboxylase that can catalyze a decarboxylation reaction of oxalic acid or / and an oxalate, and the immobilized oxalate decarboxylase is prepared by immobilizing oxalate decarboxylase on a magnetic particle carrier. The composition provided by the invention has the function of inhibiting the growth of the calcium oxalate stones or dissolving the calcium oxalate stones, and has the advantages of mildness, good tolerance and high safety, and a patient has no trauma or pain when the composition is applied to treat kidney stones; and when the composition provided by the invention is applied to treat the calcium oxalate stones, the oxalate decarboxylase can be fixed in a renal pelvis by applying a magnetic field to continuously degrade oxalic acid in urine, so that the action time andefficiency of enzyme are increased, after the treatment is completed, the magnetic field is removed, and immobilized enzyme magnetic beads can be excreted with the urine; and after the composition isused in combination with drugs for regulating the pH of the urine, the effects of degrading the oxalic acid and dissolving the stones can be enhanced.
Owner:周定兰
Recombinant expression plasmid vector and recombinant strain to be used in producing oxalate decarboxylase, and method of producing recombinant oxalate decarboxylase
An object is to provide a means of highly producing an oxalate decarboxylase originating in a microorganism. A recombinant expression plasmid vector, which contains an α-amylase promoter belonging to the genus Bacillus and an oxalate decarboxylase gene originating in a microorganism that is provided under the control of the promoter, is constructed. A host bacterium is transformed with this vector to prepare an oxalate decarboxylase producing bacterium. A recombinant oxalate decarboxylase is produced by culturing the producing bacterium and then recovering the oxalate decarboxylase thus produced.
Owner:AMANO ENZYME INC
Glycosylated oxalate decarboxylase as well as preparation and application of glycosylated oxalate decarboxylase
PendingCN107868776AIncrease vitalityImprove degradation efficiencyFungiPeptide/protein ingredientsKidney stoneOxalate decarboxylase
The invention discloses glycosylated oxalate decarboxylase. The oxalate decarboxylase is derived from edible basidiomycete; the glycosylated oxalate decarboxylase has enzymatic activity in an environment that pH value is 1.5-7.5; and the activity is 10% greater than optimum activity in an environment that pH value is 1.5-2.0, which is 2U / mg greater than an activity unit. The invention also discloses a preparation method and an application of the glycosylated oxalate decarboxylase. The glycosylated oxalate decarboxylase provided by the invention can keep activity under a low-pH condition; and the glycosylated oxalate decarboxylase can be used for effectively preventing and treating kidney stone.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Method of culturing coriolus versicolor-induced oxalate decarboxylase through jerusalem artichoke carbon source
InactiveCN102533707ASimple methodEasy to operateMicroorganism based processesLyasesInorganic saltsMycelium
The invention relates to a method of culturing coriolus versicolor-induced oxalate decarboxylase through jerusalem artichoke carbon source, which comprises: (1) lixiviating jerusalem artichoke juice. (2) diluting the jerusalem artichoke juice to the total sugar content thereof is 7-100g / L, adding nitrogen source and inorganic salt to obtain fermentation medium, adjusting the pH value of fermentation medium to 4.0-5.0, sterilizating for preparation; (3) bringing the mycelium seed liquid of coriolus versicolor into fermentation medium for culturing two to six days; (4) when mycelium pellets are formed, adding oxalic acid with final concentration 1-100mM to induce oxalate decarboxylase, and obtaining mycelium after culturing 0.5-6 days; (5) breaking up the frozen mycelium, extracting mycelium fragments with acetate buffer, removing the residue, and obtaining oxalate decarboxylase crude enzyme. The invention has simple process, low cost, high efficiency, non-inhibition of substrate induced oxalate decarboxylase, and stable enzyme production.
Owner:DONGHUA UNIV
Method for promoting Eupergit C 250 to immobilize oxalate decarboxylase by using ammonium sulfate
The invention provides a method for promoting Eupergit C 250 to immobilize oxalate decarboxylase by using ammonium sulfate, belonging to the technical field of biochemical engineering. The method comprises the steps of inducible expression of recombinant oxalate decarboxylase, separation and purification of the oxalate decarboxylase by means of affinity chromatography, and immobilization of the oxalate decarboxylase. The method is characterized in that the Eupergit C 250 containing active epoxy groups is used as an immobilized vector, and 0.1-2mol / L neutral salt of the ammonium sulfate is added in the immobilizing process to promote the adsorption and covalency connection reaction of the vector to the oxalate decarboxylase. The method provided by the invention is simple to operate, and enzymatic activity recovery rate reaches more than 84%; and the used vector material is moderate in price and has a stable materialization performance, and the prepared immobilized enzyme has a better operating characteristic, is easily recovered and reused, and can be applied to oxalate degradation in food making or papermaking and other industrial processes.
Owner:GUANGXI UNIV FOR NATITIES
Recombinant plasmid for production of oxalate decarboxylase, system and method for expressing escherichia coli, and application
ActiveCN108977455AIncrease the concentration of manganese ionsOptimizing culture induction conditionsBacteriaPeptide/protein ingredientsBiotechnologySolubility
The invention discloses a recombinant plasmid for production of oxalate decarboxylase, a system and a method for expressing escherichia coli, and application. The recombinant plasmid for production ofoxalate decarboxylase comprises an oxalate decarboxylase gene, a molecular chaperone gene and a gene for regulating and controlling intracellular manganese ion concentration. The recombinant plasmidis guided into an escherichia coli expression bacterial strain and the escherichia coli with MntP gene deletion to respectively obtain oxalate decarboxylase with solubility expression and activity; the culture temperature and induction temperature of the recombinant bacterial strain are optimized; when the culture temperature is 37 DEG C, and the induction temperature is 25 DEG C, the expression amount of the oxalate decarboxylase is higher; by utilizing the two steps of organic solvent precipitation and ammonia sulfate precipitation to simply purify, the specific activity of the oxalate decarboxylase can reach 45.6U / mg, and the activity is higher than 95%. By adopting the technical scheme, the recombinant plasmid has the advantages that the production and purifying technologies are simple; the expression amount and specific activity of the oxalate decarboxylase are higher, the industrialized amplification is easy, the cost is low, and the recombinant plasmid is suitable for the industrialized production and application of the oxalate decarboxylase.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Preparation method of cross-linked oxalate decarboxylase aggregates (CLEAs)
InactiveCN102492683BHigh purityImprove separation efficiencyImmobilised enzymesMicroorganism based processesDiseaseCross-link
The invention provides a method for cross-linking and fixing oxalate decarboxylase by cross-linked oxalate decarboxylase aggregates (CLEAs), belonging to the technical field of biochemical engineering. The method comprises the following steps: induced expression of recombinant oxalate decarboxylase, separation and purification of oxalate decarboxylase, and preparation of CLEAs. The invention is characterized in that ethanol, acetone or ammonium sulfate are utilized to carry out precipitation separation on oxalate decarboxylase in a crude enzyme liquid; ethanol is utilized to prepare the oxalate decarboxylase aggregates; 0.01-0.15% glutaric dialdehyde is utilized to carry out cross-linking reaction for 0.5-3.5 hours under the conditions of adding 0.1-1.5mg / L BSA (bovine serum albumin), pH value 3-9 and 4 DEG C, thereby preparing the CLEAs; and the enzyme activity recycling rate of the cross-linked immobilized enzyme is up to more than 90%. The invention has the advantages of simple experimental operations and low cost; the obtained CLEAs have obviously higher stability than free oxalate decarboxylase; and thus, the invention can be used for developing enzyme preparations for oxalate calculus disease.
Owner:南宁奕德环境科技有限公司
Method for inducing oxalate decarboxylase by coriolus versicolor cultured by corn stalk carbon source
InactiveCN101735995ASimple methodEasy to operateMicroorganism based processesLyasesFermentationCulture mediums
The invention relates to a method for inducing oxalate decarboxylase by coriolus versicolor cultured by corn stalk carbon source, which comprises the following steps: (1) soaking crushed corn stalks in acid solution and hydrolyzing the crushed corn stalks at the temperature of 100 to 150 DEG C; (2) preparing a fermentation medium and adjusting the pH value to 4.0 to 5.0; (3) inoculating a mycelium into a liquid seed culture medium, when a white hyphal group is formed but does not float on a water surface, completely transferring the mycelium and the culture medium into a container containing sterile glass beads, vibrating and breaking the mixture, and inoculating the mixture into the fermentation medium in an inoculum size of 3 to 15 percent; and (4) culturing the mixture for 4 to 6 days, adding oxalic acid, taking out the mycelium after culturing the mixture for 0.5 to 6 days, and after freezing the mycelium with liquid nitrogen, grinding and crushing the mycelium, extracting hyphal fragments with acetic acid buffer solution, and centrifugating. The method of the invention is simple, easy in operation and low in cost; the preparation of a carbon source is strong in controllability; and the sugar content is stable.
Owner:DONGHUA UNIV
Method for culturing coriolus versicolor induced oxalate decarboxylase by using straw carbon source
The invention relates to a method for culturing coriolus versicolor induced oxalate decarboxylase by using a straw carbon source, which comprises the following steps of: (1) soaking crushed straws in acid solution, and hydrolyzing the crushed straws at the temperature of between 100 and 150 DEG C; (2) preparing a fermentation medium, and adjusting the pH value to between 4.0 and 5.0; (3) taking a mycelium, inoculating the mycelium into a liquid seed culture medium, when a white mycelium group is formed but is not emerged on the water surface, transferring the mycelium and the culture medium to a container containing sterile glass beads, vibrating the container to break the mixture, and inoculating the mycelium into the fermentation medium according to the inoculum size of 3 to 15 percent; and (4), after 4 to 6 days of culture, adding oxalic acid into the container, performing the culture for 0.5 to 6 days to obtain the mycelium, grinding and crushing the mycelium after the mycelium is chilled by liquid nitrogen, extracting mycelial fragments by using an acetate buffer, and centrifuging the mycelial fragments. The method is simple, is easy in operation and is low in cost, the preparation of the carbon source has strong controllability, and the sugar content is stable.
Owner:DONGHUA UNIV
Preparation of oxalate decarboxylase as well as product and application of oxalate decarboxylase
InactiveCN107868775AIncrease vitalityImprove efficiencyFungiPeptide/protein ingredientsFermentationOxalate decarboxylase
The invention discloses a preparation method of oxalate decarboxylase. According to the method, via biological fermentation technical means, acid-induced expression is carried out under a condition of0.001-50mM of manganese ions, so that the oxalate decarboxylase is produced. The invention also discloses a preparation prepared by the method and an application of the preparation. With the application of the method provided by the invention, the efficient preparation of the oxalate decarboxylase can be achieved.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Method for modifying oxalate decarboxylase by means of ethylenediaminetetraacetic dianhydride
InactiveCN106047847AMild conditionsEasy to operateEnzyme stabilisationCarbon-carbon lyasesHeat resistanceHeat stability
The invention discloses a method for modifying oxalate decarboxylase by means of ethylenediaminetetraacetic dianhydride. The method comprises the steps of dissolving ethylenediaminetetraacetic dianhydride and oxalate decarboxylase in phosphate buffer, and conducting stirring reaction for 1-24 h; dialyzing the reaction product after reaction ends, and then conducting concentration to obtain ethylenediaminetetraacetic dianhydride modified oxalate decarboxylase. Chemical modification of oxalate decarboxylase is directly conducted with ethylenediaminetetraacetic dianhydride, conditions are mild, operation is easy, and cost is low; the heat stability, heat resistance, pH adaptability and trypsin tolerance of modified oxalate decarboxylase are improved; the adsorption capacity of oxalate decarboxylase on calcium oxalate crystals is improved especially; the problem that oxalate decarboxylase can be affected by gastric acid and pepsin in use to lose activity easily is solved; the practical problem that zymoprotein in enzymic preparations is diluted easily to disappear in practical use is also solved.
Owner:南宁奕德环境科技有限公司
Preparation method of dry powder and fungus powder containing jinfu mushroom oxalate decarboxylase
ActiveCN102697040BIncrease vitalityEfficient degradationPowder deliveryMicroorganism based processesIntestinal structureTricholoma giganteum
The invention discloses a preparation method of dry powder and fungus powder containing Tricholoma lobayense Heim oxalate decarboxylase and relates to the field of degradation of oxalate in stomachs and intestines of human bodies or animal bodies. The preparation method comprises the following steps of: using Tricholoma lobayense Heim or sibling fungi thereof as raw materials, culturing thalli by adopting a liquid fermentation method, regulating pH (potential of hydrogen) of fermentation solution to 2.0-5.0, inducing to produce oxalate decarboxylase, collection the fermentation solution and the thalli, crushing the thalli, conducting solid-liquid separation to respectively obtain liquid and solid containing the Tricholoma lobayense Heim oxalate decarboxylase, and respectively drying the obtained liquid and solid after separation to obtain the dry powder and the fungus powder containing the Tricholoma lobayense Heim oxalate decarboxylase. The dry powder and the fungus powder containing the Tricholoma lobayense Heim oxalate decarboxylase can be used for oxalic acid removal of food, beverage, feed and the like, can also be used as raw materials to prepare healthcare products or medicines, is suitable for oral taking, can effectively degrade oxalic acid and oxalate and can prevent or cure urine oxalic acid excess symptoms and calcium oxalate calculi.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Method for determining concentration of oxalic acid by determining concentration of carbon dioxide through enzymatic method
InactiveCN101691601ASimple and fast operationLow costMicrobiological testing/measurementColor/spectral properties measurementsPhosphoenolpyruvate carboxylaseBuffer solution
The invention relates to a method for determining the concentration of oxalic acid by determining the concentration of carbon dioxide through an enzymatic method. The method comprises the following steps: (1) carrying out reaction of the mixture of oxalic acid, oxalate decarboxylase and acetate buffer solution for 30-60min at a temperature of 20-37 DEG C; (2) preparing Tris-HCI buffer solution containing phosphoenolpyruvate, phosphoenolpyruvate carboxylase, NADH, malic dehydrogenase, with a pH value of 7.3-8.5, incubating at a temperature of 37 DEG C and determining A1 at the position of 380nm; (3) adding the reaction solution prepared in the step (1) to the solution prepared in the step (2) for reaction for 5min at a temperature of 37 DEG C and determining A2 and A3 at the position of 380nm; and (4) calculating the decrement of NADH according to the formula: delta A 380nm = delta A to be determined - delta A blank, and calculating the concentration of the generated carbon dioxide according to a reaction principle so as to calculate the concentration of the oxalate decarboxylase. The method has simple operation, low cost and high sensitivity; the enzyme reagent adopted in the method is cheaper; and the method has similar accuracy compared with that of the existing kits for determining oxalic acid and has good practicability.
Owner:DONGHUA UNIV
A kind of preparation method and application of oxalate decarboxylase
The invention discloses a preparation method of oxalate decarboxylase. The method comprises the following steps: constructing a recombinant expression plasmid vector comprising Bacillus-containing acid induced promoter, a secretion signal peptide, and a fungus-derived oxalate decarboxylase gene prepared under the control of the secretion signal peptide, transforming bacilli normally growing in a pH value being not more than 4.5 by the vector to prepare oxalate decarboxylase recombinant bacteria; and carrying out culture and acid feeding on the recombinant bacteria in a fermentation medium, inducing the recombinant bacteria, and carrying out simple purification to recover generated extracellular oxalate decarboxylase. The extracellular oxalate decarboxylase prepared through the method is used in diagnosis reagent raw materials, medicinal compositions, healthcare foods or formulated foods with special medical uses. The mutated acid induced promoter is adopted to substitute an original acid induced promoter in the recombinant expression vector, so the transformed bacilli have a high expression level.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Fresh polygonatum odoratum freeze-dried decoction pieces and preparation method thereof
The invention discloses a fresh polygonatum odoratum freeze-dried decoction piece and a preparation method thereof, and relates to the field of traditional Chinese medicine, the fresh polygonatum odoratum is cleaned to remove impurities, the cleaned fresh polygonatum odoratum is cut into thick pieces, the thick pieces are placed in an acidic soak solution containing vegetable oil for ultrasonic impregnation, and the thick pieces are taken out and dried to obtain a first crude product; the first crude product is put into fermentation liquor containing oxalate decarboxylase to be fermented, and a second crude product is obtained after fermentation is finished; the second crude product is subjected to freeze drying under the vacuum condition, and a third crude product is prepared. According to the fresh polygonatum odoratum freeze-dried decoction piece, the content of calcium oxalate in the fresh polygonatum odoratum freeze-dried decoction piece is reduced, and meanwhile the dried decoction piece is not prone to embrittlement and easy to store.
Owner:江西纳弗堂制药有限公司
Method for culturing coriolus versicolor and inducing oxalate decarboxylase by rice straw carbon source
Owner:DONGHUA UNIV
Kit for measuring oxalic acid content of biological sample by enzyme coupling method and measuring method thereof
InactiveCN101788488AImprove accuracyImprove stabilityPreparing sample for investigationAnalysis by subjecting material to chemical reactionCouplingPre treatment
The invention relates to a kit for measuring the oxalic acid content of a biological sample by an enzyme coupling method. The kit comprises a sample pre-treatment kit and a sample reaction kit. The measuring method comprises the following steps of; pretreating the sample to be measured, and removing enzyme reaction inhibitors and interferents; adding oxalate decarboxylase into treated sample solution to perform a reaction at the temperature of between 15 to 16 DEG C for 20 to 60 minutes; adding solution of coenzyme I and deionized water, and measuring a light absorption value A1; adding formate dehydrogenlyase into the mixed solution to perform a reaction for 15 to 60 minutes, and measuring a light absorption value A2 of the reaction solution; and preparing a blank sample for comparison, and finally working out the oxalic acid content according to the measured absorption values. The measuring kit has the advantages of measuring samples from different sources, along with high sensitivity, high stability, convenient operation, low use cost and equipment cost, wide application range, high practicability and precision higher than other kits for measuring the oxalic acid content.
Owner:DONGHUA UNIV
A method for modifying oxalate decarboxylase with monomethoxypolyethylene glycol
InactiveCN103614359BImprove thermal stabilityImprove heat resistanceMicroorganism based processesEnzyme stabilisationMonomethoxypolyethylene glycolHeat resistance
The invention relates to a method for modifying oxalate decarboxylase, particularly a method for modifying oxalate decarboxylase with monomethoxypolyethylene glycol, belonging to the technical field of biochemical engineering. The method comprises the following steps: preparation of recombinant oxalate decarboxylase, activation of monomethoxypolyethylene glycol (mPEG), modification of oxalate decarboxylase and the like. The modified oxalate decarboxylase has higher heat stability, pH adaptability, heat resistance and trypsinase tolerance. The method solves the realistic problem that the oxalate decarboxylase in use can be influenced by digestion of gastric acid and proteinase and can be easily digested to lose activity at present. The method can be used for preparing an enzyme preparation for preventing and treating calculus caused by oxalate crystallization.
Owner:南宁奕德环境科技有限公司
A recombinant oxalate decarboxylase expressed in filamentous fungal host cells
ActiveCN108588060BIncrease secretionIncrease enzyme activityFungiMicroorganism based processesGenetic engineeringPromoter
The invention discloses a recombinant oxalate decarboxylase expressed by a filamentous fungal host cell, a construction method of the recombinant bacterial strain, a production method and application of the recombinant enzyme, and belongs to the technical field of genetic engineering. The recombinant filamentous fungal host cell comprises one or more copies of an oxalate decarboxylase expression cassette integrated in its genome, the oxalate decarboxylase expression cassette including a promoter, a signal peptide coding sequence, an oxalate decarboxylase coding gene and a terminator. The above-mentioned host cells can be constructed by random integration or site-specific integration. In addition, the present invention also optimizes the medium formulations for different recombinant filamentous fungal host cells. The production of oxalate decarboxylase, through the construction of expression cassettes, vector construction, host cell construction and the adjustment of the final culture positive formula, the final yield and enzyme activity are greatly improved, effectively solving the problem of oxalate decarboxylase in the prior art. Artificial production cannot be industrialized on a large scale.
Owner:WUHAN KANGFUDE BIOTECH CO LTD
Method for modifying oxalate decarboxylase with ethylenediaminetetraacetic acid diacid anhydride
InactiveCN106047847BMild conditionsEasy to operateEnzyme stabilisationCarbon-carbon lyasesP phosphateTrypsin
The invention discloses a method for modifying oxalate decarboxylase by means of ethylenediaminetetraacetic dianhydride. The method comprises the steps of dissolving ethylenediaminetetraacetic dianhydride and oxalate decarboxylase in phosphate buffer, and conducting stirring reaction for 1-24 h; dialyzing the reaction product after reaction ends, and then conducting concentration to obtain ethylenediaminetetraacetic dianhydride modified oxalate decarboxylase. Chemical modification of oxalate decarboxylase is directly conducted with ethylenediaminetetraacetic dianhydride, conditions are mild, operation is easy, and cost is low; the heat stability, heat resistance, pH adaptability and trypsin tolerance of modified oxalate decarboxylase are improved; the adsorption capacity of oxalate decarboxylase on calcium oxalate crystals is improved especially; the problem that oxalate decarboxylase can be affected by gastric acid and pepsin in use to lose activity easily is solved; the practical problem that zymoprotein in enzymic preparations is diluted easily to disappear in practical use is also solved.
Owner:南宁奕德环境科技有限公司
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