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Joint production process for producing alkaline phosphatase and heparin sodium with pig small intestine as raw material

A technology of pig small intestine and heparin sodium, applied in the field of biomedicine, can solve the problems of complex structure of heparin, unclear precise structure of heparin, abnormally complex structure of heparin, etc., and achieve the effects of optimizing enzymatic hydrolysis parameters, good hydrolysis effect and reducing production cost.

Active Publication Date: 2010-09-22
郑州弗博瑞生物技术有限公司
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Problems solved by technology

The structure of heparin is relatively complex. It is generally believed that it is composed of α-L-iduronic acid-2-sulfate, N-sulfo-α-D-glucosamine 6-sulfate, β-D-glucuronic acid and N-sulfo-α-D-glucosamine-6-sulfate uses glycosidic bonds to form "tetrasaccharides" as structural units, and then polymerizes "tetrasaccharides" into polysaccharides, which makes the entire structure of heparin extremely complicated. So far, the precise structure of heparin is unknown

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  • Joint production process for producing alkaline phosphatase and heparin sodium with pig small intestine as raw material
  • Joint production process for producing alkaline phosphatase and heparin sodium with pig small intestine as raw material

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Embodiment Construction

[0041] The present invention will be further described below in conjunction with the specific preparation methods and processes of each target object, but does not limit the protection scope of the present invention.

[0042] 1. Isolation and purification of alkaline phosphatase

[0043] (1) Clean the small intestine of the pig, cut its mucous membrane into pieces, add 0.05mol / L Tris-HCl buffer (pH7.5, containing 0.01mol / L Magnesium Chloride-0.01mol / L Sodium Chloride-0.01mol / L Zinc Chloride), fully homogenize in a beater, and then place the slurry at a low temperature of 4-10°C for more than 10h.

[0044] (2) Add pre-cooled n-butanol to the homogenate to make the concentration of n-butanol reach 20%-25% (V / V). After fully stirring, keep stirring in a water bath not higher than 37°C for 20-20- 30min, then place at room temperature for 8-10h.

[0045] (3) After pressing with a plate and frame filter press or centrifuging with a centrifuge (3000-5000r / m), the filtrate is colle...

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Abstract

The invention discloses a co-production production process of alkaline phosphatase and heparin sodium, which uses porcine small intestinal as the raw material. The process comprises the following steps: (1) the porcine small intestinal is refined and grinded; (2) 0.05mol / LTris-HCl buffer is added for extraction; (3)butanol precipitation; (4)pressure filtration and centrifugal separation; (5) the supernatant which is isolated from step(4) is precipitated by acetone; then the alkaline phosphatase is obtained through resolution ion exchange chromatography separation, concentration, gel chromatography separation, concentration and lyophilization; (6) the filter residue isolated from step(4) is processed with extraction, enzymatic hydrolysis, filtration, ion exchange resin absorption, impurities removal process by washing and elution, so that the heparin is obtained. The crude heparin sodium is obtained through alcohol precipitation, dehydration and drying; and the refined heparin sodium is obtained through hydrogen peroxide treatment, filtration, alcohol precipitation of the filtrate and acetone dehydration and drying. The process suits for the industrial production. The process can extract both alkaline phosphatase and refined heparin sodium. The invention is beneficial for the realization of the comprehensive utilization of the raw material. The invention reduces the production cost and decreases the environmental pollution of tail materials.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a joint production process for producing alkaline phosphatase and heparin sodium using pig small intestine as raw material, which can be widely used in biochemical reagents and biomedicine. Background technique [0002] (1) Alkaline phosphatase (Alkline phosphatase, EC 3.1.3.1, referred to as ALP or AKP) widely exists in the human body, animals, plants and microorganisms, and directly participates in physiological processes such as the transfer and metabolism of phosphate groups in organisms. The absorption and metabolism of calcium and phosphorus in the body and the maintenance of an appropriate ratio of calcium and phosphorus in the body play an important role. Alkaline phosphatase is a phosphomonoesterase with low substrate specificity, which can hydrolyze phosphate monoester bonds, but not phosphodiester bonds. It can act on a variety of substrates, such as β-glycerol phosphate, g...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/10C08B37/10
Inventor 王章存吴学军王斌
Owner 郑州弗博瑞生物技术有限公司
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