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

A technology of heparin sodium and phosphatase, applied in the field of biomedicine, can solve the problems of unclear precise structure of heparin, complex heparin structure, abnormally complex heparin structure, etc., and achieves the effects of good hydrolysis effect, optimization of enzymolysis parameters and reduction of production cost.

Active Publication Date: 2008-05-21
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

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  • Joint production process joint production 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

A co-production process for co-producing alkaline phosphatase and heparin sodium using pig small intestine as raw material, comprising the following steps: ① beating and crushing pig small intestine; ② adding 0.05mol/L Tris-HCl buffer solution for extraction; ③ n-butyl Alcohol precipitation; ④ pressure filtration or centrifugation; ⑤ The supernatant separated in step ④ is precipitated with acetone, and then separated by reconstitution ion exchange chromatography, concentrated, gel chromatographic separation, concentrated, and freeze-dried to obtain alkaline Phosphatase; ⑥ The filter residue separated in step ④ is extracted, enzymatically hydrolyzed, filtered, ion exchange resin adsorbed, washed to remove impurities, eluted to obtain heparin, and then subjected to alcohol precipitation, dehydration and drying to obtain crude heparin sodium, treated with hydrogen peroxide, filtered, The filtrate was precipitated with ethanol, dehydrated and dried with acetone to obtain fine heparin sodium. The process is suitable for industrial production, can not only extract alkaline phosphatase, but also can extract refined heparin sodium, which is beneficial to realize the comprehensive utilization of raw materials, reduce production costs, and reduce environmental pollution caused by leftovers.

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...

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Application Information

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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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