Preparation method of porcine pepsin and gastric mucin

A technology of pepsin and peptin, which is applied in the field of preparation of pepsin and peptin, can solve the problems of only using hot air circulation, physical hazards of production workers, flammable and explosive organic solvents, etc., to protect the activity of pepsin , Shorten the production cycle and simplify the production process

Active Publication Date: 2015-04-01
四川德博尔制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The separation methods of pepsin include fractional salting-out separation, organic solvent separation and substrate affinity separation, etc. At present, the separation method of pig pepsin at home and abroad is generally to separate the crude extract prepared by the acid method through an organic solvent to obtain Mixed pepsin including pepsin A, pepsin B, pepsin C, pepsin D, mucous protein, peptin, etc. The organic solvent method generally uses water-soluble organic solvents such as acetone or ethanol, although organic solvents precipitate proteins. Well, the solvent is easy to remove, but the disadvantage is that it is easy to denature enzymes and active proteins, so the operation requires strict conditions. At the same time, the properties of the material before drying in the traditional process determine that only hot air circulation can be used, so the production capacity is also greatly limited. In addition, organic solvents are flammable and explosive, use a large amount, and cost high. Due to the certain toxicity of acetone, it will also cause certain harm to the health of production workers.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] (1) Preparation of crude extract: Take 1kg of pig stomach basal mucosa, add 270ml of 4% hydrochloric acid aqueous solution, stir evenly, heat up to 44-46°C in a water bath at 60-70°C, and adjust the pH to 2.2-2.4 after 30 minutes , acidify at constant temperature for 3 hours, filter with a 40-mesh nylon mesh, cool the filtered acidified gastric slurry to 0-3°C under stirring conditions, and obtain 1200ml of crudely extracted acidified gastric slurry;

[0024] (2) Separation: Add 4.8 L of phosphoric acid aqueous solution at 8 °C and pH 2.0 to 1.2 L of acidified gastric slurry, stir evenly, and after standing for 5 to 6 hours, filter the supernatant with 621 filter cloth until clarified to obtain pepsin The supernatant was collected for later use. The lower precipitate was centrifuged at a speed of 1500r / min. The supernatant was taken and filtered with a 621 filter cloth until clarified to obtain a pepsin supernatant, which was collected for later use. The centrifuged filt...

Embodiment 2

[0031] In the present embodiment, the preparation method of the crude extract is the same as in Example 1, and all the other preparation methods are as follows:

[0032] (1) Separation: Add 14.4 L of acetic acid aqueous solution at 10°C and pH 5.0 to 1.2 L of acidified gastric slurry, stir evenly, and after standing for 5 to 6 hours, filter the supernatant with 621 filter cloth until clarified to obtain pepsin The supernatant was collected for later use, and the lower precipitate was centrifuged at a speed of 1500r / min. The supernatant was taken and filtered with a 621 filter cloth until clarified to obtain a pepsin supernatant, which was collected for later use. The filter residue after centrifugation was 22.5g, and the temperature was 10°C, pH5 .0 90ml of acetic acid aqueous solution, stir evenly, centrifuge after standing for 20 minutes, take the supernatant and filter it with 621 filter cloth until the pepsin supernatant is clarified, collect it for later use, and filte...

Embodiment 3

[0039] In the present embodiment, the preparation method of the crude extract is the same as in Example 1, and all the other preparation methods are as follows:

[0040] (1) Separation: Add 9.6L of hydrochloric acid aqueous solution at 9°C and pH 3.0 to 1.2L of acidified gastric slurry, stir evenly, and after standing for 5-6 hours, filter the supernatant with 621 filter cloth until clarified to obtain pepsin The supernatant was collected for later use, and the lower sediment was centrifuged at a speed of 1500r / min. The supernatant was taken and filtered with a 621 filter cloth until clarified to obtain a pepsin supernatant, which was collected for later use. The filter residue after centrifugation was 50.5g, and the temperature was 9°C and pH3 .0 hydrochloric acid aqueous solution 101ml, stir evenly, centrifuge after standing for 20 minutes, take the supernatant and filter it with 621 filter cloth until the pepsin supernatant is clarified, collect for later use, and filter...

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PUM

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Abstract

The invention discloses a preparation method of porcine pepsin and gastric mucin, and the preparation method is designed for achieving the purpose of overcoming the defects that by using traditional pepsin preparation methods, pepsin is easily degenerated, organic solvents hurts the bodies of operators, and the cost is high, and the like. The method comprises the following steps: carrying out crude extraction on the mucosa of a basilar part of a porcine stomach by using a hydrochloric acid; then, by using the properties that pepsin is high in solubility and stable in enzyme activity in a weakly acidic solution, and gastric mucin is precipitated after meeting a weak acid, extracting and separating porcine pepsin and gastric mucin by using an acidic aqueous solution replacing acetone; and carrying out centrifugation, filtration, ultrafiltration, drying, and the like on the obtained product, so that porcine pepsin and gastric mucin are obtained. In case that the method is adopted for preparing porcine pepsin and gastric mucin, the activity of pepsin is protected, and meanwhile, because no organic solvent is used, the safety factor of operating is improved, and no harm is caused to the bodies of workers; and the cost is reduced, and the production capacity is increased, so that the method has broad market prospect.

Description

technical field [0001] The invention relates to a preparation method of pepsin and peptin, in particular to a method for preparing pepsin and peptin with pig gastric mucosa. Background technique [0002] Pepsin is extracted from the gastric mucosa of pigs. It is a digestive protease secreted by the gastric mucosal principal cells in the stomach. Its function is to decompose the protein in the food into small peptide fragments. At present, the medicinal gastric Protease is generally extracted from the gastric mucosa of pigs and used for indigestion. Gastrin is a mucin obtained by digesting and extracting the gastric mucosa of pigs. It has anti-pepsin and weak anti-acid effects. It can form a film in the stomach to cover the ulcer surface, reduce the stimulation of gastric acid, and is beneficial to the healing of the ulcer surface. The separation methods of pepsin include graded salting-out separation, organic solvent separation and substrate affinity separation, etc., but a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/64C07K14/47C07K1/14
CPCC07K14/4727C12N9/6481
Inventor 张革唐章勇余茂军周后杰陈红兵
Owner 四川德博尔制药有限公司
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