Preparation method of silkworm pupa protein polypeptide

A technology of silkworm chrysalis protein polypeptide and silkworm chrysalis protein powder, which is applied in the field of preparation of silkworm chrysalis protein polypeptide, can solve the problems such as poor separation and purification effect of silkworm chrysalis polypeptide 2, and achieve good separation effect, high purity, fast and efficient separation effect

Active Publication Date: 2018-08-14
ZHEJIANG ESIGMA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the separation and purification effect of the silkworm chrysalis polypeptide 2 is not good

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The preparation method of silkworm chrysalis protein polypeptide comprises pretreatment of silkworm chrysalis powder, enzymatic hydrolysis of silkworm chrysalis protein powder, preparation of crude solution of silkworm chrysalis protein polypeptide, ion exchange resin chromatography, gel column chromatography, and high performance liquid chromatography purification. The specific steps are:

[0019] 1) Pretreatment of silkworm chrysalis powder: Add a mixture of n-hexane and petroleum ether with a volume ratio of 1:2 to silkworm chrysalis powder at a material-to-liquid ratio of 1:5 (g / mL), heat and reflux for 4 hours to obtain silkworm chrysalis protein, and set aside , due to the existence of oils and fats in silkworm chrysalis, the effect of enzymatic hydrolysis of silkworm chrysalis protein is greatly reduced, and at the same time, it will be oxidized in the enzymatic hydrolysis solution, resulting in bad flavor and deepening the color of the enzymatic hydrolysis solutio...

Embodiment 2

[0027] The preparation method of silkworm chrysalis protein polypeptide comprises pretreatment of silkworm chrysalis powder, enzymatic hydrolysis of silkworm chrysalis protein powder, preparation of crude solution of silkworm chrysalis protein polypeptide, ion exchange resin chromatography, gel column chromatography, and high performance liquid chromatography purification. The specific steps are:

[0028] 1) Pretreatment of silkworm chrysalis powder: Add a mixture of n-hexane and petroleum ether with a volume ratio of 1:1 to silkworm chrysalis powder at a material-to-liquid ratio of 1:8 (g / mL), heat and reflux for 5 hours to obtain silkworm chrysalis protein, and set aside ;

[0029] 2) Enzymatic hydrolysis of silkworm chrysalis protein powder: add distilled water to silkworm chrysalis protein powder according to the ratio of material to liquid 1:20 (g / mL), mix well, adjust the pH to 8.5, then add alkaline protease and neutral protease, and then add alkaline protease and neutra...

Embodiment 3

[0034] The preparation method of silkworm chrysalis protein polypeptide comprises pretreatment of silkworm chrysalis powder, enzymatic hydrolysis of silkworm chrysalis protein powder, preparation of crude solution of silkworm chrysalis protein polypeptide, ion exchange resin chromatography, gel column chromatography, and high performance liquid chromatography purification. The specific steps are:

[0035] 1) Pretreatment of silkworm chrysalis powder: Add a mixture of n-hexane and petroleum ether with a volume ratio of 1:1.5 to silkworm chrysalis powder at a material-to-liquid ratio of 1:6 (g / mL), heat and reflux for 4.5 hours to obtain silkworm chrysalis protein, spare;

[0036] 2) Enzymatic hydrolysis of silkworm chrysalis protein powder: add distilled water to silkworm chrysalis protein powder according to the ratio of material to liquid 1:22 (g / mL), mix well, adjust the pH to 8.0, then add alkaline protease and neutral protease, and then ℃ for 6 hours, then raise the temper...

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Abstract

The invention discloses a preparation method of silkworm pupa protein polypeptide. The preparation method comprises the following specific steps: adding a mixed solution of n-hexane and petroleum ether into silkworm pupa meal and carrying out heating reflux to obtain silkworm pupa protein; dissolving a silkworm pupa protein enzymatic hydrolysate into double distilled water, centrifuging to obtaina supernatant, namely silkworm pupa protein polypeptide raw liquor; adding distilled water into the silkworm pupa meal, uniformly mixing and adjusting a pH value; then adding alkaline protease and neutral protease, carrying out enzymolysis, enzyme deactivation and centrifuging, and concentrating the supernatant, namely the silkworm pupa protein polypeptide raw liquor; carrying out separation and purification on the silkworm pupa protein polypeptide raw liquor through ion-exchange resin chromatographic column, gel column chromatography and high performance liquid chromatography, thus obtainingthe silkworm pupa protein polypeptide. The preparation method of the silkworm pupa protein polypeptide, disclosed by the invention, has the beneficial effects of simplicity and feasibility; the protein polypeptide has the advantages of high yield, low loss, high purity and no possibility of destroying the activity; the obtained protein polypeptide has good inhibitory activity to ACE.

Description

technical field [0001] The invention relates to the technical field of polypeptide extraction, in particular to a preparation method of silkworm chrysalis protein polypeptide. Background technique [0002] China is a famous ancient silk country, and the "Silk Road" is famous for it. Sericulture has been an important part of China's agriculture since ancient times. There are records of silkworm raising and spinning in my country more than 7,000 years ago. At present, 80% of the world's silk production is produced in my country. Silkworm chrysalis protein is a high-quality protein resource. The protein content in dried silkworm chrysalis is as high as about 50%. It is rich in amino acids, of which 8 essential amino acids account for about 45% of the total, and the essential amino acids and non-essential amino acids are reasonable. , balanced, with a mass ratio of 0.73, in line with the reference protein model proposed by the World Health Organization / Food and Agriculture Orga...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/14C07K1/16C07K1/18
CPCC07K1/14C07K1/16C07K1/18C12P21/06
Inventor 陈贵才王贤玉徐天华吴中华张丽佳聂月美刘柳曹春华周晓红张小朋余有龙
Owner ZHEJIANG ESIGMA BIOTECH CO LTD
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