Separation and purification method of silkworm pupa protein polypeptide

A technology for the separation and purification of silkworm chrysalis protein polypeptides, which is applied in the field of polypeptide extraction, can solve the problems of poor separation and purification effect of silkworm chrysalis polypeptide 2, and achieve good separation effects, fast and efficient separation, and high resolution effects

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

[0017] The method for separating and purifying silkworm chrysalis protein polypeptides comprises preparation of silkworm chrysalis protein polypeptide crude liquid, ion exchange resin chromatography, gel column chromatography, and high performance liquid chromatography purification, and its specific steps are:

[0018] 1) Preparation of silkworm chrysalis protein polypeptide crude solution: Dissolve the silkworm chrysalis protein enzymatic hydrolyzate in double distilled water according to the ratio of material to liquid 1:10 (g / mL), and then centrifuge in a centrifuge at a temperature of 5 °C and a speed of 8000 rpm for 10 min , the supernatant is the crude silkworm chrysalis protein polypeptide liquid, which is ready for use. The components in the silkworm chrysalis protein crude polypeptide liquid are relatively complex, because their molecular weight and amino acid sequence are different, so they have different physical properties and biological activities. For more in-depth...

Embodiment 2

[0025] The method for separating and purifying silkworm chrysalis protein polypeptides comprises preparation of silkworm chrysalis protein polypeptide crude liquid, ion exchange resin chromatography, gel column chromatography, and high performance liquid chromatography purification, and its specific steps are:

[0026] 1) Preparation of silkworm chrysalis protein polypeptide crude liquid: according to the material-to-liquid ratio of 1:20 (g / mL), the enzymatic hydrolyzate of silkworm chrysalis protein was dissolved in double distilled water, and then centrifuged in a centrifuge at a temperature of 1°C and a speed of 10,000 rpm for 8 minutes , the supernatant is the crude silkworm chrysalis protein polypeptide liquid, which is set aside;

[0027] 2) Ion-exchange resin chromatography: The silkworm chrysalis protein peptide liquid was prepared into a solution with a concentration of 55 mg / mL with double distilled water, and then centrifuged in a centrifuge at a temperature of 1°C a...

Embodiment 3

[0033] The method for separating and purifying silkworm chrysalis protein polypeptides comprises preparation of silkworm chrysalis protein polypeptide crude liquid, ion exchange resin chromatography, gel column chromatography, and high performance liquid chromatography purification, and its specific steps are:

[0034] 1) Preparation of silkworm chrysalis protein polypeptide crude solution: Dissolve silkworm chrysalis protein enzymatic hydrolyzate in double distilled water according to the ratio of material to liquid 1:15 (g / mL), and then centrifuge in a centrifuge at 4°C and 9000rpm for 9min , the supernatant is the crude silkworm chrysalis protein polypeptide liquid, which is set aside;

[0035] 2) Ion-exchange resin chromatography: the silkworm chrysalis protein peptide solution was prepared with double distilled water to a concentration of 50mg / mL, and then centrifuged in a centrifuge at a temperature of 4°C and a speed of 9000rpm for 9min to remove insoluble impurities, an...

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Abstract

The invention discloses a separation and purification method of silkworm pupa protein polypeptide. The separation and purification method comprises the following specific steps: dissolving a silkwormpupa protein enzymatic hydrolysate into double distilled water and centrifuging to obtain a supernatant, namely silkworm pupa protein polypeptide raw liquid; dissolving the silkworm pupa protein polypeptide liquid into the double distilled water, centrifuging, adding a supernatant into an ion exchange resin chromatography column, eluting and collecting protein polypeptide which has optimum inhibitory activity on ACE, namely an ion exchange chromatography enzymatic hydrolysate; dissolving the ion exchange chromatography enzymatic hydrolysate into the double distilled water, centrifuging, carrying out sephadex column chromatographic separation on the supernatant, eluting with the double distilled water and collecting elution components according to an absorbance curve at 280nm; dissolving the gel chromatography enzymatic hydrolysate into the double distilled water and purifying by using high performance liquid chromatography, thus obtaining the silkworm pupa protein polypeptide. The separation and purification method of the silkworm pupa protein polypeptide, disclosed by the invention, has the beneficial effects that the separation method is simple and the protein polypeptide can beseparated quickly and efficiently; 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 on the ACE.

Description

technical field [0001] The invention relates to the technical field of polypeptide extraction, in particular to a method for separating and purifying silkworm chrysalis protein polypeptides. 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 A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14C07K1/16C07K1/18
CPCC07K1/14C07K1/16C07K1/18
Inventor 陈贵才王贤玉张小朋吴中华聂月美刘柳何奇雷程文虹徐天华严发杰余有龙
Owner ZHEJIANG ESIGMA BIOTECH CO LTD
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