Method for preparing silkworm pupa protein ACE (angiotensin-I converting enzyme) inhibitory peptide by continuous enzyme membrane reaction, and product and application thereof

A technology of silkworm chrysalis protein and enzyme-membrane reaction, which is applied in the field of bioengineering, can solve the problems such as the preparation of silkworm chrysalis ACE inhibitory peptides, and achieve the effects of low price, realization of recycling, and prevention of activity reduction or loss

Inactive Publication Date: 2013-07-10
JIANGSU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there has been no report on the preparation of silkworm chrysalis ACE inhibitory peptides by continuous enzymatic membrane reaction

Method used

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  • Method for preparing silkworm pupa protein ACE (angiotensin-I converting enzyme) inhibitory peptide by continuous enzyme membrane reaction, and product and application thereof
  • Method for preparing silkworm pupa protein ACE (angiotensin-I converting enzyme) inhibitory peptide by continuous enzyme membrane reaction, and product and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Prepare the silkworm chrysalis protein into a solution with a substrate concentration of 0.5% (w / v), treat it in a 200W ultrasonic bath for 30 minutes, adjust its pH to 8.0 with NaOH solution, and add it to the reaction tank after heating to 40 °C. Add 2500U / g of alkaline protease (Alcalase), stir evenly, and carry out enzyme membrane reaction at 40°C. During the whole reaction process, the pH of the reaction solution is kept at 8.0 with NaOH solution, and the molecular weight cut-off of the ultrafiltration membrane is 3kDa. The pressure is 0.5kg / cm 2 , the peptide output rate was 6mL / min; the retained liquid during the reaction was returned to the reaction tank, and the permeate was collected. At the same time, the same concentration of silkworm chrysalis protein solution was added to the reaction tank according to the concentration of the product in the permeate. After the pH of the permeate is adjusted to neutral, use a vacuum concentrator to concentrate the peptide ...

Embodiment 2

[0029] Prepare the silkworm chrysalis protein into a solution with a substrate concentration of 2.0% (w / v), treat it in a 200W ultrasonic bath for 30 minutes, adjust its pH to 9.0 with NaOH solution, and add it to the reaction tank after heating to 50 °C. Add 3500U / g of alkaline protease (Alcalase) into the solution, stir evenly, and carry out enzyme membrane reaction at 50°C. During the whole reaction process, the pH of the reaction solution is kept at 9.0 with NaOH solution, and the molecular weight cut-off of the ultrafiltration membrane is 5kDa. The pressure is 0.7kg / cm 2 , the peptide output rate was 3mL / min; the retained liquid during the reaction was returned to the reaction tank, and the permeate was collected. At the same time, the same concentration of silkworm chrysalis protein solution was added to the reaction tank according to the concentration of the product in the permeate. After adjusting the pH of the permeate to neutral, use a vacuum concentrator to concentr...

Embodiment 3

[0031]Prepare silkworm chrysalis protein into a solution with a substrate concentration of 3.5% (w / v), treat it in a 200W ultrasonic bath for 30 minutes, adjust its pH to 9.5 with NaOH solution, and add it to the reaction tank after heating to 55 °C. Add 4500U / g of alkaline protease (Alcalase), stir well and then carry out enzyme membrane reaction at 55°C. During the whole reaction process, the pH of the reaction solution is kept at 9.5 with NaOH solution, and the molecular weight cut-off of the ultrafiltration membrane is 10kDa. The pressure is 1.0kg / cm 2 , the peptide output rate was 4mL / min; the retained liquid during the reaction was returned to the reaction tank, and the permeate was collected, and the same concentration of silkworm chrysalis protein solution was added to the reaction tank according to the concentration of the product in the permeate. After the pH of the permeate is adjusted to neutral, use a vacuum concentrator to concentrate the peptide content to about...

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Abstract

The invention relates to a method for preparing silkworm pupa protein ACE (angiotensin-I converting enzyme) inhibitory peptide by continuous enzyme membrane reaction. In the method, alkaline protease is taken as hydrolytic enzyme; and the silkworm pupa protein ACE inhibitory peptide is prepared by an enzyme membrane reactor. The preparation method comprises the steps of preparing solution from silkworm pupa protein and then reacting together with alkaline protease inside a reaction tank, carrying out real-time separation by an ultrafiltration membrane, returning trapped fluid to a reaction tank to react, and obtaining the powder silkworm pupa protein ACE inhibitory peptide from permeate liquid after vacuum concentration and spray drying, wherein IC50 is 0.10-0.50mlg / mL. The product disclosed by the invention is high in product yield, convenient to operate, low in production cost, and suitable for continuous production; and the obtained product accords with the requirements of functional food.

Description

Technical field: [0001] The invention belongs to the field of bioengineering, relates to a production method of functional food and biopharmaceuticals, in particular to a method for preparing silkworm chrysalis protein ACE inhibitory peptide by continuous enzyme membrane reaction, and its product and application. Background technique: [0002] Angiotensin-I converting enzyme (ACE) is a dipeptide carboxypeptidase that regulates human blood pressure. ACE controls the renin-angiotensin system (RAS) and kinin Liberase-kinin system (Kallikrein-kinin system, referred to as KKS) regulates human blood pressure. ACE inhibitory peptide is a biologically active peptide, which has the advantages of no toxic side effects and no effect on normal blood pressure. Long-term administration can achieve the purpose of prevention, control and auxiliary treatment of hypertension (Yu Z, Liu B, Zhao W, et al. Primary and secondary structure of novel ACE-inhibitory peptides from egg white protein [...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/34C07K2/00A61K38/01A61P9/12A23L1/29A23L33/00
Inventor 吴琼英贾俊强颜辉桂仲争徐金玲
Owner JIANGSU UNIV OF SCI & TECH
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