Preparation method of fish skin immunomodulatory peptides of Japanese croakers

A technology of immune regulation and yellow croaker, which is applied in the field of preparation of fish skin immunomodulatory peptides, can solve the problems of low utilization rate of fish skin protein, and achieve the effect of complete deodorization and debitterness, uniform peptide chain, and short degreasing time

Active Publication Date: 2018-11-02
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the low utilization rate of fish skin protein in the current enzymatic hydrolysis method to obtain polypeptides from fish skin, the purpose of this application is to provide a method for preparing the skin immunomodulatory peptide of Japanese yellow croaker. Fish skin leftovers are used as raw materials to prepare fish skin immunomodulatory peptides. The preparation process is gentle. The fish skin protein in fish skin is fully utilized and the activity is guaranteed. The obtained immunomodulatory peptides have high preparation efficiency and good activity.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A method for preparing the skin immunomodulatory peptide of Japanese yellow croaker, comprising the following steps:;

[0031] (1) Wash the fish skin with water and cut it into slices, then add water to the fish skin, and then add a mixture of lycopene and vitamin C with a weight of 0.2 wt% of the fish skin, the mass ratio of vitamin C and lycopene is 1:0.1, Then add lipase with an enzyme activity of 30,000U / g to degrease once at 30°C for 4 hours, rinse the fish skin until there is no lycopene color, then place it in 20wt% isopropanol solution for second degreasing for 4 hours, rinse the fish skin until there is no isopropanol smell and degrease stand-by;

[0032] (2) Soak the degreased fish skin in 2wt% sodium hydroxide low-concentration lye for 24 hours at a temperature of 25-30°C, then wash it with water until neutral, and then freeze it for later use;

[0033](3) Put the fish skin treated in step (2) in water, add acid solution to adjust the pH value to 2.0-4.6 / 2.8...

Embodiment 2

[0038] A method for preparing the skin immunomodulatory peptide of Japanese yellow croaker, the difference from Example 1 is:

[0039] The low concentration lye used in the step (2) is the sodium carbonate solution of mass concentration 2wt%;

[0040] Add acid solution in step (3) to adjust the pH value of the obtained soaking solution to 2.8, and the reductive amination reagent used is sodium borohydride;

[0041] The protein extraction enzyme used in step (4) is chymotrypsin.

[0042] The mass of fish skin immunomodulatory peptide was measured to be 2000~3500Da, with an average value of 3400±100Da.

Embodiment 3

[0044] A method for preparing the skin immunomodulatory peptide of Japanese yellow croaker, comprising the following steps:;

[0045] (1) Wash the fish skin with water and cut it into slices, then add water to the fish skin, add a mixture of lycopene and vitamin C with a weight of 0.4 wt% of the fish skin, the mass ratio of vitamin C and lycopene is 1:0.15, and then Add lipase with an enzyme activity of 40,000U / g to degrease once at 35°C for 3 hours, wash the fish skin until there is no lycopene color, then place it in 25wt% isopropanol solution for second degreasing for 3 hours, rinse the fish skin until there is no smell of isopropanol, and wait for the degreasing. use;

[0046] (2) Soak the degreased fish skin in a low-concentration sodium hydroxide lye with a mass concentration of 4wt% for 16 hours at a temperature of 27°C, then wash it with water until it is neutral, and then freeze it for later use;

[0047] (3) Put the fish skin treated in step (2) in water, add acid s...

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Abstract

The invention relates to the technical field of preparation of polypeptides of Japanese croakers and particularly relates to a preparation method of fish skin immunomodulatory peptides of Japanese croakers. The preparation method comprises the following steps: performing degreasing on fish skin by lipase combined with isopropanol to eliminate the bitter taste of immunomodulatory peptides; then, treating the fish skin through tartaric acid, a pulsed electric field and a reductive amination reagent to weaken the crosslinking degree among fish skin molecular proteins and enhance the swelling of the fish skin proteins; dissolving out the fish skin proteins in the fish skin as much as possible by virtue of the action of papain, chymotrypsin, trypsin, aspergillus serine protease or/and alkali protease and other protein extraction enzymes with weaker decomposition effect on the fish skin proteins at the temperature of 50-60 DEG C; and then, decomposing the fish skin proteins by virtue of thecombined action of neutral protease and neutral metal protease to obtain the fish skin immunomodulatory peptides. The utilization ratio of the fish skin proteins is high, the chain breaking decomposition uniformity is high, and the molecular weight of the obtained immunomodulatory peptides is 2000-3500Da.

Description

technical field [0001] The invention relates to the technical field of polypeptide preparation of Japanese croaker, in particular to a method for preparing the skin immunomodulatory peptide of Japanese croaker. Background technique [0002] Japanese croaker is a kind of fish belonging to the genus crocodile in the family Crowfish. It is distributed in North Korea, southern Japan and coastal areas of China. At present, it has been introduced and cultured in Zhoushan fishery in Zhejiang Province. It has rapid seedling growth and disease resistance. Strong ability, short growth cycle, high nutritional value, economic benefits and other advantages. Japanese croaker will produce scraps such as fish skin, fish bones, and fish viscera during the processing process. In order to further increase the economic value of Japanese croaker and rationally utilize resources, it is necessary to develop and utilize the approach of Japanese croaker scraps . [0003] Immunomodulatory peptides ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06
CPCC12P21/06
Inventor 余方苗董晓泽胡旭阳唐云平黄芳芳陈艳杨最素丁国芳
Owner ZHEJIANG OCEAN UNIV
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