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Separation method of visceral protease in golden pomfret and protease as well as application

A separation method and protease technology, applied in the field of protease and its application, and the separation of protease from golden pomfret viscera, can solve the problems of low protease degradation ability, achieve the effects of simple production process, low raw material cost, and improved utilization rate

Active Publication Date: 2015-07-08
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a method for separating the protease from the viscera of golden pomfret, which solves the technical problem of low protease degradation ability for degrading gelatin in the prior art, and extracts a protease that efficiently degrades gelatin and is cheap from the viscera of golden pomfret for the first time

Method used

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  • Separation method of visceral protease in golden pomfret and protease as well as application
  • Separation method of visceral protease in golden pomfret and protease as well as application
  • Separation method of visceral protease in golden pomfret and protease as well as application

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Embodiment 1

[0060] Step 1: Wash the viscera of fresh golden pomfret and cut them into sections. Put the viscera in acetone at 4°C for 10 minutes to remove the fat according to the solid-liquid volume ratio of 1:2. After the defatting, use distilled water at 4°C Wash off the acetone on the viscera; then add the degreased viscera into 3 times the volume of Tris-HCl buffer solution with a pH of 7.5 at 4°C, fully break it with a beater, and then extract it at 4°C for 3 hours; finally, centrifuge at 4°C and 10,000 rpm for 15 minutes, and take the supernatant, which is the protease crude enzyme solution;

[0061] Step 2: Use ammonium sulfate to roughly separate the protease crude enzyme liquid to obtain a composite protease liquid. Firstly, ammonium sulfate is ground into a fine powder and added to the protease crude enzyme liquid to the following mass fractions: 25%, 35%, 45%, 55%, 65%, salting out to obtain crude composite protease liquid; then using casein as a standard substrate, using ultr...

Embodiment 2

[0064] Step 1: Wash the viscera of fresh golden pomfret and cut them into sections. Put the viscera in acetone at 4°C for 15 minutes to remove fat according to the solid-to-liquid volume ratio of 1:3, and use distilled water at 4°C after defatting Wash off the acetone on the viscera; then add the degreased viscera into 4 times the volume of Tris-HCl buffer solution with a pH of 7.5 at 4°C, fully break it with a beater, and then extract it at 4°C for 2 hours; finally, centrifuge at 4°C and 15,000 rpm for 20 minutes, and take the supernatant, which is the protease crude enzyme solution;

[0065] Step 2: Use ammonium sulfate to roughly separate the protease crude enzyme liquid to obtain a composite protease liquid. Firstly, ammonium sulfate is ground into a fine powder and added to the protease crude enzyme liquid to the following mass fractions: 25%, 35%, 45%, 55%, 65%, salting out to obtain crude composite protease liquid; then using casein as a standard substrate, using ultrav...

Embodiment 3

[0068]Step 1: Wash the viscera of fresh golden pomfret and cut them into sections. Put the viscera in acetone at 4°C for 20 minutes to remove fat according to the solid-to-liquid volume ratio of 1:2.5, and then use distilled water at 4°C after defatting Wash off the acetone on the viscera; then add the degreased viscera into 5 times the volume of Tris-HCl buffer solution with a pH of 7.5 at 4°C, fully break it with a beater, and then extract it at 4°C for 5 hours; finally, centrifuge at 4°C and 20,000 rpm for 30 minutes, and take the supernatant, which is the protease crude enzyme solution;

[0069] Step 2: Use ammonium sulfate to roughly separate the protease crude enzyme liquid to obtain a composite protease liquid. Firstly, ammonium sulfate is ground into a fine powder and added to the protease crude enzyme liquid to the following mass fractions: 25%, 35%, 45%, 55%, 65%, salting out to obtain crude composite protease liquid; then using casein as a standard substrate, using ...

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Abstract

The invention discloses a separation method of visceral protease in golden pomfret and protease as well as an application. The separation method comprises the steps of degreasing viscera of fresh golden pomfret, leaching and extracting, centrifuging to obtain protease crude enzyme, then carrying out crude separation on the protease crude enzyme by adopting ammonium sulfate to obtain composite protease liquid, finally separating the composite protease liquid obtained in the step two by adopting SephadexG-100 gel to obtain three different proteases which are recorded as a protease component I, a protease component 2 and a protease component 3. The separation method has the advantages that production residues in a golden pomfret processing process are fully utilized, the specific protease with strong gelatin degradation capacity is extracted from golden pomfret viscera, a new approach is opened for extraction of protease for resolving gelatin, the utilization rate of the golden pomfret is increased, and the high added value utilization of the golden pomfret is realized. In addition, the separation method is low in raw material cost and simple in a production technology.

Description

technical field [0001] The invention relates to the field of fish processing, in particular to a method for separating protease from viscera of golden pomfret, and also relates to the protease obtained by the above separation method and its application. Background technique [0002] Along with the large increase in demand of golden pomfret, a large amount of leftovers are produced in the processing process and are basically not developed and utilized, especially the viscera which accounts for 5% to 8% of the body weight of golden pomfret. This not only caused a huge waste of aquatic resources but also caused serious environmental pollution. Therefore, the high-value utilization of golden pomfret viscera is an urgent problem to be solved. [0003] Protease is a class of enzymes (enzymes) in organisms, they can decompose protein, widely exist in animal viscera, plant stems and leaves, fruits and microorganisms. Proteases are mainly extracted from plants, animals and microorg...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/64C12P21/06
CPCC12N9/6405C12P21/06
Inventor 申铉日肖鑫鑫
Owner HAINAN UNIVERSITY
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