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Method for separating yak stomach lysozyme

A technology of lysozyme and yak, which is applied in the field of separation of yak stomach lysozyme, can solve the problems of no separation and purification of yak stomach lysozyme, and achieve the effect of good industrial application prospect, high yield and easy operation

Active Publication Date: 2015-02-25
SOUTHWEST UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is no literature report on the isolation and purification of yak stomach lysozyme

Method used

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  • Method for separating yak stomach lysozyme
  • Method for separating yak stomach lysozyme
  • Method for separating yak stomach lysozyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1 separation and purification method of the present invention

[0039] 1. Material collection

[0040] Maiwa yak abomasum was collected from Qingbaijiang slaughterhouse in Chengdu. After taking out the abomasum, wash it with normal saline. Then put it in an ice box, take it back to the laboratory, and store it at -20°C.

[0041] 2. Separation and purification

[0042] 2.1 Preparation of gastric mucosal homogenate

[0043] (1) Thaw the yak abomasum at 4°C. Then the gastric mucosa was torn off, and 7g of yak abomasum gastric mucosa was weighed, and 14ml of 10mM ammonium acetate (pH=6) was added thereto. Finally, the homogenate was processed with a tissue masher (the remaining gastric mucosa was stored at -80°C).

[0044] (2) Transfer the homogenate into a sterilized 50ml centrifuge tube, and centrifuge at 4°C (27,000×g, 15 minutes). The centrifuged supernatant was then transferred into a new 50ml centrifuge tube (stored at 4°C).

[0045] (3) Add 10mM am...

Embodiment 2

[0117] Example 2 Enzymatic properties of the yak abomasum lysozyme isolated and purified by the method of the present invention

[0118] Get the yak abomasum lysozyme prepared in Example 1 of the present invention, and detect its enzymatic properties:

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Abstract

The invention discloses a method for separating and purifying yak stomach lysozyme. The method comprises the following steps: (1) homogenizing to obtain homogenate; (2) carrying out coarse purification to obtain a coarse purified solution; (3) carrying out CM-Sepharose FF column chromatography to obtain a chromatographic solution; (4) carrying out Sephadex G-75 column chromatography to obtain a yak stomach lysozyme solution; and (5) and lyophilizing to obtain yak stomach lysozyme. By using the method disclosed by the invention, the pure yak stomach lysozyme can be separated from yak gastric mucosal tissue, has high enzyme activity, wide application range and antibacterial effect and can be used as an antibacterial agent. Meanwhile, the method is high in yield and simple and convenient to operate and has industrial application values.

Description

technical field [0001] The invention relates to a method for separating yak stomach lysozyme. Background technique [0002] Lysozyme is a basic protein, which is widely distributed in animals, plants and microorganisms, and is expressed in mammalian plasma, trachea, intestinal tract, stomach, kidney and liver tissue cells. Lysozyme lyses bacteria by disrupting the β-1,4 glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine in peptidoglycan, resulting in a structural change in the cell wall. Lysozyme can be used as a potential substitute for antibiotics in the fields of food, bioengineering, medical treatment, and livestock and poultry feed due to its antibacterial properties, non-toxicity, and side effects. From the perspective of livestock disease control, lysozyme is used as an additive in feed, which can effectively prevent livestock diarrhea, and at the same time provide nutrition and intestinal protection, thereby reducing the death of livestock caused b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/36A23K1/165A61K38/47A61P31/04A61P1/12
CPCA23K20/189A23K50/75A61K38/47C12N9/2462C12Y302/01017
Inventor 江明锋王永邓霄禹
Owner SOUTHWEST UNIVERSITY FOR NATIONALITIES
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