Method of extracting metallothionein with fish processing offcuts as raw material

Abstract

The invention discloses a method of extracting metallothionein with fish processing offcuts as a raw material. The method includes the steps of: 1) washing the fish processing offcuts as the raw material, and grinding the offcuts into pulp; 2) adding a Tris-HCl buffer solution, of which the pH is 7-8, in two times by weight of the raw material, and stirring the mixture for 30 s on a magnetic stirring apparatus; 3) adding a chloroform-ethanol mixed liquid according to the volume ratio of the Tris-HCl buffer solution to the chloroform-ethanol mixed liquid being 1:1-1.5, wherein the volume ratio of chloroform to ethanol is 0.08:1, and stirring the mixture for 4-6 min; 4) centrifuging the mixture at 5000-6000 r / min for 25 min at 4 DEG C to obtain a supernatant; 5) heating the supernatant to 75 DEG C for 3-5 min; 6) cooling the supernatant to 0 DEG C and filtering the supernatant through gauze in 8 layers, and allowing a filtrate to stand for 1.5-2 h; 7) centrifuging the filtrate at 8000-10000 r / min for 25 min at 4 DEG C to obtain a supernatant being a homogenized liquid; and 8) freeze-drying the homogenized liquid at -35 DEG C to prepare the product. On the basis that the Tris-HCl buffer solution can dissolve proteins, by regulating the pH value of the Tris-HCl buffer solution, the metallothionein in the fish processing offcuts can be extracted through centrifugal separation and freeze drying. The method is low in cost, is free of environment-pollution, is convenient to carry out and is controllable in process.

Description

technical field [0001] The invention relates to a method for extracting metallothionein by using leftovers after processing fish as raw materials, and belongs to the field of marine product processing. Background technique [0002] Metallothionein is a metal-binding protein produced by microorganisms, animals and plants. It is a short peptide rich in cysteine ​​and has a high affinity for various heavy metals. The important physiological function of metallothionein is that it is the most effective free radical scavenger known so far, its ability to scavenge free radicals is about five thousand times that of superoxide dismutase sod, and it has a strong antioxidant capacity active. The research on metallothionein mainly focuses on extraction, detection and application. At present, animal liver is mostly used as the raw material for extraction. my country is a large ocean country with rich marine resources. Many marine organisms contain metallothionein, especially in the lef...

AI Technical Summary

Problems solved by technology

my country is a large ocean country with rich marine resources. Many marine organisms contain metallothionein, especially in the leftovers after fish processing, which include fish heads, fish scales, and fish viscera. , fish bones and fish oil account for 30% to 50% of processed fish. If not treated effectively, it will not only pollute the environment, but also cause a lot of waste of resources