The Distribution And Preparation Process Of Metallothionein In Visceral Organ Of Rabbit

Metallothionein(MT)is a kind of low molecular weight protein which widespread in organisms with rich cysteine and own metal binding ability.MT have a series of biological function,such as resistance to oxidation,Anti-aging,detoxification of heavy metals and so on.Rabbit is a kind of common animal,the purtenance of it account for a large proportion of the body,and the meat and the other organ are rich in nutrition,if not reasonable use,tremendous amounts of resources will be wasted.Studies have showed that rabbits viscera especially liver contains a large number of MT(2 mg/g),exploration is quite meaningful.This experiment is aimed at judged out the distribution of MT in rabbit organs,and research the preparation process of MT.This test using aqueous two-phase system(ATPS)to purify MT,so as to solve the problem of the high cost and the dangers of reagent toxic in the industrial prodution.In this paper,the conditions of the optimization test data are processed by Response surface method(RSM),optimum preparation process are obtained.Main research results are as follows:1.The distribution of MT in purtenance of rabbit:Liver > Kidney> Jejunum> Cecum> Spleen> Heart> Lung> Stomach wall> Blood> Bile.2.The extraction of liver MT: Mix up the Tris-HCl(0.02 mol/L pH 7.8)with the sample as a proportion of 1/6(m/v),the centrifugal condition is 4℃ 10000×g 30 min.heating in 80℃ water bath for 11 min,then get the MT crude production with 6000×g centrifugal.The best combinatorial optimization in ATPS are described as follows.The molecular weight of PEG 8000,the concentration of PEG 30%,the concentration of(NH4)2SO4 20%,pH 5,the sample amount 1 mL.MT extracted in rabbit kidney was proved to be Zn-MT.The content of the rabbit liver MT purified by AYPS was 1.98 mg/g,the purity reached 53.86%.3.The extraction of kidney MT: Mix up the Tris-HCl(0.02 mol/L pH 8.6)with the sample as a proportion of 1/6(m/v),the centrifugal condition is 4℃ 10000×g 30 min.heating in 82℃ water bath for 8 min,then get the MT crude production with 6000×g centrifugal.The best combinatorial optimization in ATPS are described as follows.The molecular weight of PEG 4000,the concentration of PEG 50%,the concentration of(NH4)2SO4 30%,pH 5,the sample amount 1.5 mL.MT extracted in rabbit kidney was proved to be Zn-MT.The content of the rabbit kidney MT purified by AYPS was 3.99 mg/g,the purity reached 90.07%.In summary,a certain amount of MT exist in rabbits offal,especially in the liver and kidney.This text proved that the purifucation effect of MT purified by ATPS in rabbit liver and kidney liver was significant.This shows that the ATPS is suitable for the extraction of MT.