Isolation Culture And Character Reseasch Of Metallothione-like Protein's Yeast

Metallothionein, MT for short, is a kind of low-molecular-weight, semi-hazy histidine-rich metal-binding protein . MT have oval-shaped molecules with molecular weight of 6500 Dalton, diameter 30-50A, two structural domains, each containing 7-12 molecules of metal atoms, has a special optical absorption. MT conformation stronger, with strong heat resistance.It shows good effect in removing the body of free radicals, delaying body aging; reducing the toxicity of heavy metals; involving in the metabolism of trace elements in the body; enhancing adaptability against all the body's bad status; Zinc storage; preventing cancer cells and etc. However, due to animal extracts, high cost and long cycle defects, the promotion application of MT has been limited. The purpose of this study is extracted MT from yeas by using short-cycle of bacteria and its low cost, and carry out experiments of the effectiveness of anti-oxidation, and eventually develop and utilize MT resources reasonably.Purpose:Isolated from the environment, have a filter with the ability of metallothionein microorganisms. To study issues affecting the microbial production of metallothionein an important factor, to draw the best process parameters. And metallothionein produced by oxidation of the anti-a comparative study.Method:Isolate and culture samples from Brewery wastewater and seperate four different thallus from it. 1) Process Zn-ion-induced on the four different thallus, and determine of the content of Zn-MT. 2) Analyze biochemical identification for the four dithallus, compare to "Yeast Identification Manual" and determine the species name.3) By choosing the concentration of Zn ions, temperature, PH, training time for the formal test, optimize the culture condition. 4) Carry out Fention reaction on MT, test Zn-MT's ability to eliminate hydroxy radical.Result:1) Comparing "Yeast Identification Manual", NO.1 is Candida Tropicalis, NO.2 is Rhodotorula Pallida, NO.3 is Saccharomyces Cerevisiae, NO.4 is Hansenula Anomala.2)after Zn inducement, the measured value of four different strains'MT were:0.885mg/g,0.673mg/g,0.591mg/g,0.721mg/g; 3) Through Biochemical identification and comparison of different MT, select Saccharomyces cerevisiae for culture conditions optimization, Saccharomyces cerevisiae metallothionein-like protein of high extraction, and no harm to human cells;the result indicates that when consistency of Zn is 30mg/L, temperature is 30°C, culture time is 24 hours and PH is 6.5, the contents of MT are 1.366mg/g, comparing the previous 0.885mg/g, the contents has increased 54.35%. 4) through determine the extraction of metallothionein-like protein, it shows that comparing the ability of ascorbic acid removing free radical, in the consistency between 0~50ug/mL, Zn-MT's clean effect is much stronger than ascorbic acid. In order to achieve the same clearance rate, the need of Zn-Mt is also higher than ascorbic acid.Conclusion:Isolated with the production capacity of metallothionein-like protein of the four yeast.Through the identification of yeast strains,The accuracy of the four strains were classified.W-1 is Saccharomyces cerevisiae,W-2 is Rhodotorula pallida,W-3 is Candida tropicalis,W-4 is Hansenula anomala.Through the optimization of culture conditions on yeast.The number of Metallothionein-like protein is increased from 0.885mg/g to 1.366mg/g,Increased about 54.35% percent.Comparing ascorbic acid's ability of eliminating hydroxy radical, metallothionein-like protein shows a good effect, which lay the foundation of developing and use of food yeast. Food yeast is rich in resources and has short training cycle. Metallothionein-like protein has good effect on eliminating hydroxyl radical, which is worth for further development and use.