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The Distribution of Selenium in Proteins of Saccaromyces Cerevisia and Analysis of Selenomethionine in Specific Protein

효모단백질내의 셀레늄 분포 및특정단백질내의 셀레노메티오닌 분석

  • 심희영 (단국대학교 자연과학대학 화학과) ;
  • 안상욱 (단국대학교 자연과학대학 화학과) ;
  • 안용현 (단국대학교 자연과학대학 화학과)
  • Published : 2003.08.20

Abstract

Selenized yeast (Se yeast) containing $0.1{\%}$(w/w) of selenium was obtained when the yeast was incubated at a selenium concentration of 1$1.14{\times}10_-3 M$ in rich medium. After washing several times, the inorganic selenium on the cell wall was confirmed with MBRT. There was no indication of inorganic selenium on the cell wall when the blue color in MBRT was stayed for 15 minutes. The selenized yeast was sonicated, then the selenium contained protein was obtained after salting out by ammonium sulfate at the concentration $80{\%}$ saturation. The seven protein bands were seperated by SDS-PAGE and the selenium concentration in protein was measured by ICP-AES. Analytical data showed that the large expressed protein band contained a relatively large amount of selenium. The proteins of the 47kDa was contained the concentrations of 69.5 ${\mu}$ Se/g of most many content. The protein (47 kDa) was seperated from PVDF membrane by tank-electroblotting. The isolated protein was hydrolyzed under acid condition and reacted with PITC. The derivatives of amino acids were analyzed by HPLC and compared with the data obtained from regular yeast. The resulting selenium-yeast was analyzed with the selenomethionine concentration of $2{\%}$ comparaed with general amino acids. The goal of this study is to analyze the selenium concentration in protein bands and measure the degree of biotransformation of selenomethionine in a specific protein.

Keywords

Selenium;Saccharomyces cerevisiae;Selenoamino Acids;Selenomethionine

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