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題名 | Enhancing the Production of γ-Glutamylcysteine by a Mutant Strain Derived from Saccharomyces Cerevisiae BCRC 21727=提高酵母BCRC 21727衍生之變異株中γ-麩氨半胱氨酸之產能 |
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作者 | 黃進發; 林欣儀; 楊芳鏘; Hwang, Chin-fa; Lin, Xin-yi; Yang, Fan-chiang; |
期刊 | Journal of the Chinese Institute of Chemical Engineers |
出版日期 | 20051100 |
卷期 | 36:6 民94.11 |
頁次 | 頁617-626 |
分類號 | 368.87 |
語文 | eng |
關鍵詞 | 酵母菌; γ-麩氨半胱氨酸; 抗氧化; 麩胱甘肽; γ-glutamylcysteine; γ-GC; Glutathione; GSH; Saccharomyces cerevisiae; Methylglyoxal; |
中文摘要 | γ-麩氨半胱氨酸(γ-GC)是人體重要抗氧化成分麩胱甘?(GSH)之前驅物質。本研究之目的為進行酵母菌之突變,藉以提高變異株中γ-GC之含量。首先歸選對甲基乙二醛(MG)敏感之變異株,接著分析弈異株之酵素活性及培養基組成對菌體及γ-GC生產之影響。結果顯示,在對MG敏感之變異株中篩選到一株黨號SY-35679菌株,其胞內γ-GC之含量提高到每克乾細胞中含4.77毫克,約為原始菌株(每克乾細胞中含0.59毫克)之八倍,而胞內GSH結量沒有明顯變化。比較原始菌株與突變株之生合成過程中重要酵素GSHⅠ及GSHⅡ活性之變化,發現突變株SY-35679之GSHⅠ酵素比活性明顯增高,而GSHⅡ之比活性則則沒有明顯差別。除外,葡萄糖之濃度與前瞻物質半胱氨酸(L-Cys)之添加影響菌體生長及γ-GC之產能。每升20克之葡萄糖明顯促進GSH及γ-GC之生產,而適當時機添加適當濃度之L-Cys對胞內含硫氫物質之增加具有正面之效果。 |
英文摘要 | γ-glutamylcysteine (γ-GC) is an essential precursor for the biosynthesis of glutathione (GSH). The formation ofγ-GC has been considered as the rate-limiting intermediate. The purpose of this study was to enhance theγ-GC production by a mutant strain derived from Saccharomyces cerevisiae BCRC 21727. The mutant strains were screened by the strategy including selection of methylglyoxal (MG) sensitive mutants through ethyl methanesulfonate (EMS) mutation and population analysis. The mutant’s characteristics and the effects of media compositions on the cell growth andγ-GC production were then studied. The results showed that a significantly highγ-GC-producing mutant, SY-35679, was isolated from the MG-sensitive mutants. The intracellularγ-GC content in the mutant SY-335679 rose to 4.77 mg.□ DCR and had around eight folds increase, compared with that of the parent strain (0.59 mg□DCW), while GSH content was at a similar level inside the mutant cell. The biosynthetic enzymes, GSHⅠ and GSHⅡ ,were compared between the mutant and parent strain. The specific activity of GSHⅠ in the mutant SY-35679 was found to be significantly higher than that of the parent strain, however, the GSHⅡ specific activity maintained at a similar level. In additions, the intracellular γ-GC and GSH contents were highly affected by different concentrations of glucose and L-cysteine supplement. The medium containing glucose of 20 g. □ caused the significant increase in boy γ-GC and GSH production. The proper addition of L-cysteine was also porved to have a positive effect on the levels of γ-GC and GSH. |
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