| 乙醇发酵中酿酒酵母辅酶NAD+及NADH测定方法
| Determination of Coenzyme NAD~+ and NADH of Saccharomyces cerevisiae Cells in Ethanol Production
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| DOI：
| 中文关键词: 酿酒酵母 NAD+/NADH 珠磨破碎法 酶循环法
| 英文关键词: Saccharomyces cerevisiae NAD+/NADH shaking bead enzymatic cycling
| 基金项目:上海市重点学科建设项目(B505)
| 作者
| 单位
| 李骆冰
| 华东理工大学,生物反应器工程国家重点实验室,上海,200237
| 王永红
| 华东理工大学,生物反应器工程国家重点实验室,上海,200237
| 庄英萍
| 华东理工大学,生物反应器工程国家重点实验室,上海,200237
| 储炬
| 华东理工大学,生物反应器工程国家重点实验室,上海,200237
| 张嗣良
| 华东理工大学,生物反应器工程国家重点实验室,上海,200237
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| 中文摘要:
| 乙醇发酵过程中酿酒酵母细胞辅酶NAD+和NADH质量分数及其比例反映了胞内的氧化还原状态和细胞代谢活性,具有重要的生理意义。作者主要研究了加热萃取、珠磨破碎、反复冻融破碎3种方法对乙醇发酵过程酿酒酵母细胞辅酶NAD+和NADH提取和测定的影响,提出基于珠磨破碎、辅以加酸或碱并加热的萃取模式,以及合适的酶循环反应体系,从而建立了高效的胞内NAD+和NADH检测方法。
| 英文摘要:
| Due to the importance of NAD+ and NADH levels in the yeast metabolism,the determination of intracellular NADH/NAD+ is a very interesting for ethanol production.In this study,Three NADH./NAD+ extracting methods,including heating,freeze-thaw cycles,shaking bead,for the disruption and extraction of the NAD+ and NADH in yeast cells were investigated and compared.Finally an efficient process for extraction and detection of NAD+ and NADH in yeast cells was established as follows.Yeast cells were disrupted by shaking bead while NAD+ and NADH were extracted by acid or alkali,then the extracts were detected under the favorable enzymatic cycling system Ⅱ.
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