Bacillussp.肌氨酸氧化酶的脱辅基与重构

Deflavination ofBacillussp. Sarcosine Oxidase and Reconstitution of Flavoproteins

DOI：10.3969/j.issn.1673-1689.2018.09.003

中文关键词: 肌氨酸氧化酶 脱辅酶 透析复性 复性条件 重构

英文关键词: sarcosine oxidase,deflavination,dialysis,refolding condition,reconstitution

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中文摘要:

在肌氨酸氧化酶脱辅基的效果不够理想的情况下，利用不含辅基的重组肌氨酸氧化酶包涵体进行透析复性，以期重现SOX酶与辅酶的复合。研究表明，肌氨酸氧化酶（SOX）包涵体较适合的复性条件为：酶蛋白质量浓度0.3 g/L，透析液pH 8.5，温度4 ℃，氧化还原值（GSH/GSSG）为2，复性时间为24 h。作者还比较了不同天然辅基与SOX蛋白的复合效果，通过圆二色性、红外光谱以及DSC热差分析，探究了天然辅基与肌氨酸氧化酶复合后对酶蛋白二级结构、比酶活及稳定性的影响。

英文摘要:

Due to the unsatisfactory effects of deflavination from sarcosine oxidase（SOX），the SOX inclusion protein without cofactor was dialysed and renatured for testing the reconstruction effects between SOX apoprotein and the coenzymes. The comparatively suitable conditions for the binding of flavoprotein to SOX apoprotein were shown as following：the concentration of enzyme protein 0.3 g/L， pH 8.5， temperature 4 ℃， the optimal redox state（GSH / GSSG ） GSH / GSSG with the value 2，and the performing time was 24 h. The reconstitution effects of SOX protein binding to different natural flavins were compared. Through CD，FTIR，and DSC analysis，the secondary structure，enzyme specific activity and stability of the reconstituted SOX were also studied.

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