利用 DTT（二硫苏糖醇）处理的芽孢杆菌 APB-6 休眠细胞合成乙酰胺肟酸的改进生物工艺。
An improved bioprocess for synthesis of acetohydroxamic acid using DTT (dithiothreitol) treated resting cells of Bacillus sp. APB-6.
机构信息
Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla 171005, India.
出版信息
Bioresour Technol. 2011 Jun;102(11):6579-86. doi: 10.1016/j.biortech.2011.03.071. Epub 2011 Apr 12.
Acyltransferase activity of amidase from Bacillus sp. APB-6 was enhanced (24 U) by multiple feedings of N-methylacetamide (70 mM) into the production medium. Hyperinduced whole resting cells of Bacillus sp. APB-6 corresponding to 4 g/L (dry cell weight), when treated with 10mM DTT (dithiothreitol) resulted in 93% molar conversion of acetamide (300 mM) to acetohydroxamic acid in presence of hydroxylamine-HCl (800 mM) after 30 min at 45°C in a 1L reaction mixture. After lyophilization, a 62 g powder containing 34% (wtwt(-1)) acetohydroxamic acid was recovered. This is the first report where DTT has been used to enhance acyltransfer reaction and such high molar conversion (%) of amide to hydroxamates was recorded at 1L scale.
来自芽孢杆菌 APB-6 的酰胺酶的酰基转移酶活性通过多次添加 N-甲基乙酰胺（70mM）到生产培养基中得到增强（24U）。当用 10mM DTT（二硫苏糖醇）处理时，超诱导的芽孢杆菌 APB-6 的全休眠细胞对应于 4g/L（干细胞重量），导致在 45°C 下在 1L 反应混合物中在 30 分钟内将乙酰胺（300mM）以 93%摩尔转化率转化为羟胺盐酸盐（800mM）。经冷冻干燥后，回收得到 62g 粉末，其中含有 34%（wt/wt(-1)）的羟胺盐酸盐。这是首次报道 DTT 用于增强酰基转移反应，并且在 1L 规模下记录到酰胺到羟胺酸盐的如此高的摩尔转化率（%）。
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