Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China.
Enzyme Microb Technol. 2011 Sep 10;49(4):321-5. doi: 10.1016/j.enzmictec.2011.06.008. Epub 2011 Jun 15.
This work aims to produce α-ketoisocaproate (KIC) from L-leucine via the free-whole-cell biotransformation of Rhodococcus opacus DSM 43250. The effects of temperature, pH, substrate concentration, cell concentration, and rotating speed on KIC production were examined. Furthermore, the biotransformation conditions were optimized with response surface methodology (RSM). The optimal biotransformation conditions were as follows: temperature 43.7 °C, pH 8.4, L-leucine concentration 5.1g/L, cell concentration 30.4 g/L, and rotating speed 170 rpm. The maximal KIC production predicted by RSM model reached 1275 mg/L at the optimal conditions, and in the validated experiments, the maximal KIC production reached 1264 mg/L, which was very close to the predicted KIC production. The current work provides an alternative for the production of KIC and the results obtained here are useful for the biotransformatic KIC production on a large scale.
本工作旨在通过罗特氏球红球菌 DSM 43250 的全细胞自由生物转化从 L-亮氨酸生产 α-酮异己酸(KIC)。考察了温度、pH 值、底物浓度、细胞浓度和转速对 KIC 生产的影响。此外,还采用响应面法(RSM)对生物转化条件进行了优化。最佳生物转化条件为:温度 43.7°C,pH 值 8.4,L-亮氨酸浓度 5.1g/L,细胞浓度 30.4 g/L,转速 170 rpm。RSM 模型预测的最佳条件下 KIC 的最大产量达到 1275mg/L,在验证实验中,KIC 的最大产量达到 1264mg/L,非常接近预测的 KIC 产量。本工作为 KIC 的生产提供了一种替代方法,所得结果对大规模生物转化生产 KIC 具有实用价值。