Zhao Jie, Wang Yonghong, Chu Ju, Zhang Siliang, Zhuang Yingping, Yuan Zhongyi
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237 Shanghai, People's Republic of China.
J Ind Microbiol Biotechnol. 2008 Apr;35(4):257-62. doi: 10.1007/s10295-007-0301-x. Epub 2008 Jan 31.
Pyruvate oxidase (PyOD) is a very useful enzyme for clinical diagnostic applications and environmental monitor. Optimization of the fermentation medium for maximization of PyOD constitutively, production by Escherichia coli DH5alpha/pSMLPyOD was carried out. Response surface methodology (RSM) was used to optimize the medium constituents. A 2(6-2) fractional factorial design (first order model) was carried out to identify the significant effect of medium components towards PyOD production. Statistical analysis of results shows that yeast extract, ammonium sulfate and composite phosphate were significant factors on PyOD production. The optimized values of these three factors were obtained by RSM based on the result of a 2(3) central composite rotatable design. Under these proposed optimized medium, the model predicted a PyOD activity of 610 U/L and via experimental rechecking the model, an activity of 670 U/L was attained.
丙酮酸氧化酶（PyOD）是一种在临床诊断应用和环境监测中非常有用的酶。通过大肠杆菌DH5alpha/pSMLPyOD组成型生产PyOD以实现发酵培养基的优化。采用响应面法（RSM）优化培养基成分。进行了2（6-2）部分因子设计（一阶模型）以确定培养基成分对PyOD生产的显著影响。结果的统计分析表明，酵母提取物、硫酸铵和复合磷酸盐是影响PyOD生产的显著因素。基于2（3）中心复合旋转设计的结果，通过RSM获得了这三个因素的优化值。在这些建议的优化培养基条件下，模型预测的PyOD活性为610 U/L，通过对模型的实验重新验证，获得了670 U/L的活性。
