Chou C P, Wang W C, Lin M I
Department of Chemical Engineering, Feng Chia University, Taichung, Taiwan, ROC.
Biotechnol Prog. 2000 May-Jun;16(3):315-8. doi: 10.1021/bp0000205.
Heterologous production of Providencia rettgeri penicillin acylase (PAC) was optimized in Escherichia coli. Several factors, including carbon, temperature, and host effects, were identified to be critical for the enzyme overproduction. The optimum culture conditions for the enzyme production vary for different host/vector systems. With the optimization, both volumetric and specific PAC activities could be significantly improved by more than 50-fold compared to the native expression in P. rettgeri. The heterologous production could be possibly limited by translation or posttranslational steps, depending on the culture temperature and host/vector system. To our knowledge, this is the first evidence demonstrating the limiting step for the production of P. rettgeri PAC and the existence of the P. rettgeri PAC precursor.
在大肠杆菌中对雷特格普罗威登斯菌青霉素酰化酶（PAC）的异源生产进行了优化。确定了包括碳源、温度和宿主效应在内的几个因素对该酶的过量生产至关重要。不同宿主/载体系统中酶生产的最佳培养条件各不相同。通过优化，与雷特格普罗威登斯菌中的天然表达相比，PAC的体积活性和比活性均可显著提高50倍以上。根据培养温度和宿主/载体系统的不同，异源生产可能受到翻译或翻译后步骤的限制。据我们所知，这是首个证明雷特格普罗威登斯菌PAC生产限制步骤以及雷特格普罗威登斯菌PAC前体存在的证据。
