Bioprocess and Molecular Engineering Research Unit, Department of Biotechnology Engineering, Faculty of Engineering, International Islamic University Malaysia, P,O, Box 10, 50728, Kuala Lumpur, Malaysia.
AMB Express. 2012 Feb 15;2:12. doi: 10.1186/2191-0855-2-12.
Bromelain, a cysteine protease with various therapeutic and industrial applications, was expressed in Escherichia coli, BL21-AI clone, under different cultivation conditions (post-induction temperature, L-arabinose concentration and post-induction period). The optimized conditions by response surface methodology using face centered central composite design were 0.2% (w/v) L-arabinose, 8 hr and 25°C. The analysis of variance coupled with larger value of R2 (0.989) showed that the quadratic model used for the prediction was highly significant (p < 0.05). Under the optimized conditions, the model produced bromelain activity of 9.2 U/mg while validation experiments gave bromelain activity of 9.6 ± 0.02 U/mg at 0.15% (w/v) L-arabinose, 8 hr and 27°C. This study had innovatively developed cultivation conditions for better production of recombinant bromelain in shake flask culture.
菠萝蛋白酶是一种半胱氨酸蛋白酶,具有多种治疗和工业应用,在不同的培养条件(诱导后温度、L-阿拉伯糖浓度和诱导后时间)下在大肠杆菌 BL21-AI 克隆中表达。利用中心复合设计的响应面法对优化条件进行了优化,结果为 0.2%(w/v)L-阿拉伯糖、8 小时和 25°C。方差分析和较大的 R2 值(0.989)表明,用于预测的二次模型非常显著(p < 0.05)。在优化条件下,该模型产生的菠萝蛋白酶活性为 9.2 U/mg,而验证实验在 0.15%(w/v)L-阿拉伯糖、8 小时和 27°C 下得到的菠萝蛋白酶活性为 9.6 ± 0.02 U/mg。本研究创新性地开发了摇瓶培养中更好生产重组菠萝蛋白酶的培养条件。