响应面法优化热带假丝酵母104菌株产羰基还原酶发酵培养基_王普.pdfVIP

生 物 工 程 学 报 Chin J Biotech 2009, June 25; 25(6): 863-868 journals.im.ac.cn Chinese Journal of Biotechnology ISSN 1000-3061 cjb@im.ac.cn © 2009 Institute of Microbiology, CAS CSM, All rights reserved 工业生物技术 104 王普, 孙立明, 何军邀 浙江工业大学药学院, 杭州 310032 摘 要: 通过菌种优选得到产高选择性羰基还原酶的热带假丝酵母(Candida tropicalis) 104 菌株, 可不对称还原 1-(3,5- 二三氟甲基)苯基乙酮生成(S)-1-(3,5-二三氟甲基)苯基乙醇, 对映体过量值99.9% 。采用部分因子实验设计考察发酵培 养基中各组分对产酶的影响, 结果表明, 酵母粉、葡萄糖和NH4Cl 浓度对产酶影响显著。继而采用最陡爬坡路径逼近最 大响应区域, 并结合中心组合实验和响应面对3 个显著性因素进行分析, 得到优化的发酵培养基组成: 葡萄糖47.14 g/L, 酵母粉 13.25 g/L, NH Cl 2.71 g/L, MgSO ·7H O 0.4 g/L, KH PO 1 g/L 和K HPO 1 g/L 。采用该优化培养基, 供试菌株的 4 4 2 2 4 2 4 羰基还原酶活力达 851.13 U/L, 较优化前提高了 65.2% 。 关键词: 热带假丝酵母, 羰基还原酶, (S)-1-(3,5-二三氟甲基)苯基乙醇, 响应面, 培养基优化 Medium optimization for enhanced production of carbonyl reductase by Candida tropicalis 104 by response surface methodology Pu Wang, Liming Sun, and Junyao He College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310032, China Abstract: Using response surface method, we optimized the medium for the asymmetric whole cell biotransformation by Candida tropicalis 104. This strain was used for microbial reduction of 1-[3,5-bis(trifluoromethyl)phenyl] ethanone to (S)-1-[3,5-bis(trifluoromethyl)phenyl] alcohol, with enantiomeric excess(e.e.) reached more than 99.9%. Fractional factorial design was used to evaluate the effects of medium compo