利用响应面法优化毕赤酵母 SO 菌株生产耐热 T1 脂肪酶的物理条件。
Optimization of physical conditions for the production of thermostable T1 lipase in Pichia guilliermondii strain SO using response surface methodology.
机构信息
Enzyme and Microbial Technology Research Center, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
出版信息
BMC Biotechnol. 2017 Nov 10;17(1):78. doi: 10.1186/s12896-017-0397-7.
BACKGROUND
Pichia guilliermondii was found capable of expressing the recombinant thermostable lipase without methanol under the control of methanol dependent alcohol oxidase 1 promoter (AOXp 1). In this study, statistical approaches were employed for the screening and optimisation of physical conditions for T1 lipase production in P. guilliermondii.
RESULT
The screening of six physical conditions by Plackett-Burman Design has identified pH, inoculum size and incubation time as exerting significant effects on lipase production. These three conditions were further optimised using, Box-Behnken Design of Response Surface Methodology, which predicted an optimum medium comprising pH 6, 24 h incubation time and 2% inoculum size. T1 lipase activity of 2.0 U/mL was produced with a biomass of OD 23.0.
CONCLUSION
The process of using RSM for optimisation yielded a 3-fold increase of T1 lipase over medium before optimisation. Therefore, this result has proven that T1 lipase can be produced at a higher yield in P. guilliermondii.
背景
毕赤酵母被发现能够在甲醇依赖型醇氧化酶 1 启动子（AOXp1）的控制下表达重组耐热脂肪酶，而无需甲醇。在这项研究中，采用统计方法筛选和优化毕赤酵母中 T1 脂肪酶生产的物理条件。
结果
通过 Plackett-Burman 设计对六种物理条件进行筛选，确定 pH 值、接种量和培养时间对脂肪酶生产有显著影响。利用响应面法的 Box-Behnken 设计对这三个条件进行了进一步优化，预测出最佳培养基组成包括 pH 值 6、24 小时培养时间和 2%接种量。在 OD23.0 的生物量下，T1 脂肪酶活性达到 2.0 U/mL。
结论
使用 RSM 进行优化的过程使 T1 脂肪酶的产量比优化前的培养基提高了 3 倍。因此，这一结果证明了 T1 脂肪酶可以在毕赤酵母中更高产。
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