Optimization of cell culture and cell disruption processes to enhance the production of thermophilic cellulase FnCel5A in E.coli using response surface methodology
Optimization of cell culture and cell disruption processes to enhance the production of thermophilic cellulase FnCel5A in E.coli using response surface methodology
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使用响应面方法优化细胞培养和细胞破碎过程，以提高大肠杆菌中嗜热纤维素酶 FnCel5A 的产量
DOI：
10.1371/journal.pone.0210595
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发表时间：
2019-01
期刊：
影响因子：
3.7
通讯作者：
Yang Guangyu
中科院分区：
文献类型：
--
作者：
Mohammad Shah Faisal;Feng Yan;Yang Guangyu
来自结节热袍菌（Fervidobacterium nodosum）的FnCel5A是最耐热的内切葡聚糖酶之一，具有显著的特性，例如高活性、pH稳定性以及对多种底物的多特异性。然而，大规模的嗜热酶产物……
FnCel5A from Fervidobacterium nodosum is one of the most thermostable endoglucanases that have phenomenal characteristics, such as high activity, pH stability, and multi-specificity towards various substrates. However, large-scale thermophilic enzyme production is still a challenge. Herein, we focus on an optimization approach based on response surface methodology to improve the production of this enzyme. First, a Box-Behnken design was used to examine physiochemical parameters such as induction temperatures, isopropylβ-D-1-thiogalactopyranoside concentrations and induction times on the heterogeneous expression of FnCel5A gene in E. coli. The best culture was collected after adding 0.56 mM IPTG and incubating it for 29.5 h at 24°C. The highest enzymatic activity observed was 3.31 IU/mL. Second, an economical "thermolysis" cell lysis method for the liberation of the enzymes was also optimized using Box-Behnken design. The optimal levels of the variables were temperature 77°C, pH 7.71, and incubation time of 20 min, which gave about 74.3% higher activity than the well-established bead-milling cell disruption method. The maximum productivity of FnCel5A achieved (5772 IU/L) illustrated that its production increased significantly after combining both optimal models. This strategy can be scaled-up readily for overproduction of FnCel5A from recombinant E.coli to facilitate its usage in biomass energy production.
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基于伴侣的程序，以增加大肠杆菌中产生的可溶性重组蛋白的产量。
影响因子：
3.5
作者：
通讯作者：
--
DOI：
--
发表时间：
2009
期刊：
--
影响因子：
--
作者：
R. Huber;D. Ritter;Till Hering;Anne-Kathrin Hillmer;F. Kensy;C. Müller;Le Wang;J. Büchs
通讯作者：
R. Huber;D. Ritter;Till Hering;Anne-Kathrin Hillmer;F. Kensy;C. Müller;Le Wang;J. Büchs
影响因子：
--
作者：
Yu Cao;De-jing Meng;Jian Lu;Jie Long
通讯作者：
Yu Cao;De-jing Meng;Jian Lu;Jie Long
影响因子：
4.4
作者：
OH, S;RHEEM, S;BAEK, Y
通讯作者：
BAEK, Y
影响因子：
4.1
作者：
Paes, Gabriel;O'Donohue, Michael J.
通讯作者：
O'Donohue, Michael J.