Auto-induction-based Rapid Evaluation of Extracellular Enzyme Expression from Lac Operator-involved Recombinant Escherichia coli
Auto-induction-based Rapid Evaluation of Extracellular Enzyme Expression from Lac Operator-involved Recombinant Escherichia coli
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基于自诱导的 Lac 操纵子重组大肠杆菌胞外酶表达的快速评估
DOI：
10.1007/s12010-014-1201-y
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发表时间：
2014-09
影响因子：
3
通讯作者：
Rong Xiao
中科院分区：
文献类型：
--
作者：
Wen-Bo Chen;Yao Nie;Xiao-Qing Mu;Wei Yan;Yan Xu;Rong Xiao
在重组生物过程中，经常观察到工程菌株的退化和目标基因产物的产量下降。虽然已经开发了几种策略来确保高水平的靶基因产物，但这些方法似乎是复杂和费力的。通过研究导致产量下降的可能因素，确定了宿主细胞的退化是主要原因。基于自诱导原理和有色底物与基因产物相互作用可产生明显变化的特点，开发了一种简便、准确的筛选方法来评价工程菌株的生产性能。由于克雷伯氏菌普鲁兰酶的典型特征，它已成为研究革兰氏阴性菌分泌机制的模型，本文采用一种产生胞外普鲁兰酶的工程大肠杆菌来说明该方法。因此，根据形成无色光晕的能力，可以将菌落分为两类，一类被透明带包围，另一类不能呈现透明光晕。此外，通过在液体培养基中进行普鲁兰酶生产，证实了这些菌落的高能力。与其他评价工程菌株的方法相比，可见筛选技术具有有效和准确的优点。
Degeneration of engineered strains and decreased production of target gene products were often observed during recombinant bioprocess. Although several strategies have been developed to ensure high levels of target gene products, these methods seemed to be complex and laborious. By investigating possible factors contributing to the decreased yield, degeneration of host cells was identified as the main cause. Based on the principle of auto-induction and the fact that the interaction between colored substrates and gene products could present visible changes, a convenient and accurate screening method was developed to evaluate the production performance of engineered strains. Due to the typicality of pullulanase from genus Klebsiella, which has been a model for the research of secretion mechanism in gram-negative bacteria, an engineered E. coli producing extracellular pullulanase was employed to illustrate the method. Consequently, according to the capability to form a colorless halo, colonies could be divided into two groups, one surrounded by clear zone and the other unable to present transparent haloes. Furthermore, the high capability of these colonies was confirmed by performing pullulanase production in liquid medium. Compared with other methods for evaluating engineered strains, the visible screening technique was suggested to have the advantages of effectiveness and accuracy.
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影响因子：
6.4
作者：
Su L;Chen S;Yi L;Woodard RW;Chen J;Wu J
通讯作者：
Wu J
影响因子：
5
作者：
Fu, Xiang-Yang
通讯作者：
Fu, Xiang-Yang
影响因子：
4.1
作者：
B. Sommer;K. Friehs;E. Flaschel
通讯作者：
B. Sommer;K. Friehs;E. Flaschel
影响因子：
5.6
作者：
Hii, Ling S.;Rosfarizan, Mohamad;Ariff, Arbakariya B.
通讯作者：
Ariff, Arbakariya B.
影响因子：
3.8
作者：
Chen, Wen-Bo;Nie, Yao;Xiao, Rong
通讯作者：
Xiao, Rong