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Enhancing n-Butanol Tolerance of Escherichia coli by Overexpressing of Stress-Responsive Molecular Chaperones

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Abstract

Microbial tolerance to organic solvents is critical for efficient production of biofuels. In this study, n-butanol tolerance of Escherichia coli JM109 was improved by overexpressing of genes encoding stress-responsive small RNA-regulator, RNA chaperone, and molecular chaperone. Gene rpoS, coding for sigma S subunit of RNA polymerase, was the most efficient in improving n-butanol tolerance of E. coli. The highest OD600 and the specific growth rate of JM109/pQE80L-rpoS reached 1.692 and 0.144 h–1 respectively at 1.0% (v/v) n-butanol. Double and triple expression of molecular chaperones rpoS, secB, and groS were conducted and optimized. Recombinant strains JM109/pQE80L-secB-rpoS and JM109/pQE80L-groS-secB-rpoS exhibited the highest n-butanol tolerance, with specific growth rates of 0.164 and 0.165 h–1, respectively. Membrane integrity, potentials, and cell morphology analyses demonstrated the high viability of JM109/pQE80L-groS-secB-rpoS. This study provides guidance on employing various molecular chaperones for enhancing the tolerance of E. coli against n-butanol.

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Acknowledgments

We are grateful to the National Key Research and Development Program (2018YFA0901700), the National Natural Science Foundation of China (21776112), Natural Science Foundation of Jiangsu Province (BK20171135), the national first-class discipline program of Light Industry Technology and Engineering (LITE2018-07), the Program of Introducing Talents of Discipline to Universities (111-2-06), and Top-notch Academic Programs Project of Jiangsu Higher Education Institutions for the financial support of this research.

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  1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, 214122, China

    Guochao Xu, Lin Xiao, Anning Wu, Ruizhi Han & Ye Ni

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  1. Guochao Xu
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Xu, G., Xiao, L., Wu, A. et al. Enhancing n-Butanol Tolerance of Escherichia coli by Overexpressing of Stress-Responsive Molecular Chaperones. Appl Biochem Biotechnol 193, 257–270 (2021). https://doi.org/10.1007/s12010-020-03417-4

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