Abstract
A 3-hydroxyisobutyrate dehydrogenase-encoding gene mmsB has been identified as one of the key genes responsible for the enhanced organic solvent tolerance (OST) of Pseudomonas putida JUCT1. In this study, the OST-related effect of two 3-hydroxyacid dehydrogenase family genes (mmsB and zwf) was investigated in Escherichia coli JM109. It was noted that the growth of E. coli JM109 was severely hampered in 4 % decalin after zwf knockout. Additionally, its complementation resulted in significantly enhanced solvent tolerance compared with its parent strain. Furthermore, E. coli JM109 carrying mmsB showed better OST capacity than that harboring zwf. To construct E. coli strains with an inheritable OST phenotype, mmsB was integrated into the genome of E. coli JM109 by red-mediated recombination. Using E. coli JM109(DE3) (ΔendA::mmsB) as host strain, whole-cell biocatalysis was successfully carried out in an aqueous/butyl acetate biphasic system with a remarkably improved product yield.
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Acknowledgments
We are grateful to the Natural Science Foundation of China (21276112), National Basic Research and Development Program of China (973 Program, 2011CB710800), New Century Excellent Talents in University (NCET-11-0658), Natural Science Foundation of Jiangsu Province (BK2011150), the Program of Introducing Talents of Discipline to Universities (111-2-06), and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions for the financial support of this research.
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Qian, X., Song, L. & Ni, Y. Enhanced Organic Solvent Tolerance of Escherichia coli by 3-Hydroxyacid Dehydrogenase Family Genes. Appl Biochem Biotechnol 172, 3106–3115 (2014). https://doi.org/10.1007/s12010-014-0726-4
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DOI: https://doi.org/10.1007/s12010-014-0726-4