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. 2013 Sep 10;6:128. doi: 10.1186/1754-6834-6-128

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Copyright © 2013 Mattam and Yazdani; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Expression of clostridial pathway enzymes in E. coli for conversion of butyric acid to butanol. (A) The DH5α strain containing test and control plasmids were grown in LB medium in presence or absence of IPTG and analyzed for the expression of aldehyde/alcohol dehydrogenase (AdhE2) and butyrate kinase (Buk) containing 6-histidine tag on Western blot. Lane M – Molecular weight marker; lane 1 – pQE30 – IPTG; lane 2 - pQE30 + IPTG; lane 3 – pQE-adhE2/ptb/buk – IPTG; lane 4 – pQE-adhE2/ptb/buk + IPTG. (B) The grown cells in the LB medium were permeabilized with chloroform and analyzed for the activity of phosphotransbutyrylase (Ptb), Buk and AdhE2. (C) Cells containing control and test plasmids were grown in LB medium containing 10 mM butyric acid and samples were withdrawn after 48 h and 120 h to test for butanol production.