Hee Jin Hwang and Jin Hwan Park contributed equally to this work.
Engineering of a butyraldehyde dehydrogenase of Clostridium saccharoperbutylacetonicum to fit an engineered 1,4-butanediol pathway in Escherichia coli
Article first published online: 24 FEB 2014
© 2014 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 111, Issue 7, pages 1374–1384, July 2014
How to Cite
Hwang, H. J., Park, J. H., Kim, J. H., Kong, M. K., Kim, J. W., Park, J. W., Cho, K. M. and Lee, P. C. (2014), Engineering of a butyraldehyde dehydrogenase of Clostridium saccharoperbutylacetonicum to fit an engineered 1,4-butanediol pathway in Escherichia coli. Biotechnol. Bioeng., 111: 1374–1384. doi: 10.1002/bit.25196
- Issue published online: 29 MAY 2014
- Article first published online: 24 FEB 2014
- Accepted manuscript online: 22 JAN 2014 05:06AM EST
- Manuscript Accepted: 17 JAN 2014
- Manuscript Revised: 12 JAN 2014
- Manuscript Received: 3 OCT 2013
- Samsung Advanced Institute of Technology
- National Research Foundation of Korea
- Korean Government. Grant Numbers: 2011-0030336, 2012M1A2A2026562
Additional supporting information may be found in the online version of this article at the publisher's web-site.
Figure S1. Optical density at 540 m values of five selected butyraldehyde dehydrogenase (Bld) mutants mixed with Schiff's reagent.
Figure S2. Sequence alignment between Bld and two template proteins (aldehyde dehydrogenase of Listeria monocytogenes [Protein Data Bank ID: 3K9D] and alcohol dehydrogenase of Vibrio parahaemolyticus [Protein Data Bank ID: 3MY7]). Sequence alignment of Bld to 3K9D and 3MY7 sequences was performed for homology modeling. The results showed 15.0% sequence identity and 34.1% sequence similarity. Conserved catalytic cysteine residues (Cys275 in the case of Bld) are indicated with a red asterisk.
Table SI. Primers used for genome engineering.
Table SII. Cloning polymerase chain reaction primers for 1,4-butanediol pathway enzymes.
Table SIII. Primers used for site-directed mutagenesis.
Table SIV. Butyraldehyde dehydrogenase (Bld) L273X generated through site-directed mutagenesis.
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