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Expression of Rhodococcus opacus alkB genes in anhydrous organic solvents

Paper ID Volume ID Publish Year Pages File Format Full-Text
21661 43233 2008 5 PDF Available
Title
Expression of Rhodococcus opacus alkB genes in anhydrous organic solvents
Abstract

Rhodococcus opacus B-4 is a benzene-tolerant bacterium which was isolated from a gasoline-contaminated soil sample. We previously demonstrated that this organism was able to survive and exhibit biocatalytic activity in anhydrous organic solvents for at least 5 d. In the present study, we cloned the alkB1 and alkB2 genes encoding alkane hydroxylases from R. opacus B-4. Heterologous expression of the alkB1 and alkB2 genes in Escherichia coli JM109 showed that they encode functional alkane hydroxylases with a substrate range of C5–C16. Promoters of the alkB1 and alkB2 genes, designated PalkB1 and PalkB2, respectively, were examined for activity in anhydrous bis (2-ethylhexyl) phthalate (BEHP) containing C5–C16n-alkanes. Two recombinant plasmids, pPalkB1EGFP and pPalkB2EGFP, were constructed by inserting the egfp gene downstream of PalkB1 and PalkB2, respectively and transformed into R. opacus B-4. Resting cells of R. opacus B-4 (pPalkB1EGFP) showed greater levels of EGFP fluorescence in anhydrous BEHP than in 0.85% NaCl, when C8–C16n-alkanes were supplied as an inducer. Furthermore, n-alkane inducibility of PalkB1 activity in anhydrous BEHP was noticeably different from that in 0.85% NaCl. This paper presents the first evidence that bacteria can express their genes in essentially anhydrous organic solvents.

Keywords
Rhodococcus opacus B-4; organic-solvent tolerant bacteria; alkB genes; anhydrous organic solvents; n-alkanes
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Expression of Rhodococcus opacus alkB genes in anhydrous organic solvents
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 106, Issue 2, August 2008, Pages 199–203
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us