期刊
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
卷 106, 期 2, 页码 199-203出版社
SOC BIOSCIENCE BIOENGINEERING JAPAN
DOI: 10.1263/jbb.106.199
关键词
Rhodococcus opacus B-4; organic-solvent tolerant bacteria; alkB genes; anhydrous organic solvents; n-alkanes
资金
- New Energy and Industrial Technology Development Organization (NEDO
- Kawasaki)
- Research Project in the field of Biotechnology under the Ministry of Education, Culture, Sports, Science and Technology, Japan
- National Research Council of Thailand
- National Science and Technology Development Agency of Thailand
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 JM 109 showed that they encode functional alkane hydroxylases with a substrate range of C-5-C-16. Promoters of the alkB1 and alkB2 genes, designated P-alkB1 and P-alkB2, respectively, were examined for activity in anhydrous his (2-ethylhexyl) phthalate (BEHP) containing C-5-C-16 n-alkanes. Two recombinant plasmids, pP(alkB1)EGFP and pP(alkB2)EGFP, were constructed by inserting the egfp gene downstream of P-alkB1 and P-alkB2, respectively and transformed into R. opacus B-4. Resting cells of R. opacus B-4 (pP(alkB1)EGFP) showed greater levels of EGFP fluorescence in anhydrous BEHP than in 0.85% NaCl, when C-8-C-16 n-alkanes were supplied as an inducer. Furthermore, n-alkane inducibility of P-alkB1 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.
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