Solvent resistance pumps of Pseudomonas putida S12: Applications in 1-naphthol production and biocatalyst engineering.
In: Journal of Biotechnology, Jg. 210 (2015-09-20), S. 91-99
academicJournal
Zugriff:
The solvent resistance capacity of Pseudomonas putida S12 was applied by using the organism as a host for biocatalysis and through cloning and expressing solvent resistant pump genes into Escherichia coli . P. putida S12 expressing toluene ortho mononooxygenase (TOM-Green) was used for 1-naphthol production in a water-organic solvent biphasic system. Application of P. putida S12 improved 1-naphthol production per gram cell dry weight by approximately 42% compared to E. coli . Moreover, P. putida S12 enabled the use of a less expensive solvent, decanol, for 1-naphthol production. The solvent resistant pump (srpABC) genes of P. putida S12 were cloned into a solvent sensitive E. coli strain to transfer solvent tolerance. Recombinant strains bearing srpABC genes in either a low-copy number or a high-copy number plasmid grew in the presence of saturated concentration of toluene. Both of the recombinant strains were more tolerant to 1% v/v of toxic solvents, decanol and hexane, reaching similar cell density as the no-solvent control. Reverse-transcriptase analysis revealed that the srpABC genes were transcribed in engineered strains. The results demonstrate successful transfer of the proton-dependent solvent resistance mechanism and suggest that the engineered strain could serve as more robust biocatalysts in media with organic solvents. [ABSTRACT FROM AUTHOR]
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Solvent resistance pumps of Pseudomonas putida S12: Applications in 1-naphthol production and biocatalyst engineering.
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Autor/in / Beteiligte Person: | Janardhan Garikipati, S.V.B. ; Peeples, Tonya L. |
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Zeitschrift: | Journal of Biotechnology, Jg. 210 (2015-09-20), S. 91-99 |
Veröffentlichung: | 2015 |
Medientyp: | academicJournal |
ISSN: | 0168-1656 (print) |
DOI: | 10.1016/j.jbiotec.2015.06.419 |
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