Abstract
A portion of the cbpA gene from Escherichia coli K-12 encoding a 24 amino acid proton-buffering peptide (Pbp) was cloned via the shuttle vector pJB99 into E. coli JM105 and subsequently into Zymomonas mobilis CP4. Expression of Pbp was confirmed in both JM105 and CP4 by HPLC. Z. mobilis CP4 carrying pJB99-2 (Pbp) exhibited increased acid tolerance (p<0.05) in acidified TSB (HCl [pH 3.0] or acetic acid [pH 3.5]), glycine-HCl buffer (pH 3.0), and sodium acetate-acetic acid buffer (pH 3.5) in comparison to the parent strain (CP4) and CP4 with pJB99 (control plasmid). Although the expression of Pbp influenced survival at a low pH, the minimum growth pH was unaffected. Growth of Z. mobilis in the presence of ampicillin also significantly increased acid tolerance by an unknown mechanism. Results from this study demonstrate that the production of a peptide with a high proportion of basic amino acids can contribute to protection from low pH and weak organic acids such as acetic acid.
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Dien, B. S., Cotta, M. A., and Jeffries, T. W. (2003) Appl. Microbiol. Biotechnol. 63, 258–266.
Jeffries, T. W. and Jin, Y. S. (2004) Appl. Microbiol. Biotechnol. 63, 495–509.
Picataggio, S. K., Zhang, M., and Finkelstein, M. (1994), in Enzymatic Conversion of Biomass For Fuels Production (Himmel, M. E., Baker, J. O., and Overend, R. P., eds.), ACS Symp. Ser. 566, 342–362.
Swings, J. and DeLey, J. (1977) Bacteriol. Rev. 41, 1–46
Stokes, H. W., Picataggio, S. K., and Eveleigh, D. E. (1983), in Advances in Solar Energy (Böer, K. W. and Duffie, J. A., eds.), American Solar Energy Society, New York, NY, pp. 113–132.
Zhang, M., Eddy, C., Deanda, K., Finkelstein, M., and Picataggio, S. (1995) Science. 267, 240–243.
Mohagheghi, A., Evans, K., Chou, Y. C., and Zhang, M. (2002) Appl. Biochem. Biotechnol. 98–100, 885–898.
Joachimsthal, E. L., Haggett, K. D., and Rogers, P. L. (1999) Appl. Biochem. Biotechnol. 77, 147–157.
Kim, I. S., Barrow, K. D., and Rogers, P. L. (2000) Appl. Biochem. Biotechnol. 84–86, 357–370.
Lawford, H. G. and Rousseau, J. D. (1999) Appl. Biochem. Biotechnol. 77–79, 235–249.
Tucker, D. L., Tucker, N., and Conway, T. (2002) J. Bacteriol. 184, 6551–6558.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: a Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Arnold, K. W. and Kaspar, C. W. (1995) Appl. Environ. Microbiol. 61, 2037–2039.
Byrd, J. J., Cheville, A. M., Bose, J. L., and Kaspar, C. W. (1999) Appl. Environ. Microbiol. 65, 2396–2401.
Afendra, A. S. and Drainas, C. (1987). J. Gen. Microbiol. 133, 127–134.
Seo, J. S., Chong, H., Park, H. S., Yoon, K. O., Jung, C., Kim, J. J., Hong, J. H., et al. (2005) Nature Biotech. 23(1) 63–68.
Lehninger, A. L., Nelson, D. L., Cox, M. M. (1993), in Principles of Biochemistry, 2nd ed., Worth Publishers, New York, NY, p. 113.
Creighton, T. E. (1993) Proteins: Structures and Molecular Properties, 2nd ed., W. H. Freeman and Co., New York, NY.
Belaich, J. P. and Senez, J. C. (1965) J. Bacteriol. 89, 1195–1200.
Foster, J. W. (2000), in Bacterial Stress Responses (Storz, G., and Hengge-Aronis, R., eds.), American Society for Microbiology Press, Washington, D.C., pp. 99–115.
Sutcliffe, J. G. (1978) Proc. Natl. Acad. Sci. USA 75(8), 3737–3741.
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Baumler, D.J., Hung, K.F., Bose, J.L. et al. Enhancement of acid tolerance in Zymomonas mobilis by a proton-buffering peptide. Appl Biochem Biotechnol 134, 15–26 (2006). https://doi.org/10.1385/ABAB:134:1:15
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DOI: https://doi.org/10.1385/ABAB:134:1:15