Applied Microbiology and Biotechnology

, Volume 63, Issue 3, pp 293–299 | Cite as

Overexpression of the ATP-dependent helicase RecG improves resistance to weak organic acids in Escherichia coli

  • P. Steiner
  • U. SauerEmail author
Original Paper


Increased resistance to several weak organic acids was conferred on Escherichia coli by overexpression of the ATP-dependent helicase RecG and, to a lesser extent, by overexpressing the helicase RuvAB. This property of helicases was identified by reproducible selection of recG-bearing clones from genomic libraries of the acetate-resistant species Acetobacter aceti and Staphylococcus capitis. We show that overexpression of RecG from both species, but also from E. coli, increased the maximum biomass concentration attained by E. coli cultures that were grown in the presence of various weak organic acids and uncouplers. Furthermore, overexpression of RecG from A. aceti significantly improved the maximum growth rates of E. coli under weak organic acid challenge. Based on the known role of RecG in DNA replication/repair, our data provide a first indication that weak organic acids negatively affect DNA replication and/or repair, and that these negative effects may be counteracted by helicase activity.


Genomic Library Luria Broth Maximum Specific Growth Rate Restriction Enzyme Analysis Acetic Acid Bacterium 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank E. Marti for constructing the S. capitis library and for help with the selection experiment, H. Ernst for sequencing, and R. Stephan for providing chromosomal DNA of E. coli O157:H7. Funding from the ETH Forschungskommision is acknowledged.


  1. Aiba S, Humphrey AE, Millis NF (1973) Biochemical Engineering, 2nd edn. Academic Press, New YorkGoogle Scholar
  2. Audia JP, Webb CC, Foster JW (2001) Breaking through the acid barrier: an orchestrated response to proton stress by enteric bacteria. Int J Med Microbiol 291:97–106PubMedGoogle Scholar
  3. Brul S, Coote P (1999) Preservative agents in foods: mode of action and microbial resistance mechanisms. Int J Food Microbiol 50:1–17Google Scholar
  4. Cherrington CA, Hinton M, Mead GC, Chopra I (1991) Organic acids: chemistry, antibacterial activity and practical applications. Adv Microb Physiol 32:87–108PubMedGoogle Scholar
  5. Diez-Gonzalez F, Russell JB (1997) The ability of Escherichia coli O157:H7 to decrease its intracellular pH and resist the toxicity of acetic acid. Microbiology 143:1175–1180PubMedGoogle Scholar
  6. Dürre P, Bahl H, Gottschalk G (1988) Membrane processes and product formation in anaerobes. In: Erickson LE, Fung DYC (eds) Handbook for anaerobic fermentations. Dekker, New York, pp 187–206Google Scholar
  7. Emmerling M, Dauner M, Ponti A, Fiaux J, Hochuli M, Szyperski T, Wüthrich K, Bailey JE, Sauer U (2002) Metabolic flux responses to pyruvate kinase knockout in Escherichia coli. J Bacteriol 184:152–164CrossRefPubMedGoogle Scholar
  8. Foster JW (1995) Low pH adaptation and the acid tolerance response of Salmonella typhimurium. Crit Rev Microbiol 21:215–237PubMedGoogle Scholar
  9. Freese E, Sheu CW, Galliers E (1973) Function of lipophilic acids as antimicrobial food additives. Nature 241:321–325PubMedGoogle Scholar
  10. Hanna MN, Ferguson RJ, Li YH, Cvitkovitch DG (2001) uvrA is an acid-inducible gene involved in the adaptive response to low pH in Streptococcus mutans. J Bacteriol 183:5964–5973CrossRefPubMedGoogle Scholar
  11. Krebs HA, Wiggins D, Stubbs M, Sols A, Bedoya F (1983) Studies on the mechanism of the antifungal action of benzoate. Biochem J 214:657–663PubMedGoogle Scholar
  12. Kuroda M, Ohta T, Uchiyama I, Baba T, Yuzawa H, Kobayashi I, Cui L, Oguchi A, Aoki K, Nagai Y, Lian J, Ito T, Kanamori M, Matsumaru H, Maruyama A, Murakami H, Hosoyama A, Mizutani-Ui Y, Takahashi NK, Sawano T, Inoue R, Kaito C, Sekimizu K, Hirakawa H, Kuhara S, Goto S, Yabuzaki J, Kanehisa M, Yamashita A, Oshima K, Furuya K, Yoshino C, Shiba T, Hattori M, Ogasawara N, Hayashi H, Hiramatsu K (2001) Whole genome sequencing of meticillin-resistant Staphylococcus aureus. Lancet 357:1225–1240CrossRefPubMedGoogle Scholar
  13. Lasko DR, Schwerdel C, Bailey JE, Sauer U (1997) Acetate-specific stress response in acetate-resistant bacteria: an analysis of protein patterns. Biotechnol Prog 13:519–523CrossRefPubMedGoogle Scholar
  14. Lasko DR, Zamboni N, Sauer U (2000) The bacterial response to acetate challenge: a comparison of tolerance among species. Appl Microbiol Biotechnol 54:243–247PubMedGoogle Scholar
  15. Lee SJ, Gralla JD (2001) Sigma38 (rpoS) RNA polymerase promoter engagement via −10 region nucleotides. J Biol Chem 276:30064–30071CrossRefPubMedGoogle Scholar
  16. Lloyd RG, Sharples GJ (1991) Molecular organization and nucleotide sequence of the recG locus of Escherichia coli K-12. J Bacteriol 173:6837–6843PubMedGoogle Scholar
  17. Lloyd RG, Sharples GJ (1993) Processing of recombination intermediates by the RecG and RuvAB proteins of Escherichia coli. Nucleic Acids Res 21:1719–1725PubMedGoogle Scholar
  18. Mahdi AA, McGlynn P, Levett SD, Lloyd RG (1997) DNA binding and helicase domains of the Escherichia coli recombination protein RecG. Nucleic Acids Res 25:3875–3880CrossRefPubMedGoogle Scholar
  19. McGlynn P, Lloyd RG (2002) Genome stability and the processing of damaged replication forks by RecG. Trends Genet 18:413–419CrossRefPubMedGoogle Scholar
  20. Ochsner UA, Vasil ML, Alsabbagh E, Parvatiyar K, Hassett DJ (2000) Role of the Pseudomonas aeruginosa oxyR-recG operon in oxidative stress defense and DNA repair: oxyR-dependent regulation of katB-ankB, ahpB, and ahpC-ahpF. J Bacteriol 182:4533–4544CrossRefPubMedGoogle Scholar
  21. Park YS, Toda K, Fukaya M, Okumura H, Kawamura Y (1991) Production of a high-concentration acetic-acid by Acetobacter aceti using a repeated fed-batch culture with cell recycling. Appl Microbiol Biotechnol 35:149–153Google Scholar
  22. Raja N, Goodson M, Smith DG, Rowbury RJ (1991) Decreased DNA damage by acid and increased repair of acid-damaged DNA in acid-habituated Escherichia coli. J Appl Bacteriol 70:507–511PubMedGoogle Scholar
  23. Roe AJ, O'Byrne C, McLaggan D, Booth IR (2002) Inhibition of Escherichia coli growth by acetic acid: a problem with methionine biosynthesis and homocysteine toxicity. Microbiology 148:2215–2222PubMedGoogle Scholar
  24. Russell JB (1992) Another explanation for the toxicity of fermentation acids at low pH: anion accumulation versus uncoupling. J Appl Bacteriol 73:363–370Google Scholar
  25. Russell JB, Diez-Gonzalez F (1998) The effects of fermentation acids on bacterial growth. Adv Microb Physiol 39:205–234PubMedGoogle Scholar
  26. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.Google Scholar
  27. Sharples GJ, Ingleston SM, Lloyd RG (1999) Holliday junction processing in bacteria: insights from the evolutionary conservation of RuvABC, RecG, and RusA. J Bacteriol 181:5543–5550PubMedGoogle Scholar
  28. Singleton MR, Scaife S, Wigley DB (2001) Structural analysis of DNA replication fork reversal by RecG. Cell 107:79–89PubMedGoogle Scholar
  29. Sinha RP (1986) Toxicity of organic acids for repair-deficient strains of Escherichia coli. Appl Environ Microbiol 51:1364–1366PubMedGoogle Scholar
  30. Steiner P, Sauer U (2001) Proteins induced during adaptation of Acetobacter aceti to high acetate concentrations. Appl Environ Microbiol 67:5474–5481CrossRefPubMedGoogle Scholar
  31. Steiner P, Sauer U (2003) Long-term continuous evolution of acetate resistant Acetobacter aceti. Biotechnol Bioeng 83: DOI  10.1002/bit.10741 Google Scholar
  32. Wise A, Brems R, Ramakrishnan V, Villarejo M (1996) Sequences in the −35 region of Escherichia coli rpoS-dependent genes promote transcription by EσS. J Bacteriol 178:2785–2793PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  1. 1.Institute of BiotechnologyETH ZürichZürichSwitzerland

Personalised recommendations