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Influence of polysorbate 80 and cyclopropane fatty acid synthase activity on lactic acid production by Lactobacillus casei ATCC 334 at low pH

  • J. R. BroadbentEmail author
  • T. S. Oberg
  • J. E. Hughes
  • R. E. Ward
  • C. Brighton
  • D. L. Welker
  • J. L. Steele
Fermentation, Cell Culture and Bioengineering

Abstract

Lactic acid is an important industrial chemical commonly produced through microbial fermentation. The efficiency of acid extraction is increased at or below the acid’s pKa (pH 3.86), so there is interest in factors that allow for a reduced fermentation pH. We explored the role of cyclopropane synthase (Cfa) and polysorbate (Tween) 80 on acid production and membrane lipid composition in Lactobacillus casei ATCC 334 at low pH. Cells from wild-type and an ATCC 334 cfa knockout mutant were incubated in APT broth medium containing 3 % glucose plus 0.02 or 0.2 % Tween 80. The cultures were allowed to acidify the medium until it reached a target pH (4.5, 4.0, or 3.8), and then the pH was maintained by automatic addition of NH4OH. Cells were collected at the midpoint of the fermentation for membrane lipid analysis, and media samples were analyzed for lactic and acetic acids when acid production had ceased. There were no significant differences in the quantity of lactic acid produced at different pH values by wild-type or mutant cells grown in APT, but the rate of acid production was reduced as pH declined. APT supplementation with 0.2 % Tween 80 significantly increased the amount of lactic acid produced by wild-type cells at pH 3.8, and the rate of acid production was modestly improved. This effect was not observed with the cfa mutant, which indicated Cfa activity and Tween 80 supplementation were each involved in the significant increase in lactic acid yield observed with wild-type L. casei at pH 3.8.

Keywords

Lactobacillus Membrane Fermentation Lactic acid 

Notes

Acknowledgments

This project was supported by National Research Initiative Competitive Grant no. 2011-67009-30043 from the USDA National Institute of Food and Agriculture, Program, and by the Utah Agricultural Experiment Station. This communication is approved as UAES Journal Paper Number 8589. Peggy Steele, a member of Dr. Steele’s family, is employed by Dupont Inc., a supplier of bacterial cultures to the food industry.

References

  1. 1.
    Baniel AM, Aharon EM, Mizrahi J, Hazan B, Fisher RR, Kolstad JJ, Stewart BF (2000) Process for isolating lactic acid. US Patent 6,087,532Google Scholar
  2. 2.
    Booth IR (1985) Regulation of cytoplasmic pH in bacteria. Microbiol Rev 49:359–378PubMedCentralPubMedGoogle Scholar
  3. 3.
    Broadbent JR, Larsen RL, Diebold V, Steele JL (2010) Physiological and transcriptional response of Lactobacillus casei ATCC 334 to acid stress. J Bacteriol 192:2445–2458PubMedCentralPubMedCrossRefGoogle Scholar
  4. 4.
    Carpenter CE, Broadbent JR (2009) External concentration of organic acid anions and pH: key independent variables for studying how organic acids inhibit growth of bacteria in mildly acidic foods. J Food Sci 74:R12–R15PubMedCrossRefGoogle Scholar
  5. 5.
    Corcoran BM, Stanton C, Fitzgerald GF, Ross RP (2007) Growth of probiotic lactobacilli in the presence of oleic acid enhances subsequent survival in gastric juice. Microbiol 153:291–299CrossRefGoogle Scholar
  6. 6.
    Cotter PD, Hill C (2003) Surviving the acid test: responses of Gram-positive bacteria to low pH. Microbiol Mol Biol Rev 67:429–453PubMedCentralPubMedCrossRefGoogle Scholar
  7. 7.
    Demirci A, Pometto AL III, Lee B, Hinz PN (1998) Media evaluation of lactic acid repeated-batch fermentation with Lactobacillus plantarum and Lactobacillus casei subsp. rhamnosus. J Agric Food Chem 46:4771–4774CrossRefGoogle Scholar
  8. 8.
    Denich TJ, Beaudette LA, Lee H, Trevors JT (2003) Effect of selected environmental and physico-chemical factors on bacterial cytoplasmic membranes. J Microbiol Methods 52:149–182PubMedCrossRefGoogle Scholar
  9. 9.
    Fozo EM, Kajfasz JK, Quivey RG Jr (2004) Low pH-induced membrane fatty acid alterations in oral bacteria. FEMS Microbiol Lett 238:291–295PubMedCrossRefGoogle Scholar
  10. 10.
    Grogan DW, Cronan JE Jr (1997) Cyclopropane ring formation in membrane lipids of bacteria. Microbiol Mol Biol Rev 61:429–441PubMedCentralPubMedGoogle Scholar
  11. 11.
    Hayter A (2007) Probability and statistics for engineers and scientists. Thompson Brooks/Cole, BelmontGoogle Scholar
  12. 12.
    Hutkins RW, Nannen NL (1993) pH homeostasis in lactic acid bacteria. J Dairy Sci 76:2354–2365CrossRefGoogle Scholar
  13. 13.
    Johnsson T, Nikkila P, Toivonen L, Rosenqvist H, Laakso S (1995) Cellular fatty acid profiles of Lactobacillus and Lactococcus strains in relation to the oleic acid content of the cultivation medium. Appl Environ Microbiol 61:4497–4499PubMedCentralPubMedGoogle Scholar
  14. 14.
    Kandler O, Weiss N (1986) Genus Lactobacillus. In: Sneath PHA, Mair NS, Sharpe ME, Holt JG (eds) Bergey’s manual of systematic bacteriology, vol 2, 9th edn. Williams and Wilkins, Baltimore, pp 1208–1234Google Scholar
  15. 15.
    Kashket ER (1987) Bioenergetics of lactic acid bacteria: cytoplasmic pH and osmotolerance. FEMS Microbiol Rev 46:233–244CrossRefGoogle Scholar
  16. 16.
    Kristich CJ, Chandler JR, Dunny GM (2007) Development of a host-genotype-independent counterselectable marker and a high-frequency conjugative delivery system and their use in genetic analysis of Enterococcus faecalis. Plasmid 57:131–144PubMedCentralPubMedCrossRefGoogle Scholar
  17. 17.
    Lambert RJ, Stratford M (1999) Weak-acid preservatives: modeling microbial inhibition and response. J Appl Microbiol 86:157–164PubMedCrossRefGoogle Scholar
  18. 18.
    Leenhouts K, Buist G, Bolhuis A, ten Berge A, Kiel J, Mierau I, Dabrowska M, Venema G, Kok J (1996) A general system for generating unlabelled gene replacements in bacterial chromosomes. Mol Gen Genet 253:217–224PubMedCrossRefGoogle Scholar
  19. 19.
    Mykytczuk NCS, Trevors JT, Leduc LG, Ferroni GD (2007) Fluorescence polarization in studies of bacterial cytoplasmic membrane fluidity under environmental stress. Prog Biophys Mol Biol 95:60–82PubMedCrossRefGoogle Scholar
  20. 20.
    Narayanan N, Roychoudhury PK, Srivastava A (2004) L (+) lactic acid fermentation and its product polymerization. Electronic J Biotechnol 7:167–179Google Scholar
  21. 21.
    Oberg TS, Ward RE, Steele JL, Broadbent JR (2012) Identification of plasmalogens in the cytoplasmic membrane of Bifidobacterium animalis subsp. lactis. Appl Environ Microbiol 78:880–884PubMedCentralPubMedCrossRefGoogle Scholar
  22. 22.
    Russell JB (1991) Resistance of Streptococcus bovis to acetic acid at low pH: relationship between intracellular pH and anion accumulation. Appl Environ Microbiol 57:255–259PubMedCentralPubMedGoogle Scholar
  23. 23.
    Russell JB (1992) Another explanation for the toxicity of fermentation acids at low pH; anion accumulation verses uncoupling. J Appl Microbiol 73:363–370Google Scholar
  24. 24.
    Russell NJ (1984) Mechanisms of thermal adaptation in bacteria, blueprints for survival. Trends Biochem Sci 3:108–112CrossRefGoogle Scholar
  25. 25.
    Sasser M (1990) Identification of bacteria by gas chromatography of cellular fatty acids. Tech note 101. Midi Inc., Newark, DE. http://www.microbialid.com/PDF/TechNote_101.pdf
  26. 26.
    Shabala L, Ross T (2008) Cyclopropane fatty acids improve Escherichia coli survival in acidified minimal media by reducing membrane permeability to H+ and enhanced ability to extrude H+. Res Microbiol 159:458–461PubMedCrossRefGoogle Scholar
  27. 27.
    Suutari M, Laakso S (1992) Temperature adaptation in Lactobacillus fermentum: interconversions of oleic, vaccenic and dihydrosterulic acids. J Gen Microbiol 138:445–450PubMedCrossRefGoogle Scholar
  28. 28.
    Varadarajan S, Miller DJ (1999) Catalytic upgrading of fermentation-derived organic acids. Biotechnol Prog 15:845–854PubMedCrossRefGoogle Scholar
  29. 29.
    Vigh L, Escribá P, Sonnleitner A, Sonnleitner M, Piotto S, Maresca B, Horváth I, Harwood J (2005) The significance of lipid composition for membrane activity: new concepts and ways of assessing function. Prog Lipid Res 44:303–344PubMedCrossRefGoogle Scholar
  30. 30.
    Vijayakumar J, Aravindan R, Viruthagiri T (2008) Recent trends in the production, purification and application of lactic acid. Chem Biochem Eng Q 22:245–264Google Scholar
  31. 31.
    Vink ETH, Glassner DA, Kolstad JJ, Wooley RJ, O’Connor RP (2007) The eco-profiles for current and near-future NatureWorks polylactide (PLA) production. Ind Biotechnol 3:58–81CrossRefGoogle Scholar
  32. 32.
    Wee YJ, Kim JN, Ryu HW (2006) Biotechnological production of lactic acid and its recent applications. Food Technol Biotechnol 44:163–172Google Scholar
  33. 33.
    Wu C, Zhang J, Wang M, Du G, Chen J (2012) Lactobacillus casei combats acid stress by maintaining cell membrane functionality. J Ind Microbiol Biotechnol 39:1031–1039PubMedCrossRefGoogle Scholar
  34. 34.
    Zhang Y-M, Rock CO (2008) Membrane lipid homeostasis in bacteria. Nat Rev Micro 6:222–233CrossRefGoogle Scholar
  35. 35.
    Zhang Y-M, Rock CO (2009) Transcriptional regulation in bacterial membrane lipid synthesis. J Lipid Res 50(Suppl.):S115–S119Google Scholar

Copyright information

© Society for Industrial Microbiology and Biotechnology 2013

Authors and Affiliations

  • J. R. Broadbent
    • 1
    Email author
  • T. S. Oberg
    • 1
  • J. E. Hughes
    • 2
  • R. E. Ward
    • 1
  • C. Brighton
    • 1
  • D. L. Welker
    • 2
  • J. L. Steele
    • 3
  1. 1.Department of Nutrition, Dietetics, and Food ScienceUtah State UniversityLoganUSA
  2. 2.Department of BiologyUtah State UniversityLoganUSA
  3. 3.Department of Food ScienceUniversity of WisconsinMadisonUSA

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