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Research article

Food Microbiology and Biotechnology(FMB)  |  Food Biotechnology

Mass-Based Metabolomic Analysis of Lactobacillus sakei and Its Growth Media at Different Growth Phases

Sang Bong Lee 1, Young Kyoung Rhee 2, Eun-Ji Gu 1, Dong-Wook Kim 1, Gwang-Ju Jang 1, Seong-Hwa Song 1, Jae-In Lee 1, Bo-Min Kim 1, Hyeon-Jeong Lee 1, Hee-Do Hong 2, Chang-Won Cho 2 and Hyun-Jin Kim 1, 3*

1Division of Applied Life Sciences (BK21 Plus), Gyeongsang National University, Jinju 52828, Republic of Korea, 2Korea Food Research Institute, Seongnam 13539, Republic of Korea, 33Department of Food Science & Technology, and Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 52828, Republic of Korea

Received: September 12, 2016; Accepted: February 26, 2017

J. Microbiol. Biotechnol. 2017; 27(5): 925-932

Published May 28, 2017 https://doi.org/10.4014/jmb.1609.09014

Copyright © The Korean Society for Microbiology and Biotechnology.

Abstract

Changes in the metabolite profiles of Lactobacillus sakei and its growth media, based on different culture times (0, 6, 12, and 24 h), were investigated using gas chromatography-mass spectrometry (MS) and liquid chromatography-MS with partial least squares discriminant analysis, in order to understand the growth characteristics of this organism. Cell and media samples of L. sakei were significantly separated on PLS-DA score plots. Cell and media metabolites, including sugars, amino acids, and organic acids, were identified as major metabolites contributing to the difference among samples. The alteration of cell and media metabolites during cell growth was strongly associated with energy production. Glucose, fructose, carnitine, tryptophan, and malic acid in the growth media were used as primary energy sources during the initial growth stage, but after the exhaustion of these energy sources, L. sakei could utilize other sources such as trehalose, citric acid, and lysine in the cell. The change in the levels of these energy sources was inversely similar to the energy production, especially ATP. Based on these identified metabolites, the metabolomic pathway associated with energy production through lactic acid fermentation was proposed. Although further studies are required, these results suggest that MS-based metabolomic analysis might be a useful tool for understanding the growth characteristics of L. sakei, the most important bacterium associated with meat and vegetable fermentation, during growth.

Keywords

Lactobacillus sakei, GC-MS, LC-MS, metabolomics

References

  1. Commane D, Hughes R, Shortt C, Rowland I. 2005. The potential mechanisms involved in the anti-carcinogenic action of probiotics. Mutat. Res. 591: 276-289.
    Pubmed CrossRef
  2. Hammes WP, Tichaczek PS. 1994. The potential of lactic acid bacteria for the production of safe and wholesome food. Z. Lebensm. Unters. Forsch. 198: 193-201.
    Pubmed CrossRef
  3. Leroy F, De Vuyst L. 2004. Lactic acid bacteria as functional starter cultures for the food fermentation industry. Trends Food Sci. Technol. 15: 67-78.
    CrossRef
  4. Hammes WP, Hertel C. 1998. New developments in meat starter cultures. Meat Sci. 49S1: S125-S138.
  5. Leroy F, Verluyten J, De Vuyst L. 2006. Functional meat starter cultures for improved sausage fermentation. Int. J. Food Microbiol. 106: 270-285.
    Pubmed CrossRef
  6. Champomier-Vergès MC, Chaillou S, Cornet M, Zagorec M. 2001. Lactobacillus sakei: recent developments and future prospects. Res. Microbiol. 152: 839-848.
    CrossRef
  7. Dembczynski R, Jankowski T. 2002. Growth characteristics and acidifying activity of Lactobacillus rhamnosus in alginate/starch liquid-core capsules. Enzyme Microb. Technol. 31: 111-115.
    CrossRef
  8. Klaenhammer TR, Kleeman EG. 1981. Growth characteristics, bile sensitivity, and freeze damage in colonial variants of Lactobacillus acidophilus. Appl. Environ. Microbiol. 41: 1461-1467.
    Pubmed PMC
  9. Takahashi H, Kai K, Shinbo Y, Tanaka K, Ohta D, Oshima T, et al. 2008. Metabolomics approach for determining growth specific metabolites based on Fourier transform ion cyclotron resonance mass spectrometry. Anal. Bioanal. Chem. 391: 27692782.
    Pubmed PMC CrossRef
  10. Jin YX, Shi LH, Kawata Y. 2013. Metabolomics-based component profiling of Halomonas sp. KM-1 during dif ferent growth phases in poly(3-hydroxybutyrate) production. Bioresour. Technol. 140: 73-79.
    Pubmed CrossRef
  11. Vidoudez C, Pohnert G. 2012. Comparative metabolomics of the diatom Skeletonema marinoi in different growth phases. Metabolomics 8: 654-669.
    CrossRef
  12. Chen MM, Li AL, Sun MC, Feng Z, Meng XC, Wang Y. 2014. Optimization of the quenching method for metabolomics analysis of Lactobacillus bulgaricus. J. Zhejiang Univ. Sci. B 15: 333-342.
    Pubmed PMC CrossRef
  13. Jäpelt KB, Nielsen NJ, Wiese S, Christensen JH. 2015. Metabolic fingerprinting of Lactobacillus paracasei: a multicriteria evaluation of methods for extraction of intracellular metabolites. Anal. Bioanal. Chem. 407: 6095-6104.
    Pubmed CrossRef
  14. Cui FX, Zhang RM, Liu HQ, Wang YF, Li H. 2015. Metabolic responses to Lactobacillus plantarum contamination or bacteriophage treatment in Saccharomyces cerevisiae using a GC–MS-based metabolomics approach. World J. Microbiol. Biotechnol. 31: 2003-2013.
    Pubmed CrossRef
  15. Zwietering MH, Jongenburger I, Rombouts FM, van’t Riet K. 1990. Modeling of the bacterial growth curve. Appl. Environ. Microbiol. 56: 1875-1881.
    Pubmed PMC
  16. Stentz R, Cornet M, Chaillou S, Zagorec M. 2001. Adaptation of Lactobacillus sakei to meat: a new regulatory mechanism of ribose utilization? Lait 81: 131-138.
    CrossRef
  17. Liu SQ. 2003. Practical implications of lactate and pyruvate metabolism by lactic acid bacteria in food and beverage fermentations. Int. J. Food Microbiol. 83: 115-131.
    CrossRef
  18. Bergmaier D, Champagne CP, Lacroix C. 2003. Exopolysaccharide production during batch cultures with free and immobilized Lactobacillus rhamnosus RW-9595M. J. Appl. Microbiol. 95: 1049-1057.
    Pubmed CrossRef
  19. Vergès MCC, Zuñiga M, Morel-Deville F, Pérez-Martínez G, Zagorec M, Ehrlich SD. 1999. Relationships between arginine degradation, pH and survival in Lactobacillus sakei. FEMS Microbiol. Lett. 180: 297-304.
    CrossRef
  20. Christensen JE, Dudley EG, Pederson JA, Steele JL. 1999. Peptidases and amino acid catabolism in lactic acid bacteria. Antonie Van Leeuwenhoek 76: 217-246.
    Pubmed CrossRef
  21. García-Quintáns N, Blancato VS, Repizo GD, Magni C, López P. 2008. Citrate metabolism and aroma compound production in lactic acid bacteria, pp. 65-88. In Mayo B, López P, Pérez-Martínez G (eds.). Molecular Aspects of Lactic Acid Bacteria for Traditional and New Applications. Research Signpost, Kerala, India.
  22. Van Kranenburg R, Kleerebezem M, Van Hylckama Vlieg J, Ursing BM, Boekhorst J, Smit BA, et al. 2002. Flavour formation from amino acids by lactic acid bacteria: predictions from genome sequence analysis. Int. Dairy J. 12: 111-121.
    CrossRef

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Article

Research article

J. Microbiol. Biotechnol. 2017; 27(5): 925-932

Published online May 28, 2017 https://doi.org/10.4014/jmb.1609.09014

Copyright © The Korean Society for Microbiology and Biotechnology.

Mass-Based Metabolomic Analysis of Lactobacillus sakei and Its Growth Media at Different Growth Phases

Sang Bong Lee 1, Young Kyoung Rhee 2, Eun-Ji Gu 1, Dong-Wook Kim 1, Gwang-Ju Jang 1, Seong-Hwa Song 1, Jae-In Lee 1, Bo-Min Kim 1, Hyeon-Jeong Lee 1, Hee-Do Hong 2, Chang-Won Cho 2 and Hyun-Jin Kim 1, 3*

1Division of Applied Life Sciences (BK21 Plus), Gyeongsang National University, Jinju 52828, Republic of Korea, 2Korea Food Research Institute, Seongnam 13539, Republic of Korea, 33Department of Food Science & Technology, and Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 52828, Republic of Korea

Received: September 12, 2016; Accepted: February 26, 2017

Abstract

Changes in the metabolite profiles of Lactobacillus sakei and its growth media, based on
different culture times (0, 6, 12, and 24 h), were investigated using gas chromatography-mass
spectrometry (MS) and liquid chromatography-MS with partial least squares discriminant
analysis, in order to understand the growth characteristics of this organism. Cell and media
samples of L. sakei were significantly separated on PLS-DA score plots. Cell and media
metabolites, including sugars, amino acids, and organic acids, were identified as major
metabolites contributing to the difference among samples. The alteration of cell and media
metabolites during cell growth was strongly associated with energy production. Glucose,
fructose, carnitine, tryptophan, and malic acid in the growth media were used as primary
energy sources during the initial growth stage, but after the exhaustion of these energy
sources, L. sakei could utilize other sources such as trehalose, citric acid, and lysine in the cell.
The change in the levels of these energy sources was inversely similar to the energy
production, especially ATP. Based on these identified metabolites, the metabolomic pathway
associated with energy production through lactic acid fermentation was proposed. Although
further studies are required, these results suggest that MS-based metabolomic analysis might
be a useful tool for understanding the growth characteristics of L. sakei, the most important
bacterium associated with meat and vegetable fermentation, during growth.

Keywords: Lactobacillus sakei, GC-MS, LC-MS, metabolomics

References

  1. Commane D, Hughes R, Shortt C, Rowland I. 2005. The potential mechanisms involved in the anti-carcinogenic action of probiotics. Mutat. Res. 591: 276-289.
    Pubmed CrossRef
  2. Hammes WP, Tichaczek PS. 1994. The potential of lactic acid bacteria for the production of safe and wholesome food. Z. Lebensm. Unters. Forsch. 198: 193-201.
    Pubmed CrossRef
  3. Leroy F, De Vuyst L. 2004. Lactic acid bacteria as functional starter cultures for the food fermentation industry. Trends Food Sci. Technol. 15: 67-78.
    CrossRef
  4. Hammes WP, Hertel C. 1998. New developments in meat starter cultures. Meat Sci. 49S1: S125-S138.
  5. Leroy F, Verluyten J, De Vuyst L. 2006. Functional meat starter cultures for improved sausage fermentation. Int. J. Food Microbiol. 106: 270-285.
    Pubmed CrossRef
  6. Champomier-Vergès MC, Chaillou S, Cornet M, Zagorec M. 2001. Lactobacillus sakei: recent developments and future prospects. Res. Microbiol. 152: 839-848.
    CrossRef
  7. Dembczynski R, Jankowski T. 2002. Growth characteristics and acidifying activity of Lactobacillus rhamnosus in alginate/starch liquid-core capsules. Enzyme Microb. Technol. 31: 111-115.
    CrossRef
  8. Klaenhammer TR, Kleeman EG. 1981. Growth characteristics, bile sensitivity, and freeze damage in colonial variants of Lactobacillus acidophilus. Appl. Environ. Microbiol. 41: 1461-1467.
    Pubmed KoreaMed
  9. Takahashi H, Kai K, Shinbo Y, Tanaka K, Ohta D, Oshima T, et al. 2008. Metabolomics approach for determining growth specific metabolites based on Fourier transform ion cyclotron resonance mass spectrometry. Anal. Bioanal. Chem. 391: 27692782.
    Pubmed KoreaMed CrossRef
  10. Jin YX, Shi LH, Kawata Y. 2013. Metabolomics-based component profiling of Halomonas sp. KM-1 during dif ferent growth phases in poly(3-hydroxybutyrate) production. Bioresour. Technol. 140: 73-79.
    Pubmed CrossRef
  11. Vidoudez C, Pohnert G. 2012. Comparative metabolomics of the diatom Skeletonema marinoi in different growth phases. Metabolomics 8: 654-669.
    CrossRef
  12. Chen MM, Li AL, Sun MC, Feng Z, Meng XC, Wang Y. 2014. Optimization of the quenching method for metabolomics analysis of Lactobacillus bulgaricus. J. Zhejiang Univ. Sci. B 15: 333-342.
    Pubmed KoreaMed CrossRef
  13. Jäpelt KB, Nielsen NJ, Wiese S, Christensen JH. 2015. Metabolic fingerprinting of Lactobacillus paracasei: a multicriteria evaluation of methods for extraction of intracellular metabolites. Anal. Bioanal. Chem. 407: 6095-6104.
    Pubmed CrossRef
  14. Cui FX, Zhang RM, Liu HQ, Wang YF, Li H. 2015. Metabolic responses to Lactobacillus plantarum contamination or bacteriophage treatment in Saccharomyces cerevisiae using a GC–MS-based metabolomics approach. World J. Microbiol. Biotechnol. 31: 2003-2013.
    Pubmed CrossRef
  15. Zwietering MH, Jongenburger I, Rombouts FM, van’t Riet K. 1990. Modeling of the bacterial growth curve. Appl. Environ. Microbiol. 56: 1875-1881.
    Pubmed KoreaMed
  16. Stentz R, Cornet M, Chaillou S, Zagorec M. 2001. Adaptation of Lactobacillus sakei to meat: a new regulatory mechanism of ribose utilization? Lait 81: 131-138.
    CrossRef
  17. Liu SQ. 2003. Practical implications of lactate and pyruvate metabolism by lactic acid bacteria in food and beverage fermentations. Int. J. Food Microbiol. 83: 115-131.
    CrossRef
  18. Bergmaier D, Champagne CP, Lacroix C. 2003. Exopolysaccharide production during batch cultures with free and immobilized Lactobacillus rhamnosus RW-9595M. J. Appl. Microbiol. 95: 1049-1057.
    Pubmed CrossRef
  19. Vergès MCC, Zuñiga M, Morel-Deville F, Pérez-Martínez G, Zagorec M, Ehrlich SD. 1999. Relationships between arginine degradation, pH and survival in Lactobacillus sakei. FEMS Microbiol. Lett. 180: 297-304.
    CrossRef
  20. Christensen JE, Dudley EG, Pederson JA, Steele JL. 1999. Peptidases and amino acid catabolism in lactic acid bacteria. Antonie Van Leeuwenhoek 76: 217-246.
    Pubmed CrossRef
  21. García-Quintáns N, Blancato VS, Repizo GD, Magni C, López P. 2008. Citrate metabolism and aroma compound production in lactic acid bacteria, pp. 65-88. In Mayo B, López P, Pérez-Martínez G (eds.). Molecular Aspects of Lactic Acid Bacteria for Traditional and New Applications. Research Signpost, Kerala, India.
  22. Van Kranenburg R, Kleerebezem M, Van Hylckama Vlieg J, Ursing BM, Boekhorst J, Smit BA, et al. 2002. Flavour formation from amino acids by lactic acid bacteria: predictions from genome sequence analysis. Int. Dairy J. 12: 111-121.
    CrossRef