Understanding the Starvation Adaptation of Lactobacillus casei through Proteomics

Authors

  • Malik A. Hussain Department of Wine, Food and Molecular Biosciences Room 47, RFH Building, Lincoln University, Christchurch
  • Matthew I. Knight Biosciences Research Division, Department of Primary Industries, Melbourne
  • Margaret L. Britz Tasmanian Institute of Agriculture, Faculty of Science, Engineering and Technology, University of Tasmania, Tasmania 7001

Keywords:

Lactobacilli, Proteomics, Starvation, Adaptation, Metabolic enzymes

Abstract

Food microbes are exposed to several stress conditions in natural environments, which can have an effect on performances. Lactobacilli are highly adaptive group of food microbes which are able to survive in various environmental niches, including ones where preferred nutrients are deficient.  This study was conducted to investigate the adaptation to lactose starvation in Lactobacillus casei using comparative proteomics. One-dimensional sodium dodecyl sulphate-polyacrylamide (1-D SDS PAGE) and two-dimensional electrophoresis (2-DE) were performed on L. casei cells cultivated in a semi-defined medium, with different initial levels of lactose, up to 8 days. Clear visual changes in the 1-D SDS PAGE profiles were seen for cells cultured in 0% lactose. The relative expression of xylulose-5-phosphate phosphoketolase, elongation factor G and DnaK increased in lactose starved cells during stationary phase when compared to the temporal expression of these proteins in cytosolic fraction of cells cultured in 0.2 or 1% lactose. Comparative spot analysis of 2-DE gels showed that 13 proteins were over expressed in lactose starved cells (0% lactose). Of these up-regulated proteins, nine were identified by MALDI-TOF/TOF mass spectrometry with functionalities in protein synthesis, general stress responses and carbohydrate metabolism, where enzymes involved in glycolysis, pyruvate metabolism and the pentose phosphate pathway were up-regulated. These results suggested that proteomic analysis can provide useful information on adaptation of lactobacilli.  Identification of specific protein markers that involve in adaptation to a specific stress factor would be help in selecting strains with better performance in a given application of these beneficial food microbes.

 

References

Adamberg K, Antonsson M, Vogensen FK, Nielsen EW, Kask S, Møller PL, Ardö Y: Fermentation of carbohydrates from cheese sources by non-starter lactic acid bacteria isolated from semi-hard Danish cheese. Int Dairy J 2005, 15:873-882.

Anastasiou R, Leverrier P, Krestas I, Rouault A, Kalantzopoulos G, Boyaval P, Tsakalidou E, Jan G: Changes in protein synthesis during thermal adaptation of Propionibacterium freudenreichii subsp. shermanii. Int J Food Microbiol 2006, 108:301-314.

Bernhardt J, Völker U, Völker A, Antelmann H, Schmid R, Mach H, Hecker M: Specific and general stress proteins in Bacillus subtilis – a two-dimensional electrophoretic study. Microbiol 1997, 143:999-1017.

Bourne HR, Sanders DA, McCormick F: The GTPase superfamily: conserved structure and molecular mechanism. Nature 1991, 349:117-127.

Broadbent JR, Lin C: Effect of heat shock or cold shock treatment on the resistance of Lactococcus lactis to freezing and lyophilization. Cryobiology 1999, 39:88–102.

Hamon E, Horvatovich P, Izquierdo E, Bringel F, Marchioni E, Aoudé-Werner D, Ennahar S: Comparative proteomic analysis of Lactobacillus plantarum for the identification of key proteins in bile tolerance. BMC Microbiol 2011, 11: 63

Champomier-Vergès M, Maguin E, Mistou M, Anglade P, Chich J: Lactic acid bacteria and proteomics: current knowledge and perspectives. J Chromat B 2002, 771:329-342.

Chandry PS, Moore SC, Davidson BE, Hillier AJ: Investigation of the microbial ecology of maturing cheese by PCR and PFGE. Aust J Dairy Technol 1998, 53:17.

Cohen DPA, Renes J, Bouwman FG, Zoetendal EG, Mariman E, de Vos WM, Vaughan EE: Proteomic analysis of log to stationary growth phase Lactobacillus plantarum cells and a 2-DE database. Proteomics 2006, 6:6485-6493.

De Angelis M, Bini L, Pallini V, Cocconcelli PS, Gobbetti M: The acid-stress response in Lactobacillus sanfranciscensis CB1. Microbiology 2001, 147:1863-1873.

De Angelis M, Di Cagno R, Huet C, Crecchio C, Fox PF, Gobbetti M: Heat shock response in Lactobacillus plantarum. Appl Environ Microbiol 2004, 70:1336-1346.

De Angelis M, Gobbetti M: Environmental stress responses in Lactobacillus: A review. Proteomics 2004, 4:106-122.

Fernandez A, Ogawa J, Penaud S, Boudebbouze S, Ehrlich D, van de Guchte M, Maguin E: Rerouting of pyruvate metabolism during acid adaptation in Lactobacillus bulgaricus. Proteomics 2008, 8:3154-63.

Frece J, Kos B, Svetec IK, Zgaga Z, Mrsa V, Suskovic J: Importance of S-layer proteins in probiotic activity of Lactobacillus acidophilus M92. J Appl Microbiol 2005, 98:285-292.

Gagnaire V, Piot M, Camier B, Vissers JPC, Jan G, Léonil J: Survey of bacterial proteins released in cheese: a proteomic approach. Int J Food Microbiol 2004, 94:185-201.

Ganesan B, Stuart MR, Weimer BC: Carbohydrate starvation causes a metabolically active but nonculturable state in Lactococcus lactis. Appl Environ Microbiol 2007, 73:2498-2512.

Giard JC, Laplace JM, Rince A, Pichereau V, Benachour A, Leboeuf C, Flahaut S, Auffray Y, Hartke A: The stress proteome of Enterococcus faecalis. Electrophoresis 2001, 22:2947-2954.

Gottesman S, Wickner S, Maurizi MR: Protein quality control: triage by chaperones and proteases. Genes Dev 1997, 11:815-823.

Gouesbet G, Jan G, Boyaval P: Two-dimensional electrophoresis study of Lactobacillus delbrueckii subsp. bulgaricus thermotolerance. Appl Environ Microbiol 2002, 68:1055–1063.

Goulhena F, Grenier D, Mayrand D: Stress response in Actinobacillus actinomycetemcomitans: induction of general and specific stress proteins. Res Microbiol 2003, 154:43-48.

Graumann P, Schroder K, Schmid R, Marahiel MA: Cold shock stress-induced proteins in Bacillus subtilis. J Bacteriol 1996, 178:4611-4619.

Gummalla S, Broadbent JR: Trptophan catabolism by Lactobacillus casei and Lactobacillus helveticus cheese flavor adjuncts. J Dairy Sci 1999, 82:2070-2077.

Hartke A, Bouché S, Giard J, Benachour A, Boutibonnes P, Auffray Y: The lactic acid stress response of Lactococcus lactis subsp. lactis. Curr Microbiol 1996, 33:194-199.

Hendrick JP, Hartl F: Molecular chaperone functions of heat-Shock proteins. Annu Rev Biochem 1993, 62:349-384.

Hosseini Nezhad M, Stenzel DJ, Britz ML: Phenomic and proteomic characterization of Lactobacillus casei in response to acid stress. New Biotechnol 2009, 25:347.

Hussain MA, Britz ML. Analysis of long-term survival of NSLAB strains in a semi-defined liquid medium. Aust J Dairy Technol 2006, 61:217.

Hussain MA, Knight MI, Britz ML: Proteomic analysis of lactose starved Lactobacillus casei during stationary growth phase. J Appl Microbiol 2010, 106:764-773.

Hussain MA, Rouch DA, Britz ML: Biochemistry of NSLAB isolate Lb. casei GCRL163: production of metabolites by stationary-phase cultures. Int Dairy J 2009, 19:12-21.

Jakava-Viljanen M, Åvall-Jääskeläinen S, Messner P, Sleytr UB, Palva A: Isolation of three new surface layer protein genes (slp) from Lactobacillus brevis ATCC 14869 and characterization of the change in their expression under aerated and anaerobic conditions. J Bacteriol 2002, 184:6786-6795.

Kilstrup M, Jacobsen S, Hammer K, Vogensen FK.: Induction of heat shock proteins DnaK, GroEL, and GroES by salt stress in Lactococcus lactis. Appl Environ Microbiol 1997, 63:1826–1837.

Kim WS, Park JH, Ren J, Su P, Dunn NW: Survival response and rearrangement of plasmid DNA of Lactococcus lactis during long-term starvation. Appl Environ Microbiol 2001, 10:4594–4602.

Koebmann BJ, Andersen HW, Solem C, Jensen PR: Experimental determination of control of glycolysis in Lactococcus lactis. Antonie van Leeuwenhoek 2002, 82:237–248.

Laemmli UK.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970, 15:680-685.

Lithgow JK, Hayhurst EJ, Cohen G, Aharonowitz Y, Foster SJ: Role of a cysteine synthase in Staphylococcus aureus. J Bacteriol 2004, 186:1579-1590.

Lorca GL, De Valdez F: A low-pH-inducible, stationary-phase acid tolerance response in Lactobacillus acidophilus CRL 639. Curr Microbiol 2001, 42:21–25.

Lorca GL, de Valdez GF, Ljungh A: Characterization of the proteinsynthesis dependent adaptive acid tolerance response in Lactobacillus acidophilus. J Mol Microbiol Biotechnol 2002, 4:525–532.

Macario AJ, Malz M, Conway M: Evolution of assisted protein folding: the distribution of the main chaperoning systems within the phylogenetic domain archaea. Front Biosci 2004, 9:1318-1332.

Mallidis C, Galiatatou P, Taoukis PS, Tassou C: The kinetic evaluation of the use of high hydrostatic pressure to destroy Lactobacillus plantarum and Lactobacillus brevis. Int J Food Microbiol 2003, 38:579-585.

McSweeney PL: Biochemistry of cheese ripening. Int J Dairy Technol 2004, 57:127-144.

Nyström T: Global systems approach to the physiology of the starved cell. In: Kjelleberg S (ed) Starvation in Bacteria . New York: Plenum; 1993, pp 129-150.

Nyström T: Stationary-phase physiology. Annu Rev Microbiol 2004, 58:161-181.

O’Farrell PH: High resolution two-dimensional electrophoresis of proteins. J Biol Chem 1975, 250:4007-4021.

Perkins DN, Pappin DJC, Creasy DM, Cottrell JS: Probability-based protein identification by searching sequence databases using mass spectrometry data. Electrophoresis 1999, 20:3551–3567.

Pessione E, Mazzoli R, Giuffrida MG, Lamberti C, Gracia-Moruno E, Barello C, Conti A, Giunta C: A proteomic approach to studying biogenic amine producing lactic acid bacteria. Proteomics 2005, 5:687-689.

Piuri M, Sanchez-Rivas C, Ruzal SM: Cell wall modifications during osmotic stress in Lactobacillus casei. J Appl Microbiol 2005, 98:84-95.

Posthuma CC, Bader R, Engelmann R, Postma PW, Hengstenberg W, Pouwels PH: Expression of the xylulose 5-phosphate phosphoketolase gene, xpkA, from Lactobacillus pentosus MD363 is induced by sugars that are fermented via the phosphoketolase pathway and is repressed by glucose mediated by ccpA and the mannose phosphoenolpyruvate phosphotransferase system. Appl Environ Microbiol 2002, 68:831-837.

Rabilloud T, Carpentier G, Tarroux P: Improvement and simplification of low background silver staining proteins by using sodium dithonite. Electrophoresis 1988, 9:288-291.

Rabilloud T, Strub JM, Luche S, van Dorsselaer A, Lunardi J: A comparison between Sypro Ruby and ruthenium II tris (bathophenanthroline disulphonate) as fluorescent stains for protein detection in gels. Proteomics 2001, 1:699-704.

Rallu F, Gruss A, Ehrlich SD, Maguin E: Acid- and multistress-resistant mutants of Lactococcus lactis: identification of intracellular stress signals. Mol Microbiol 2000, 35:517–528.

Rouch DA, Hillier AJ, Britz ML: NSLAB in cheddar: a stressful life. Aust J Dairy Technol 2002, 57:107.

Sánchez B, Champomier-Vergès C-C, Anglade P, Baraige F, de los Reyes-Gavilán CG, Margolles A, Zagorec M: Proteomic analysis of global changes in protein expression during bile salt exposure of Bifidobacterium longum NCIMB 8809. J Bacteriol 2005, 187:5799-5808.

Sauvageot N, Beaufils S, Maze A, Deutscher J, Hartke A: Cloning and characterization of a gene encoding cold-shock protein in Lactobacillus casei. FEMS Microbiol Lett 2006, 254:55-62.

Schär-Zammaretti P, Dillman M-L, D’Amico N, Affolter M, Ubbink M: Influence of fermentation medium composition on physiochemical surface properties of Lactobacillus acidophilus. Appl Environ Microbiol 2005, 71:8165-8173.

Scheyhing CH, Hormann S, Ehrmann MA, Vogel RF: Barotolerance is inducible by preincubation under hydrostatic pressure, cold-, osmotic- and acid-stress conditions in Lactobacillus sanfranciscensis DSM 20451. Lett Appl Microbiol 2004, 39:284-289.

Serrano LM, Molenaar D, Wels M, Teusink B, Bron PA, de Vos WM, Smid E: Thioredoxin reductase is a key factor in the oxidative stress response of Lactobacillus plantarum WCFS1. Microb Cell Fact 2007, 6:29.

Shevchenko A, Wilm M, Vorm O, Mann M. Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem 1997, 68:850-858.

Silva J, Carvalho AS, Ferreira R, Vitorino R, Amado F, Domingues P, Teixeira P, Gibbs PA: Effect of the pH of growth on the survival of Lactobacillus delbrueckii subsp. bulgaricus to stress conditions during spray-drying. J Appl Microbiol 2005, 98:775–782.

Smit BA, Engels WJM, Wouters JTM, Smit G: Diversity of L-leucine catabolism in various microorganisms involved in dairy fermentations, and identification of the rate-controlling step in the formation of the potent flavour component 3-methylbutanal. Appl Microbiol Biotechnol 2004, 64:396-402.

Spano G, Capozzi V, Vernile A, Massa S: Cloning, molecular characterization and expression analysis of two small heat shock genes isolated from wine Lactobacillus plantarum. J Appl Microbiol 2004, 97:774-782.

Spector MP, Foster JW: Starvation-stress response (SSR) of Salmonella typhimurium. In: Kjelleberg S (ed) Starvation in Bacteria. New York: Plenum; 1993, pp 201-224.

Sprang SR: G protein mechanisms: insights from structural analysis. Annu Rev Biochem 1997, 66:639-678.

Steiner P, Sauer U: Proteins induced during adaptation of Acetobacter aceti to high acetate concentrations. Appl Environ Microbiol 2001, 67:5474-5481.

Stuart MR, Chou LS, Weimer BC: Influence of carbohydrate starvation and arginine on culturability and amino acid utilization of Lactococcus lactis subsp. lactis. Appl Environ Microbiol 1999, 65:665-673.

VanBogelen RA, Kelly PM, Neidhardt FC: Gene–protein database of Escherichia coli K-12: edition 3. Electrophoresis 1990, 11:1131-1166.

Alcántara C, Zúñiga M: Proteomic and transcriptomic analysis of the response to bile stress of L. casei BL23. Microbiology 2012,158:1206-1218.

Waldron DS: Effect of lactose concentration on the quality of cheddar cheese. MSc Thesis. University College Cork, .National University of Ireland. 1997.

Wickner S, Gottesman S, Skowyra D, Hoskins J, Mckenney K, Maurizi MR: A molecular chaperone, Clpa, functions like DnaK and DnaJ. Biochemistry 1994, 91:12218-12222.

Wilkins JC, Homer KA, Beighton D: Analysis of Streptococcus mutans proteins modulated by culture under acidic conditions. Appl Environ Microbiol 2002, 68:2382-2390.

Willemoës M, Kilstrup M, Roepstorff P, Hammer K: Proteome analysis of a Lactococcus lactis strain overexpressing gapA suggests that the gene product is an auxiliary glyceraldehyde 3-phosphate dehydrogenase. Proteomics 2002, 2:1041-1046.

Williams AG, Noble J, Banks JM: Catabolism of amino acids by lactic acid bacteria isolated from Cheddar cheese. Int Dairy J 2001, 11:203-215.

Wouters JA, Hamphuis KH, Kuipers OP, De Vos WM, Abee T: Changes in glycolytic activity of Lactococcus lactis induced by low temperature. Appl Environ Microbiol 2000, 66:3686-3691.

Wu R, Zhang WY, Sun T, Wu JR, Yue X, Meng H, Zhang HP: Proteomic analysis of responses of a new probiotic bacterium Lactobacillus casei Zhang to low acid stress. Int J Food Microbio. 2011, 147:181-187.

Yan JX, Wait R, Berkelman T, Harry R, Westbrook J, Wheeler C, Dunn M: A modified silver staining protocol for visualization of proteins compatible with matrix-assisted laser desorption/ionisation and electrospray ionisation–mass spectrometry. Electrophoresis 2000, 21:3666-3672.

Butorac A, Dodig I, BaÄun-Družina V, Tishbee A, MrvÄić J, et al (2013) The effect of starvation stress on Lactobacillus brevis L62 protein profile determined by de novo sequencing in positive and negative mass spectrometry ion mode. Rapid Comm Mass Spect 27:1045.

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Published

2013-12-13

How to Cite

Hussain, M. A., Knight, M. I., & Britz, M. L. (2013). Understanding the Starvation Adaptation of Lactobacillus casei through Proteomics. Asian Journal of Agriculture and Food Sciences, 1(5). Retrieved from https://www.ajouronline.com/index.php/AJAFS/article/view/578