Hostname: page-component-cd9895bd7-fscjk Total loading time: 0 Render date: 2024-12-28T03:32:09.921Z Has data issue: false hasContentIssue false
  • English
  • Français

Screening for antimicrobial and proteolytic activities of lactic acid bacteria isolated from cow, buffalo and goat milk and cheeses marketed in the southeast region of Brazil

Published online by Cambridge University Press:  26 November 2015

Fabricio L Tulini ,
Nolwenn Hymery ,
Thomas Haertlé ,
Gwenaelle Le Blay  and
Elaine C P De Martinis
Fabricio L Tulini
Affiliation:
Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo – Ribeirão Preto, Brazil
Nolwenn Hymery
Affiliation:
Université de Brest, EA3882, Laboratoire Universitaire de Biodiversité et d'Ecologie Microbienne, SFR ScInBioS, ESIAB, Technopôle de Brest Iroise, 29280 Plouzané, France
Thomas Haertlé
Affiliation:
Institut National de la Recherche Agronomique (INRA) Angers-Nantes, BIA-FIPL – Nantes, France
Gwenaelle Le Blay
Affiliation:
Université de Brest, UMR UBO, CNRS, IFREMER 6197, Laboratoire de Microbiologie des Environnements Extrêmes, IUEM, Technopôle de Brest Iroise, 29280 Plouzané, France
Elaine C P De Martinis*
Affiliation:
Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo – Ribeirão Preto, Brazil
*
*For correspondence; e-mail: edemarti@usp.br
Get access Rights & Permissions [Opens in a new window]

Abstract

Lactic acid bacteria (LAB) can be isolated from different sources such as milk and cheese, and the lipolytic, proteolytic and glycolytic enzymes of LAB are important in cheese preservation and in flavour production. Moreover, LAB produce several antimicrobial compounds which make these bacteria interesting for food biopreservation. These characteristics stimulate the search of new strains with technological potential. From 156 milk and cheese samples from cow, buffalo and goat, 815 isolates were obtained on selective agars for LAB. Pure cultures were evaluated for antimicrobial activities by agar antagonism tests and for proteolytic activity on milk proteins by cultivation on agar plates. The most proteolytic isolates were also tested by cultivation in skim milk followed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the fermented milk. Among the 815 tested isolates, three of them identified as Streptococcus uberis (strains FT86, FT126 and FT190) were bacteriocin producers, whereas four other ones identified as Weissella confusa FT424, W. hellenica FT476, Leuconostoc citreum FT671 and Lactobacillus plantarum FT723 showed high antifungal activity in preliminary assays. Complementary analyses showed that the most antifungal strain was L. plantarum FT723 that inhibited Penicillium expansum in modified MRS agar (De Man, Rogosa, Sharpe, without acetate) and fermented milk model, however no inhibition was observed against Yarrowia lipolytica. The proteolytic capacities of three highly proteolytic isolates identified as Enterococcus faecalis (strains FT132 and FT522) and Lactobacillus paracasei FT700 were confirmed by SDS–PAGE, as visualized by the digestion of caseins and whey proteins (β-lactoglobulin and α-lactalbumin). These results suggest potential applications of these isolates or their activities (proteolytic activity or production of antimicrobials) in dairy foods production.

Keywords

Lactic acid bacteriadairyantimicrobialsproteinaseidentification
Type
Research Article
Information
Journal of Dairy Research , Volume 83 , Issue 1 , February 2016 , pp. 115 - 124
Copyright
Copyright © Proprietors of Journal of Dairy Research 2015 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Ahmadova, A, Dimov, S, Ivanova, E, Choiset, Y, Chobert, J-M, Kuliev, A & Haertlé, T 2011 Proteolytic activities and safety of use of Enterococci strains isolated from traditional Azerbaijani dairy products. European Food Research and Technology 233 131–140CrossRefGoogle Scholar
Axelsson, LT 1993 Lactic acid bacteria: classification and physiology. In Lactic Acid Bacteria, pp. 1–63 (Eds Salminenand, S & Wright, AT). New York: Marcel DekkerGoogle Scholar
Baek, E, Kim, H, Choi, H, Yoon, S & Kim, J 2012 Antifungal activity of Leuconostoc citreum and Weissella confusa in rice cakes. Journal of Microbiology 50 842–848CrossRefGoogle ScholarPubMed
Berthier, F & Ehrlich, SD 1998 Rapid species identification within two groups of closely related lactobacilli using PCR primers that target the 16S/23S rRNA spacer region. FEMS Microbiology Letters 161 97–106CrossRefGoogle ScholarPubMed
Chanos, P & Williams, DR 2011 Anti-Listeria bacteriocin-producing bacteria from raw ewe's milk in northern Greece. Journal of Applied Microbiology 110 757–768CrossRefGoogle ScholarPubMed
Crowley, S, Mahony, J & Van Sinderen, D 2013 Current perspectives on antifungal lactic acid bacteria as natural bio-preservatives. Trends in Food Science and Technology 33 93–109CrossRefGoogle Scholar
Cuiv, PO, Klaassens, ES, Smith, WJ, Mondot, S, Durkin, AS, Harkins, DM, Foster, L, McCorrinson, J, Torralba, M, Nelson, KE & Morrison, M 2013 Draft genome sequence of Enterococcus faecalis PC1.1, a candidate probiotic strain isolated from human feces. Genome Announcements 1 1–2CrossRefGoogle Scholar
Delavenne, E, Mounier, J, Déniel, F, Barbier, G & Le Blay, G 2012 Biodiversity of antifungal lactic acid bacteria isolated from raw milk samples from cow, ewe and goat over one-year period. International Journal of Food Microbiology 155 185–190CrossRefGoogle ScholarPubMed
Delavenne, E, Ismail, R, Pawtowski, A, Mounier, J, Barbier, G & Le Blay, G 2013 Assessment of lactobacilli strains as yogurt bioprotective cultures. Food Control 30 206–213CrossRefGoogle Scholar
De Man, JD, Rogosa, M & Sharpe, ME 1960 A medium for the cultivation of Lactobacilli. Journal of Applied Bacteriology 23 130–135CrossRefGoogle Scholar
Douillard, FP, Kant, R, Ritari, J, Paulin, L, Airi, P & De Vos, WM 2013 Comparative genome analysis of Lactobacillus casei strains isolated from Actimel and Yakult products reveals marked similarities and points to a common origin. Microbial Biotechnology 6 576–587CrossRefGoogle ScholarPubMed
Dutka-Malen, S, Evens, S & Couvarlin, P 1995 Detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant Enterococci by PCR. Journal of Clinical Microbiology 33 24–27CrossRefGoogle Scholar
El-Ghaish, S, Dalgalarrondo, M, Choiset, Y, Sitohy, M, Ivanova, I, Haertlé, T & Chobert, J-M 2010 Screening of strains of lactococci isolated from Egyptian dairy products for their proteolytic activity. Food Chemistry 120 758–764CrossRefGoogle Scholar
Food and Drug Administration 2012 Summary of data concerning the safety and GRAS determination of food ferment solutions for use as a food ingredient. http://www.fda.gov/ucm/groups/fdagov-public/@fdagov-foods-gen/documents/document/ucm273648.pdf (24 February 2015)Google Scholar
Fritzenwanker, M, Kuenne, C, Billion, A, Hain, T, Zimmermann, K, Goesmann, A, Chakraborty, T & Domann, E 2013 Complete genome sequence of the probiotic Enterococcus faecalis Symbioflor 1 clone DSM 16431. Genome Announcements 1 1–2CrossRefGoogle ScholarPubMed
Galvez, A, Abriouel, H, Lopez, RL & Ben Omar, N 2007 Bacteriocin-based strategies for food biopreservation. International Journal of Food Microbiology 120 51–70CrossRefGoogle ScholarPubMed
Guinane, CM, Cotter, PD, Hill, C & Ross, RP 2005 Microbial solutions to microbial problems; lactococcal bacteriocins for the control of undesirable biota in food. Journal of Applied Microbiology 98 1316–1325CrossRefGoogle ScholarPubMed
Hartemink, R, Van Laere, KM & Rombouts, FM 1997 Growth of enterobacteria on fructo-oligosaccharides. Journal of Applied Microbiology 83 367–374CrossRefGoogle ScholarPubMed
Harwood, VJ, Brownell, M, Perusek, W & Whitlock, JE 2001 Vancomycin-resistant Enterococcus spp. isolated from wastewater and chicken feces in the United States. Applied and Environmental Microbiology 67 4930–4933CrossRefGoogle ScholarPubMed
Jackson, CR, Fedorka-Cray, PJ & Barrett, JB 2004 Use of a genus- and species-specific multiplex PCR for identification of Enterococci. Journal of Clinical Microbiology 42 3558–3565CrossRefGoogle ScholarPubMed
Lane, DJ 2001 16S/23S rRNA sequencing. In Nucleic Acid Techniques in Bacterial Systematics, pp. 115–175 (Eds Stackebrandt, E & Goodfellow, M). New York, USA: John Wiley & SonsGoogle Scholar
Lee, HJ, Park, SY & Kim, J 2000 Multiplex PCR-based detection and identification of Leuconostoc species. FEMS Microbiology Letters 193 243–247CrossRefGoogle ScholarPubMed
Lee, KW, Park, JY, Jeong, HR, Heo, HJ, Han, NS & Kim, JH 2012 Probiotic properties of Weissella strains isolated from human faeces. Anaerobe 18 96–102CrossRefGoogle ScholarPubMed
Leigh, JA 1999 Streptococcus uberis: a permanent barrier to the control of bovine mastitis? Veterinary Journal 157 225–238CrossRefGoogle Scholar
Lewus, CB, Kaiser, A & Montville, TJ 1991 Inhibition of food-borne bacteria pathogens by bacteriocins from lactic acid bacteria isolated from meat. Applied and Environmental Microbiology 57 1683–1688CrossRefGoogle ScholarPubMed
Nero, LA, Mattos, MR, Beloti, V, Barros, MAF, Ortolani, MBT & Franco, BDGM 2009 Autochthonous microbiota of raw milk with antagonistic activity against Listeria monocytogenes and Salmonella Enteritidis. Journal of Food Safety 29 261–270CrossRefGoogle Scholar
Nilsson, L, Ng, YY, Christiansen, JN, Jorgensen, BL, Grotinum, D & Gram, L 2004 The contribution of bacteriocin to inhibition of Listeria monocytogenes by Carnobacterium piscicola strains in cold-smoked salmon systems. Journal of Applied Microbiology 96 133–143CrossRefGoogle ScholarPubMed
Oscariz, JC & Pisabarro, AG 2001 Classification and mode of action of membrane-active bacteriocins produced by gram-positive bacteria. International Microbiology 4 13–19CrossRefGoogle ScholarPubMed
Pailin, T, Kang, DH, Schimidt, K & Fung, DYC 2001 Detection of extracellular bound proteinase in EPS-producing lactic acid bacteria cultures on skim milk agar. Letters in Applied Microbiology 33 45–49CrossRefGoogle ScholarPubMed
Rodriguez, E, Gonzalez, B, Gaya, P, Nunez, M & Medina, M 2000 Diversity of bacteriocins produced by lactic acid bacteria isolated from raw milk. International Dairy Journal 10 7–15CrossRefGoogle Scholar
Schnurer, J & Magnusson, J 2005 Antifungal lactic acid bacteria as biopreservatives. Trends in Food Science and Technology 16 70–78CrossRefGoogle Scholar
Settanni, L & Moschetti, G 2010 Non-starter lactic acid bacteria used to improve cheese quality and provide health benefits. Food Microbiology 27 691–697CrossRefGoogle ScholarPubMed
Sousa, MJ, Ardo, Y & McSweeney, PLH 2001 Advances in the study of proteolysis during cheese ripening. International Dairy Journal 11 327–345CrossRefGoogle Scholar
Stevens, KA, Sheldon, BW, Klapes, NA & Klaenhammer, TR 1991 Nisin treatment of Salmonella species and other gram-negative bacteria. Applied and Environmental Microbiology 57 3613–3615CrossRefGoogle ScholarPubMed
Torriani, S, Felis, GE & Dellaglio, F 2001 Differentiation of Lactobacillus plantarum, L. pentosus, and L. paraplantarum by recA gene sequence analysis and multiplex PCR assay with recA gene-derived primers. Applied and Environmental Microbiology 67 3450–3454CrossRefGoogle ScholarPubMed
Turner, S, Pryer, KM, Miao, VPW & Palmer, JD 1999 Investigating deep phylogenetic relationships among cyanobacteria and plastids by small submit rRNA sequence analysis. Journal of Eukaryotic Microbiology 46 327–338CrossRefGoogle Scholar
Voulgari, K, Hatzikamari, M, Delepoglou, A, Georgakopoulos, P, Litopoulou-Tzanetaki, E & Tzanetakis, N 2010 Antifungal activity of non-starter lactic acid bacteria isolates from dairy products. Food Control 21 136–142CrossRefGoogle Scholar
Ward, LJH & Timmins, MJ 1999 Differentiation of Lactobacillus casei, Lactobacillus paracasei and Lactobacillus rhamnosus by polymerase chain reaction. Letters in Applied Microbiology 29 90–92CrossRefGoogle ScholarPubMed