Skip to main content Accessibility help

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

  • Fabricio L Tulini (a1), Nolwenn Hymery (a2), Thomas Haertlé (a3), Gwenaelle Le Blay (a4) and Elaine C P De Martinis (a1)...


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.


Corresponding author

*For correspondence; e-mail:


Hide All
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 131140
Axelsson, LT 1993 Lactic acid bacteria: classification and physiology. In Lactic Acid Bacteria, pp. 163 (Eds Salminenand, S & Wright, AT). New York: Marcel Dekker
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 842848
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 97106
Chanos, P & Williams, DR 2011 Anti-Listeria bacteriocin-producing bacteria from raw ewe's milk in northern Greece. Journal of Applied Microbiology 110 757768
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 93109
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 12
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 185190
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 206213
De Man, JD, Rogosa, M & Sharpe, ME 1960 A medium for the cultivation of Lactobacilli. Journal of Applied Bacteriology 23 130135
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 576587
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 2427
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 758764
Food and Drug Administration 2012 Summary of data concerning the safety and GRAS determination of food ferment solutions for use as a food ingredient. (24 February 2015)
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 12
Galvez, A, Abriouel, H, Lopez, RL & Ben Omar, N 2007 Bacteriocin-based strategies for food biopreservation. International Journal of Food Microbiology 120 5170
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 13161325
Hartemink, R, Van Laere, KM & Rombouts, FM 1997 Growth of enterobacteria on fructo-oligosaccharides. Journal of Applied Microbiology 83 367374
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 49304933
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 35583565
Lane, DJ 2001 16S/23S rRNA sequencing. In Nucleic Acid Techniques in Bacterial Systematics, pp. 115175 (Eds Stackebrandt, E & Goodfellow, M). New York, USA: John Wiley & Sons
Lee, HJ, Park, SY & Kim, J 2000 Multiplex PCR-based detection and identification of Leuconostoc species. FEMS Microbiology Letters 193 243247
Lee, KW, Park, JY, Jeong, HR, Heo, HJ, Han, NS & Kim, JH 2012 Probiotic properties of Weissella strains isolated from human faeces. Anaerobe 18 96102
Leigh, JA 1999 Streptococcus uberis: a permanent barrier to the control of bovine mastitis? Veterinary Journal 157 225238
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 16831688
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 261270
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 133143
Oscariz, JC & Pisabarro, AG 2001 Classification and mode of action of membrane-active bacteriocins produced by gram-positive bacteria. International Microbiology 4 1319
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 4549
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 10 715
Schnurer, J & Magnusson, J 2005 Antifungal lactic acid bacteria as biopreservatives. Trends in Food Science and Technology 16 7078
Settanni, L & Moschetti, G 2010 Non-starter lactic acid bacteria used to improve cheese quality and provide health benefits. Food Microbiology 27 691697
Sousa, MJ, Ardo, Y & McSweeney, PLH 2001 Advances in the study of proteolysis during cheese ripening. International Dairy Journal 11 327345
Stevens, KA, Sheldon, BW, Klapes, NA & Klaenhammer, TR 1991 Nisin treatment of Salmonella species and other gram-negative bacteria. Applied and Environmental Microbiology 57 36133615
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 34503454
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 327338
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 136142
Ward, LJH & Timmins, MJ 1999 Differentiation of Lactobacillus casei, Lactobacillus paracasei and Lactobacillus rhamnosus by polymerase chain reaction. Letters in Applied Microbiology 29 9092


Related content

Powered by UNSILO

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

  • Fabricio L Tulini (a1), Nolwenn Hymery (a2), Thomas Haertlé (a3), Gwenaelle Le Blay (a4) and Elaine C P De Martinis (a1)...


Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.