PROTEOLYTIC POTENTIAL OF PSEUDOMONAS FLUORESCENS ISOLATED FROM REFRIGERATED RAW MILK

Authors

  • Cláudia Lúcia Oliveira Pinto1 EPAMIG
  • Solimar Gonçalves Machado Federal University of Viçosa, Viçosa
  • Rodrigo Rezende Cardoso Federal University of Viçosa
  • Rita Maria Alves Brazilian Agricultural Research Corporation's, Embrapa Wheat, Passo Fundo, RS, Brazil
  • Maria Cristina Dantas Vanetti Federal University of Viçosa, Viçosa

DOI:

https://doi.org/10.21206/rbas.v4i2.254

Abstract

The growth rate and the proteolytic activity of Pseudomonas fluorescens strains 07A and 041, isolated from cow’s milk, were evaluated at 2, 4, 7 and 10ºC. P. fluorescens promoted protein degradation during storage of milk samples as observed by Proteolytic activity measurement, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and heat stability of milk. Casein hydrolysis resulted in loss of thermal stability of milk and in formation of fragments of low and medium molecular mass. Temperatures up to 10°C did not
guarantee raw milk quality when contamination by P. fluorescens was equal or higher than 106 cfu/mL.

Downloads

Download data is not yet available.

Author Biographies

Cláudia Lúcia Oliveira Pinto1, EPAMIG

Agriculture and Livestock Research Corporation's of Minas Gerais, EPAMIG, Viçosa, MG, Brasil. 

Solimar Gonçalves Machado, Federal University of Viçosa, Viçosa

Department of Microbiology, Federal University of Viçosa, Viçosa, 36570-000, MG, Brazil.

Rodrigo Rezende Cardoso, Federal University of Viçosa

Department of Food Science and Technology, Federal University of Viçosa, Viçosa, 36570-000, MG, Brazil.

Rita Maria Alves, Brazilian Agricultural Research Corporation's, Embrapa Wheat, Passo Fundo, RS, Brazil

Brazilian Agricultural Research Corporation's, Embrapa Wheat, Passo Fundo, RS, Brazil

Maria Cristina Dantas Vanetti, Federal University of Viçosa, Viçosa

Department of Microbiology, Federal University of Viçosa, Viçosa, 36570-000, MG, Brazil.

References

ADAMS, D.M.; BARACH, J.T.; SPECK, M.L. Effect of psychrotrophic bacteria from raw milk on milk proteins and stability of milk proteins to
ultrahigh temperature treatment. Journal of Dairy Science, v.59, n.5, p.823-827, 1976.

BAGLINIÈRE, F.; TANGUY, G.; JARDIN, J. et al. Quantitative and qualitative variability of the caseinolytic potential of different strains of
Pseudomonas fluorescens: implications for the stability of casein micelles of UHT milks during their storage. Food Chemistry, v.135, n.4, p.2593-2603, 2012.

BRASIL. Regulamento Técnico de Produção, Identidade e Qualidade do Leite tipo A, Regulamento Técnico de Identidade e Qualidade de Leite Cru Refrigerado, Regulamento Técnico de Identidade e Qualidade de Leite Pasteurizado e Regulamento Técnico da Coleta de Leite Cru Refrigerado e seu Transporte a Granel. Instrução Normativa nº 62, de 29 de de dezembro de 2011.

Diário Oficial [da] República Federativa do Brasil, Brasília, DF: Ministério da Agricultura, Pecuária e Abastecimento: p.6-37, 2011.

BRAUN, P.; SUTHERLAND, J.P. Predictive modelling of growth and enzyme production and activity by a cocktail of Pseudomonas spp.,
Shewanella putrefaciens and Acinetobacter. International Journal of Food
Microbiology, v.86, n.3, p.271-282, 2003.

BROCK, T.D. Biology of microorganisms. 7th Ed. New Jersey: Prentice Hall, 1994, 909p.

CELESTINO, E.L.; IYER, M.; ROGINSKI, H. Reconstituted UHT-treated milk: Effects of raw milk, powder quality and storage conditions of
UHT milk on its physico-chemical attributes and flavour. International Dairy Journal, v.7, n.2-3, p.129-140, 1997.

CHEN, L.; DANIEL, R.M.; COOLBEAR, T. Detection and impact of protease and lipase activities in milk and milk powders. International Dairy Journal, v.13, n.4, p.255-275, 2003.

COSTA, L.M.; GÓMEZ, F.S.; MOLINA, L.H.C. et al. Purificación y caracterización de proteasas de Pseudomonas fluorescens y sus efectos sobre las proteínas de la leche. Archivos Latinoamericanos de Nutrición, v.52, n.2, p.1-13, 2002.

CRUDDEN, A.; FOX, P.F.; KELLY, A.L. Factors affecting the hydrolytic action of plasmin in milk. International Dairy Journal, v.15, n.4,
p.305-313, 2005.

DECIMO, M.; MORANDI, S.; SALVETTI, T. et al. Characterization of gram-negative psychrotrophic bacteria isolated from Italian bulk tank milk. Journal of Food Science, v.79, n.10, p.M2081-M2090, 2014.

DE JONGHE, V.; COOREVITS, A.; VAN HOORDE, K. et al. Influence of storage conditions on the growth of Pseudomonas species in refrigerated
raw milk. Applied and Environmental Microbiology, v.77, n.2, p.460-470, 2011.

DOGAN, B.; BOOR, K.J. Genetic diversity and spoilage potentials among Pseudomonas spp. isolated from fluid milk products and dairy
processing plants. Applied and Environmental Microbiology, v.69, n.1,
p.130-138, 2003.

ENEROTH, A.; AHRNÉ, S.; MOLIN, G. Contamination routes of Gram-negative spoilage bacteria in the production of pasteurized milk,
evaluated by randomly amplified polymorphic DNA (RAPD). International Dairy Journal, v.10, n.5-6, p.325-331, 2000.

FAIRBAIRN, D.J.; LAW, B.A. Proteinase of psychrotrophic bacteria: their production, properties, effects and control. Journal of Dairy Research, v.53, n.1, p.139-77, 1986.

FRANK, J.F.; CHRISTEN, G.L.; BULLERMAN, L.B. Tests for groups of microorganisms. In: MARSHALL, R.T. (Ed.). Standard methods
for the examination of dairy products. New York: APHA, p.271-286, 1992.

HACHANA, Y.; KRAIEM, K.; PAAPE, M.J. Effect of plasmin, milk somatic cells and psychrotrophic bacteria on casein fractions of ultra high temperature treated milk. Food Science and Technology, v.16, n.1, p.79-
86, 2010.

HORNE, D.S. Casein micelle structure: Models and muddles. Current Opinion in Colloid & amp; Interface Science, v.11, n.2-3, p.148-153, 2006.

HULL, M.E. Studies on milk proteins. II Colorimetric determination of the partial hydrolysis of the proteins in milk. Journal of Dairy Science, v.30, p.881-894, 1947.

KUMARESAN, G.; ANNALVILLI, R.; SIVAKUMAR, K. Psychrotrophic spoilage of raw milk at different temperatures of storage. Journal of Applied Sciences Research, v.3, n.11, p.1383-1387, 2007.

LAEMMLI, U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, v.227, n.5259, p.680-685, 1970.

LARSON, N.K.; ISMAIL, B., NIELSEN, S.S. et al. Activity of Bacillus polymyxa protease on components of the plasmin system in milk.
International Dairy Journal, v.16, n.6, p.586-592, 2006.

MARCHAND, S.; HEYLEN, K.; MESSENS, W. et al. Seasonal influence on heat-resistant proteolytic capacity of Pseudomonas lundensis and Pseudomonas fragi, predominant milk spoilers isolated from Belgian raw milk samples. Environmental Microbiology, v.11, n.2, p.467-482, 2009a.

MARCHAND, S.; VANDRIESCHE, G.; COOREVITS, A. et al. Heterogeneity of heatresistant proteases from milk Pseudomonas species. International Journal of Food Microbiology, v.133, n.1-2, p.68-77, 2009b.

MARTINS, M.L.; ARAÚJO, E.F.; MANTOVANI, H.C. et al. Detection of the apr gene in proteolytic psychrotrophic bacteria isolated from refrigerated raw milk. International Journal of Food Microbiology, v.102, n.2, p.203-
211, 2005.

MARTINS, M.L.; PINTO, C.L.O.; ROCHA, R.B. et al. Genetic diversity of Gram-negative, proteolytic, psychrotrophic bacteria isolated from
refrigerated raw milk. International Journal of Food Microbiology, v.111, n.2, p.144-148, 2006.

MORTON, R.D. Aerobic plate count. In:DOWNES, F.P. and ITO, K. (Ed.).
Compendium of Methods for the Microbiological Examination of
Foods. 4th Ed. Washington: American Public Health Association, p.63-67, 2001.

MUNSCH-ALATOSSAVA, P.; ALATOSSAVA, T. Phenotypic characterization of raw milkassociated psychrotrophic bacteria. Microbiological Research, v.161, n.4, p.334-346, 2006.

NERO, L.A.; VIÇOSA, G.N.; PEREIRA, F.E.V. Qualidade Microbiológica do leite determinada por características de produção. Ciência e Tecnologia de Alimentos, v.29, n.2, p.386-390, 2009.

NICODÈME, M.; GRILL, J.-P.; HUMBERT, G. et al. Extracellular protease activity of different Pseudomonas strains: dependence of proteolytic
activity on culture conditions. Journal of Applied Microbiology, v.99, p.641-648, 2005.

NIELSEN, S.S. Plasmin system and microbial proteases in milk: characteristics, roles and relationship. Journal of Agricultural and
Food Chemistry, v.50, n.22, p.6628-6624, 2002.

PINTO, C.L.O.; MARTINS, M.L.; VANETTI, M.C.D. Qualidade microbiológica de leite cru refrigerado e isolamento de bactérias
psicrotróficas proteolíticas. Ciência e Tecnologia de Alimentos, v.26, n.3, p.645-651, 2006.

RAATS, D.; OFFEK, M.; MINZ, D. et al. Molecular analysis of bacterial communities in raw cow milk and the impact of refrigeration on its structure and dynamics. Food Microbiology, v.28, n.3, p.465-471, 2011.
R CORE TEAM. R: A language and environment for statistical computing. 2013. In: http://www.Rproject.org/ (accessed on January 4th 2015).

SØRHAUG, T.; STEPANIAK, L. Psychrotrophs and their enzymes in milk and dairy products: Quality aspects. Trends in Food Science & Technology, v.8, n.2, p.35-41, 1997.

TEH, K.H.; FLINT, S.; Palmer, J. et al. Proteolysis produced within biofilms of bacterial isolates from raw milk tankers. International
Journal of Food Microbiology, v.157, n.1, p.28-34, 2012.

Published

2014-12-30

How to Cite

Pinto1, C. L. O., Machado, S. G., Cardoso, R. R., Alves, R. M., & Vanetti, M. C. D. (2014). PROTEOLYTIC POTENTIAL OF PSEUDOMONAS FLUORESCENS ISOLATED FROM REFRIGERATED RAW MILK. Brazilian Journal of Sustainable Agriculture, 4(2). https://doi.org/10.21206/rbas.v4i2.254

Most read articles by the same author(s)