Presence of aprX gene in Pseudomonas spp. from refrigerated raw milk and their proteolytic ability
DOI:
https://doi.org/10.5433/1679-0359.2020v41n4p1421Palabras clave:
Psychrotrophic, Enzyme, Quality, Proteolysis.Resumen
This study aimed to determine the frequency of Pseudomonas fluorescens, P. putida, and P. aeruginosa in refrigerated raw milk; their proteolytic potential; and your association with the aprX gene. Of the 173 isolates confirmed as belonging to Pseudomonas spp., 37% were P. fluorescens, 25.4% P. putida, and none belongs to P. aeruginosa. The aprX gene was distributed proportionally between P. putida (68%) and P. fluorescens (75%), but it was not associated with low or high proteolytic potential in both species. P. putida (16) and P. fluorescens (14) isolates with no aprX gene identified also had proteolytic potential. Considering the synthesis of proteases other than AprX by the isolates under study, we concluded that P. fluorescens and P. putida represented 62.4% of the Pseudomonas genus, with high probability of having the aprX gene and proteolytic potential. However, there was no association between the deteriorating potential with the presence of the aprX gene.Métricas
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Almeida, K. M., Bruzaroski, S. R., Zanol, D., Melo, M., Santos, J. S., Aragon-Alegro, L. C., & Santana, E. H. W. (2017). Pseudomonas spp. and P. fluorescens: population in refrigerated raw milk. Ciência Rural, 47(1), 2-6. doi: 10.1590/0103-8478cr20151540
Bach, J. H., Hartmann, A., Schloter, M., & Munch, C. J. (2001). PCR primers and functional probes for amplification and detection of bacterial genes for extracellular peptidases in single strains and in soil. Journal of Microbiological Methods, 44(2), 173-182. doi: 10.1016/S0167-7012(00)00239-6
Decimo, M., Morandi, S., Machado, S. G., Bagliniere, F., & Vanetti, M. C. D. (2018). Psychrotrophic bacteria. In Poltronieri, P, Microbiology in dairy processing: challenges and opportunities (pp. 37-62). Pondicherry: Wiley Blackwell.
Decimo, M., Morandi, S., Silvetti, T., & Brasca, M. (2014). Characterization of Gram negative psychrotrophic bacteria isolated from Italian bulk tank milk. Journal of Food Science, 79(10), 2081-2090. doi: 10.1111/1750-3841.12645
Dogan, B., & Boor, K. J. (2003). Genetic diversity and spoilage potentials among Pseudomonas spp. isolated from fluid milk products and dairy processing plants. Applied and Environmental Microbiology, 69(1), 130-138. doi: 10.1128/AEM.69.1
Fagundes, C. M., Fischer, V., Silva, W. P., Carbonera, N., & Araujo, M. R. (2006). Presença de Pseudomonas spp. em função de diferentes etapas da ordenha com distintos manejos higiênicos e no leite refrigerado. Ciência Rural, Santa Maria, 36(2), 568-572. doi: 10.1590/S0103
Frank, J. F., & Yousef, A. E. (2004). Tests for groups of microorganisms. In M. W. Wehr, & J. F. Frank (Ed.), Standard methods for the examination of dairy products (pp. 227-247). New York: American Public Health Association.
Martins, M. L., Araújo, E. F., Mantovani, H. C., Moraes, C. A., & Vanetti, M. C. D. (2005). Detection of the apr gene in proteolytic psychrotrophic bacteria isolated from refrigerated raw milk. International Journal of Food Microbiology, 102(2), 203-211. doi: 10.1016/j.ijfoodmicro.2004.12.016
Meng, L., Zhang, Y., Liu, H., Zhao, S., Wang, J., & Zheng, N. (2017). Characterization of Pseudomonas spp. and associated proteolytic properties in raw milk stored at low temperatures. Frontiers of Microbiology, 8(8), 1-7. doi: 10.3389/fmicb.2017.02158 Confirmar o ano. No texto foi citado 2007.
Oliveira, G. B., Favarin, L., Luchese, R. H., & MClintosh, D. (2015). Psychrotrophic bacteria in milk: how much do we really know? Brazilian Journal of Microbiology, 46(2), 313-321. doi: 10.1590/S1517-838246220130963
Scarpellini, M., Franzetti, L., & Galli, A. (2004). Development of PCR assay to identify Pseudomonas fluorescens and its biotype. FEMS Microbiology Letters, 236(2), 257-260. doi: 10.1111/j.1574-6968.2004.tb09655.x
Spilker, T., Coenye, T., Vandame, P., & Lipuma J. J. (2004). PCR-Based assay for differentiation of Pseudomonas aeruginosa from other Pseudomonas species recovered from cystic fribrosis patients. Journal of Clinical Microbiology, 42(5), 2074-2079. doi: 10.1128/JCM.42.5.2074-2079
Stuknytë, M., Decimo, M., Colzani, M., Silvetti, T., Brasca, M., Cattaneo, S., Aldini, G., & De Noni, I. (2016). Extracellular thermostable proteolytic activity of the milk spoilage bacterium Pseudomonas fluorescens PS19 on bovine caseins. Journal of Dairy Science, 99(6), 4188-4195. doi: 10.3168/jds.2016-10894
Timmis, K. N. (2002). Pseudomonas putida: a cosmopolitan opportunist par excellence. Environmental Microbiology, 4(12), 779-781. doi: 10.1046/j.1462-2920.2002.00365.x
Xin, L., Zhaoxu, M., Zhang, L., Cui, Y., Han, X., & Yi, H. (2017). The diversity and proteolytic properties of psychrotrophic bacteria in raw cows’ milk from North China. International Dairy Journal, 66(1), 34-41, 2017. doi: 10.1016/j.idairyj.2016.10.014
Yamamoto, S., & Harayama, S. (1995). PCR amplification and direct sequencing of gyrB genes with universal primers and their application to the detection and taxonomic analysis of Pseudomonas putida strains. Applied and Environmental Microbiology, 61(3), 1104-1109.
Bach, J. H., Hartmann, A., Schloter, M., & Munch, C. J. (2001). PCR primers and functional probes for amplification and detection of bacterial genes for extracellular peptidases in single strains and in soil. Journal of Microbiological Methods, 44(2), 173-182. doi: 10.1016/S0167-7012(00)00239-6
Decimo, M., Morandi, S., Machado, S. G., Bagliniere, F., & Vanetti, M. C. D. (2018). Psychrotrophic bacteria. In Poltronieri, P, Microbiology in dairy processing: challenges and opportunities (pp. 37-62). Pondicherry: Wiley Blackwell.
Decimo, M., Morandi, S., Silvetti, T., & Brasca, M. (2014). Characterization of Gram negative psychrotrophic bacteria isolated from Italian bulk tank milk. Journal of Food Science, 79(10), 2081-2090. doi: 10.1111/1750-3841.12645
Dogan, B., & Boor, K. J. (2003). Genetic diversity and spoilage potentials among Pseudomonas spp. isolated from fluid milk products and dairy processing plants. Applied and Environmental Microbiology, 69(1), 130-138. doi: 10.1128/AEM.69.1
Fagundes, C. M., Fischer, V., Silva, W. P., Carbonera, N., & Araujo, M. R. (2006). Presença de Pseudomonas spp. em função de diferentes etapas da ordenha com distintos manejos higiênicos e no leite refrigerado. Ciência Rural, Santa Maria, 36(2), 568-572. doi: 10.1590/S0103
Frank, J. F., & Yousef, A. E. (2004). Tests for groups of microorganisms. In M. W. Wehr, & J. F. Frank (Ed.), Standard methods for the examination of dairy products (pp. 227-247). New York: American Public Health Association.
Martins, M. L., Araújo, E. F., Mantovani, H. C., Moraes, C. A., & Vanetti, M. C. D. (2005). Detection of the apr gene in proteolytic psychrotrophic bacteria isolated from refrigerated raw milk. International Journal of Food Microbiology, 102(2), 203-211. doi: 10.1016/j.ijfoodmicro.2004.12.016
Meng, L., Zhang, Y., Liu, H., Zhao, S., Wang, J., & Zheng, N. (2017). Characterization of Pseudomonas spp. and associated proteolytic properties in raw milk stored at low temperatures. Frontiers of Microbiology, 8(8), 1-7. doi: 10.3389/fmicb.2017.02158 Confirmar o ano. No texto foi citado 2007.
Oliveira, G. B., Favarin, L., Luchese, R. H., & MClintosh, D. (2015). Psychrotrophic bacteria in milk: how much do we really know? Brazilian Journal of Microbiology, 46(2), 313-321. doi: 10.1590/S1517-838246220130963
Scarpellini, M., Franzetti, L., & Galli, A. (2004). Development of PCR assay to identify Pseudomonas fluorescens and its biotype. FEMS Microbiology Letters, 236(2), 257-260. doi: 10.1111/j.1574-6968.2004.tb09655.x
Spilker, T., Coenye, T., Vandame, P., & Lipuma J. J. (2004). PCR-Based assay for differentiation of Pseudomonas aeruginosa from other Pseudomonas species recovered from cystic fribrosis patients. Journal of Clinical Microbiology, 42(5), 2074-2079. doi: 10.1128/JCM.42.5.2074-2079
Stuknytë, M., Decimo, M., Colzani, M., Silvetti, T., Brasca, M., Cattaneo, S., Aldini, G., & De Noni, I. (2016). Extracellular thermostable proteolytic activity of the milk spoilage bacterium Pseudomonas fluorescens PS19 on bovine caseins. Journal of Dairy Science, 99(6), 4188-4195. doi: 10.3168/jds.2016-10894
Timmis, K. N. (2002). Pseudomonas putida: a cosmopolitan opportunist par excellence. Environmental Microbiology, 4(12), 779-781. doi: 10.1046/j.1462-2920.2002.00365.x
Xin, L., Zhaoxu, M., Zhang, L., Cui, Y., Han, X., & Yi, H. (2017). The diversity and proteolytic properties of psychrotrophic bacteria in raw cows’ milk from North China. International Dairy Journal, 66(1), 34-41, 2017. doi: 10.1016/j.idairyj.2016.10.014
Yamamoto, S., & Harayama, S. (1995). PCR amplification and direct sequencing of gyrB genes with universal primers and their application to the detection and taxonomic analysis of Pseudomonas putida strains. Applied and Environmental Microbiology, 61(3), 1104-1109.
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2020-05-13
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Longhi, R. D., Bruzaroski, S. R., Eleodoro, J. I., Poli-Frederico, R. C., Fagnani, R., Ludovico, A., & Santana, E. H. W. de. (2020). Presence of aprX gene in Pseudomonas spp. from refrigerated raw milk and their proteolytic ability. Semina: Ciências Agrárias, 41(4), 1421–1426. https://doi.org/10.5433/1679-0359.2020v41n4p1421
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