Con la finalidad de obtener bacterias ácido lácticas productoras de bacteriocinas se recolectaron 33 muestras de quesos artesanales provenientes de Lima y provincias y se aislaron 341 cepas de lactobacilos según los procedimientos establecidos por el Manual de Bergey. Las especies de lactobacilos aisladas con mayor frecuencia fueron Lactobacillus casei 56 % (191) y Lactobacillus plantarum 35.8 % (122). Sólo el 16.42 % (56/341) de los aislados presentaron actividad bacteriocinogénica frente a Lactobacillus acidophilus ATCC 4356. Los sobrenadantes de los cultivos bacteriocinogénicos se ajustaron el pH a 6.5 para descartar la acción de ácidos orgánicos, además se determinó el péroxido de hidrógeno por acción de la catalasa. La constitución bioquímica de las bacteriocinas presentes en los sobrenadantes fue determinada enzimáticamente utilizando pronasa E, a -amilasa y lipasa. El 53 % (30) de las bacteriocinas fueron de naturaleza lipoproteica, 25 % (14) glicolipoproteica, 13 % (7) glicoproteica y 9 % (5) solamente proteica. Por otro lado, los sobrenadantes fueron sometidos a tratamientos térmicos de 60, 80 y 100 °C, a pH 4, 7 y 9, y a temperaturas de conservación de 4, 15 y 32 °C y se encontró que la termoestabilidad fue de 60 a 80 °C, el pH óptimo de 4 a 7 y la temperatura óptima de conservación de 4 a 32 °C. Las bacteriocinas presentes en los sobrenadantes de los diferentes aislados inhibieron el crecimiento de cepas de Bacillus cereus UA05, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 10536, Salmonella typhimurium ATCC 14028, Salmonella choleraesuis ATCC 10708, Shigella sonnei ATCC 9290 y Pseudomonas aeruginosa ATCC 9027. Sin embargo, Listeria innocua UP01 fue resistente. El aislado identificado como Lactobacillus plantarum LM1 produjo la bacteriocina con los mejores parámetros fisicoquímicos experimentados y además presentó un amplio espectro de inhibición contra los microorganismos indicadores evaluados.
Palabras Clave: bacterias ácido lácticas, bacteriocinas, lactobacilos, Lactobacillus plantarum, quesos artesanales / The aim of this study was to get bacteriocin – producing lactic acid bacteria. For the purpose of this study, thirty three samples of artisanal cheeses from Lima and provinieses were collected, and three hundred and forty one strains of lactobacilli were isolated according to the procedures established by Bergey’s Manual. The most frequently isolated species were Lactobacillus casei 56% (191) and Lactobacillus plantarum 35.8% (122).
Only 16.42% (56/341) of the isolates showed bacteriocinogenic activity against Lactobacillus acidophilus ATCC 4356. The supernatants of bacteriocinogenic cultures were adjusted to pH 6.5 to rule out the action of organic acids, and it was determined the presence of hydrogen peroxide by the assay with catalase. The biochemical constitution of the bacteriocins present in the supernatants was determined using enzymatically pronase E, a - amylase and lipase. The 53% (30) of bacteriocins was of lipoproteinaceous nature, 25% (14) of glycolipoproteinaceous nature, 13% (7) of glycoproteinaceous nature, and 9% (5) only of proteinaceous nature. On the other hand, the supernatants were exposed to various heat treatments at 60, 80, and 100°C, to pH variation at 4, 7, and 9, and to storage temperatures at 4, 15 and 32°C, and it was found that the thermostability of bacteriocins was found from 60 to 80°C, the optimum pH was from 4 to 7, and the optimum storage temperature was from 4 to 32°C. The bacteriocins present in the supernatants of the different isolates inhibited the growth of Bacillus cereus UA05, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 10536, Salmonella typhimurium ATCC 14028, Salmonella choleraesuis ATCC 10708, Shigella sonnei ATCC 9290 and Pseudomonas aeruginosa ATCC 9027. However, Listeria innocua UP01was resistant. The isolate identified as Lactobacillus plantarum LM1 produced the bacteriocin with the best physicochemical parameters, and showed a broad spectrum of inhibition against the indicator microorganisms.
Key words: Lactic acid bacteria, bacteriocins, lactobacilli, Lactobacillus plantarum, artisanal cheeses. / Tesis
| Identifer | oai:union.ndltd.org:Cybertesis/oai:cybertesis.unmsm.edu.pe:cybertesis/235 |
| Date | January 2008 |
| Creators | Cristóbal Delgado, Ruth Liliana, Cristóbal Delgado, Ruth Liliana |
| Publisher | Universidad Nacional Mayor de San Marcos |
| Source Sets | Universidad Nacional Mayor de San Marcos - SISBIB PERU |
| Language | Spanish |
| Detected Language | English |
| Type | info:eu-repo/semantics/bachelorThesis |
| Format | application/pdf |
| Source | Universidad Nacional Mayor de San Marcos, Repositorio de Tesis - UNMSM |
| Rights | info:eu-repo/semantics/openAccess |
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