Identificación de compuestos antimicrobianos tipo bacteriocina producidos por aislados de Pseudomas sp., análisis de su expresión In-Vitro y aplicación en vehículos de dispersión
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Fecha
2013
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Facultad de Ciencias Biológicas
Escuela de Ingeniería en Biotecnología
Three isolates of Pseudomonas sp. with anti-bacterial activity obtained from Antarctic territory were studied, from which a highly active bacteriocin Iike substance compound was found in 6-Al isolate, with activity against Escherichía colí TOP1O and Staphylococcus aureus 25923. The efficiency was registered as probably higher than many batenocins and higher than antibiotics available on the market. Bacteriocin like compound purification was perfonned using solid phase extraction and high performance liquid chromatography (HPLC). The substance was identified as CEMG-1, with a molecular mass lower than 21 kDa, pmtelnase K sensithre, themo-stabk at 80° and lethal for botñ studled stralns. Finally, its activity and eventual interferences were assayed with commonly used food additives in meat industry; saline solution, polyethylenglycol, 1% alginate and 50% glycerol. A reduction of its activity was elucidated in addition with saline solution (16X) and polyethylengycol (10%). In combination with alginate, the activity was the same. Even though its bacteriocin nature was not confimed completely, according to the resuits, it classifes in BLIS category with high activity; 0,12 pg/mL M.I.C. for E. coli TOP1O y 0,056 pglmL for S. aureus 25923. Additionaliy, its high bactericidal activity against Gram positive and Gram negative bacteria is interesting for preservative addiüve use in meat food industry.
Escuela de Ingeniería en Biotecnología
Three isolates of Pseudomonas sp. with anti-bacterial activity obtained from Antarctic territory were studied, from which a highly active bacteriocin Iike substance compound was found in 6-Al isolate, with activity against Escherichía colí TOP1O and Staphylococcus aureus 25923. The efficiency was registered as probably higher than many batenocins and higher than antibiotics available on the market. Bacteriocin like compound purification was perfonned using solid phase extraction and high performance liquid chromatography (HPLC). The substance was identified as CEMG-1, with a molecular mass lower than 21 kDa, pmtelnase K sensithre, themo-stabk at 80° and lethal for botñ studled stralns. Finally, its activity and eventual interferences were assayed with commonly used food additives in meat industry; saline solution, polyethylenglycol, 1% alginate and 50% glycerol. A reduction of its activity was elucidated in addition with saline solution (16X) and polyethylengycol (10%). In combination with alginate, the activity was the same. Even though its bacteriocin nature was not confimed completely, according to the resuits, it classifes in BLIS category with high activity; 0,12 pg/mL M.I.C. for E. coli TOP1O y 0,056 pglmL for S. aureus 25923. Additionaliy, its high bactericidal activity against Gram positive and Gram negative bacteria is interesting for preservative addiüve use in meat food industry.
Idioma
es
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Universidad Andrés Bello
Nombre de Curso
Licencia CC
Licencia CC
Resumen
Se estudiaron tres aislados de Pseudomonas sp. con actividad antibacteriana obtenidas desde
territorio antártico, de las cuales se logró identificar un compuesto tipo bacteriocina producido por el aislado 6-Al, con actividad antibacteriana contra Staphylococcus aureus 25923 y Escherichia coli TOP1O, posiblemente más eficiente que las bacteriocinas disponibles en el mercado y más eficiente que varios antibióticos. El compuesto fue purificado mediante extracción en fase sólida y
cromatografía líquida de alta resolución (HPLC), tras lo cual se identificó como CEMG-1, con una masa molecular inferior a los 21 kDa, sensible a proteinasa K, termoestable a 80° y bactericida para ambos casos estudiados.
Finalmente, se ensayó su actividad en conjunto a aditivos usados en la industria alimentaria de
la carne como; solución salina, polietilenglicol, alginato 1% y glicerol 50%, para evaluar sí su actividad se ve afectada por estos compuestos. Se evidenció una disminución de actividad en solución salina (16%) y polietilenglicol (10%). En el caso del alginato, la actividad se mantuvo al mismo nivel.
Aunque no se pudo confirmar su naturaleza, según los resultados obtenidos clasifica dentro
de sustancias inhibitorias similares a bacteriocina (BLIS) y con alta actividad; 0,12 pg/mL C.M.I. para E. colí TOP1O y 0,056 pg/mL para S. aureus 25923. La proyección que podría tener en la industria de la
carne para inhibir patógenos Gram positivo y Gram negaivo en condiciones adversas, resulta muy interesante.
Notas
Tesis (Magíster en Biotecnología)
Palabras clave
Pseudomonas, Aislamiento y Purificación, Bacteriocinas