Previously we described a rapid fluorogenic method to measure the activity of five bacteriocins produced by Mexican strains of Bacillus thuringiensis against B. cereus 183. Here we standardize this method to efficiently determine the activity of bacteriocins against both Gram-positive and Gram-negative bacteria. It was determined that the crucial parameter required to obtain reproducible results was the number of cells used in the assay, that is, ~4 × 108 cell/mL and ~7 × 108 cell/mL, respectively, for target Gram-positive and Gram-negative bacteria. Comparative analyses of the fluorogenic and traditional well-diffusion assays showed correlation coefficients of 0.88 to 0.99 and 0.83 to 0.99, respectively, for Gram-positive and Gram-negative bacteria. The fluorogenic method demonstrated that the five bacteriocins of B. thuringiensis have bacteriolytic and bacteriostatic activities against all microorganisms tested, including clinically significant bacteria such as Listeria monocytogenes, Proteus vulgaris, and Shigella flexneri reported previously to be resistant to the antimicrobials as determined using the well-diffusion protocol. These results demonstrate that the fluorogenic assay is a more sensitive, reliable, and rapid method when compared with the well-diffusion method and can easily be adapted in screening protocols for bacteriocin production by other microorganisms.
Las quitinasas sintetizadas por plantas, hongos, insectos y bacterias tienen un gran potencial debido a su amplio espectro de aplicaciones. En este trabajo revisamos generalidades sobre las quitinasas y quitosanasas bacterianas y su uso en la producción de quito-oligosacáridos. Este tipo de biomoléculas han creado un mercado biotecnológico diversificado e ilimitado que incluye aplicaciones en alimentos como aditivos y bioconservadores, asimismo en múltiples aplicaciones biomédicas enfocadas a la actividad anti-tumoral, a la capacidad como agentes antioxidantes y como antidiabéticos. Además, en la agricultura se han aplicado como factores de nodulación, como agentes osmoprotectores y antioxidantes para beneficiar el crecimiento de cultivos. Es importante destacar el potencial de las quitinasas sintetizadas por Bacillus thuringiensis, el bioinsecticida mas importante mundialmente. Las quitinasas de B. thuringiensis se han empleado recientemente para generar quito-oligosacáridos que tienen actividad antimicrobiana, particularmente contra diversas bacterias patógenas de importancia en salud pública transmitidas por alimentos.
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