EVALUACIÓN DE LA ACTIVIDAD CICLOHEXANONA MONOOXIGENASA EN CEPAS DE HONGOS FILAMENTOSOS Y LEVADURAS
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Rev. Bol. Quim
Abstract
Ciclohexanona monooxigenasa (CHMO) cataliza la oxigenación de cetonas lineales o cetonas cíclicas, produciendo así sus correspondientes lactonas, estas lactonas a nivel industrial son obtenidas por síntesis química. En cambio, la biotransformación tiene la ventaja de ser altamente enantio y regio selectivas dando productos puros de alto valor, además de contribuir al desarrollo de procesos amigables con el medio ambiente. Se evaluó la actividad CHMO de 90 cepas fúngicas las cuales fueron divididas en dos grupos: 85 cepas de hongos filamentosos que fueron obtenidas del cepario del Instituto de Investigaciones Fármaco Bioquímicas (IIFB) y 5 cepas de levaduras aisladas de tierras contaminadas con petróleo. Tres métodos de screening para CHMO fueron empleados: (i) por medio de la evaluación cualitativa por cambio de pH (viraje de color del indicador), se identificaron 3 cepas con potencial actividad enzimática, de las cuales se observó que la cepa 177 QD presentó un consumo de 41 mM al lapso de 30 horas, (ii) mediante ensayo colorimétrico para determinación de cetonas cíclicas; posteriormente se midió la actividad enzimática (iii) mediante la monitorización espectrofotométrica de la depleción de nicotinamida adenina dinucleótido fosfato (NADPH), la cual dio una actividad de 90 UI/L. Se procedió a identificar la cepa más activa mediante sus características macroscópicas y microscópicas, que la relacionan con Aspergillus flavas
Cyclohexanone monooxygenase (CHMO) catalyzes the oxygenation of linear or cyclic ketone ketone, thereby producing the corresponding lactones, these lactones on an industrial level are obtained by chemical synthesis. Instead, the biotransformation has the advantage of being highly enantioselective and regio-selective giving puré producís of high valué, and contribute to the development of processes environmentally friendly. CHMO activity was assessed 90 fungal strains which were divided into two groups: 85 strains of filamentous fungi were obtained from the culture collection of Biochemical Drug Research Institute (IIFB) and 5 strains of yeasts isolated from soil contaminated with oil. Three methods were employed CHMO screening for: (i) by means of the qualitative evaluation by pH change (color change indicator) identified three strains with potential enzymatic activity, which was observed for the strain introduced QD 177 a consumption of 41 mM to 30 hour period, by (ii) colorimetric assay for determination of cyclic ketones, and subsequently the enzyme activity was measured by the (iii) spectrophotometric monitoring depletion of nicotinamide adenine dinucleotide phosphate (NADPH), which gave an activity of 90 IU / L. We proceeded to identify the most active strain by macroscopic and microscopic characteristics, which relate to Aspergillus flavas.
Cyclohexanone monooxygenase (CHMO) catalyzes the oxygenation of linear or cyclic ketone ketone, thereby producing the corresponding lactones, these lactones on an industrial level are obtained by chemical synthesis. Instead, the biotransformation has the advantage of being highly enantioselective and regio-selective giving puré producís of high valué, and contribute to the development of processes environmentally friendly. CHMO activity was assessed 90 fungal strains which were divided into two groups: 85 strains of filamentous fungi were obtained from the culture collection of Biochemical Drug Research Institute (IIFB) and 5 strains of yeasts isolated from soil contaminated with oil. Three methods were employed CHMO screening for: (i) by means of the qualitative evaluation by pH change (color change indicator) identified three strains with potential enzymatic activity, which was observed for the strain introduced QD 177 a consumption of 41 mM to 30 hour period, by (ii) colorimetric assay for determination of cyclic ketones, and subsequently the enzyme activity was measured by the (iii) spectrophotometric monitoring depletion of nicotinamide adenine dinucleotide phosphate (NADPH), which gave an activity of 90 IU / L. We proceeded to identify the most active strain by macroscopic and microscopic characteristics, which relate to Aspergillus flavas.
Description
Vol. 30, No. 1