Purification and comparative characterization of monophenolase and diphenolase activities from a wild edible mushroom (Macrolepiota gracilenta)


Kolcuoglu Y.

PROCESS BIOCHEMISTRY, cilt.47, ss.2449-2454, 2012 (SCI İndekslerine Giren Dergi) identifier identifier

  • Cilt numarası: 47 Konu: 12
  • Basım Tarihi: 2012
  • Doi Numarası: 10.1016/j.procbio.2012.10.008
  • Dergi Adı: PROCESS BIOCHEMISTRY
  • Sayfa Sayıları: ss.2449-2454

Özet

Polyphenol oxidases (PPO) are very important enzymes group in many industrial applications, especially in food, medicine and cosmetics. PPO from Macrolepiota gracilenta, a wild edible mushroom, was purified using a Sepharose 4B-L-tyrosine-p-amino benzoic acid affinity column and characterized in terms of mono- and diphenolase activity. The highest activities for pure enzyme were observed in the presence of PHPPA and DHPPA for monophenolase and diphenolase, respectively. The enzyme showed pH optimum values at 7.0 and 5.0, respectively, for monophenolase and diphenolase activities. K-m values calculated as 0.8 mM for monophenolase and I mM for diphenolase activity at the presence of PHPPA and DHPPA as substrate, respectively. V-max values were calculated as 2000 U/mg protein for both activity. Monophenolase and diphenolase activities were conserved approximately 40% and 60%, respectively, in their optimum pH at 4 degrees C after 5 day incubation. The activities were inhibited most effectively by thiourea. The data obtained from this study showed that this enzyme could be useful for some industrial purposes. (C) 2012 Elsevier Ltd. All rights reserved.

Polyphenol oxidases (PPO) are very important enzymes group in many industrial applications, especially in food, medicine and cosmetics. PPO from Macrolepiota gracilenta, a wild edible mushroom, was purified using a Sepharose 4B-L-tyrosine-p-amino benzoic acid affinity column and characterized in terms of mono- and diphenolase activity. The highest activities for pure enzyme were observed in the presence of PHPPA and DHPPA for monophenolase and diphenolase, respectively. The enzyme showed pH optimum values at 7.0 and 5.0, respectively, for monophenolase and diphenolase activities. K-m values calculated as 0.8 mM for monophenolase and I mM for diphenolase activity at the presence of PHPPA and DHPPA as substrate, respectively. V-max values were calculated as 2000 U/mg protein for both activity. Monophenolase and diphenolase activities were conserved approximately 40% and 60%, respectively, in their optimum pH at 4 degrees C after 5 day incubation. The activities were inhibited most effectively by thiourea. The data obtained from this study showed that this enzyme could be useful for some industrial purposes.