A catecholase from Laccaria laccata a wild edible mushroom and its catalytic efficiency in organic media

KOLCUOĞLU Y., KUYUMCU I., ÇOLAK A.

JOURNAL OF FOOD BIOCHEMISTRY, cilt.42, sa.5, 2018 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 42 Sayı: 5
  • Basım Tarihi: 2018
  • Doi Numarası: 10.1111/jfbc.12605
  • Dergi Adı: JOURNAL OF FOOD BIOCHEMISTRY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Anahtar Kelimeler: affinity chromatography, catecholase, diphenolase, Laccaria laccata, pH-stability, thermal stability, POLYPHENOL OXIDASE, DIPHENOLASE ACTIVITIES, THERMAL INACTIVATION, AGARICUS-BISPORUS, SOLVENT MEDIA, PURIFICATION, TYROSINASE, L., BIOCATALYSIS, CHROMATOGRAPHY
  • Karadeniz Teknik Üniversitesi Adresli: Evet

Özet

In this study, catecholase was purified from Laccaria laccata by affinity chromatography and the enzyme activity was investigated in organic media. Among the tested substrates, the highest catecholase activity was observed in 4-MC, DHPPA, and L-DOPA. A single band around 58.1kDa was observed on SDS-PAGE of the purified enzyme. Km values were calculated as 0.25, 0.40, and 0.83mM for 4-MC, DHPPA, and L-DOPA, respectively. The highest V-max value was calculated as 2500 U/mg protein for 4-MC. The inhibitors used in this study showed mixed type inhibition kinetics for all tested substrates. Ascorbic acid was found to be most effective inhibitor with low K-i value. Also, it was observed that LacPPO is more stable at optimum pH and temperature. In addition, LacPPO was found to be an effective biocatalyst in organic medium. According to the results obtained, the purified enzyme may be useful for some industrial and/or clinical purposes.