Production, purification, and characterization of a thermo-alkali stable and metal-tolerant carboxymethylcellulase from newly isolated Bacillus methylotrophicus Y37


Duman Y. , Yüzügüllü Karakuş Y. , Sertel A., Polat F.

TURKISH JOURNAL OF CHEMISTRY, cilt.40, ss.802-815, 2016 (SCI İndekslerine Giren Dergi)

  • Cilt numarası: 40
  • Basım Tarihi: 2016
  • Doi Numarası: 10.3906/kim-1602-55
  • Dergi Adı: TURKISH JOURNAL OF CHEMISTRY
  • Sayfa Sayısı: ss.802-815

Özet

A carboxymethylcellulose (CMC)-degrading bacterium was isolated from soil, identified as Bacillus methylotrophicus according to the physiological properties and analyses of 16S rRNA and a partial sequence of the gyrase A (gyrA) gene, and named as B. methylotrophicus Y37. The CMCase enzyme was purified to homogeneity by 20.4-fold with 21.73% recovery using single-step hydrophobic interaction chromatography and biochemically characterized. CMCase showed a molecular weight of approximately 50 kDa as determined by SDS-PAGE. The activity profile of the CMCase enzyme exhibited optimum activity at 45 degrees C and pH 5.0. The activity was highly stable at alkaline pH levels. More than 90% of the original CMCase activity was maintained at relatively high temperatures ranging from 55 to 65 degrees C. The enzyme activity was induced by Ca2+, Cd2+, CO2+, K+, Mg2+, and Na1+, whereas it was strongly inhibited by phenylmethanesulfonyl fluoride and iodoacetic acid. The enzyme tolerated Hg2+ up to 10 mM and presented hydrolytic activity towards glucan, filter paper, laminarin, and CMC but not o-nitrophenyl beta-D-galactopyranoside. Kinetic analysis of the purified enzyme showed K-m and V-max values of 0.19 mg mL(-1) and 7.46 U mL(-1), respectively. The biochemical properties of this CMCase make the enzyme a good candidate for many industrial applications.