Alkaline active cyanide dihydratase of Flavobacterium indicum MTCC 6936: Growth optimization, purification, characterization and in silico analysis

作者：Kumar Virender; Kumar Vijay; Bhalla Tek Chand^*

来源：International Journal of Biological Macromolecules, 2018, 116: 591-598.

DOI：10.1016/j.ijbiomac.2018.05.075

• Cyanide dihydratase
• F. indicum
• MALDI-TOF
• Homology modeling

The present work explores a rare cyanide dihydratase of Flavobacterium indicum MTCC 6936 for its potential of cyanide degradation. The enzyme is purified to 12 fold with a yield of 76%. SDS and native-PAGE analysis revealed that enzyme was monomer of 40 kDa size. The enzyme works well in mesophilic range at wide array of pH. The thermostability profile of cyanide dihydratase revealed that the enzyme is quite stable at 30 degrees C and 35 degrees C with half-life of 6 h 30 min and 5 h respectively. K-m and V-max for cyanide dihydratase of F. indicum was measured to be 4.76 mM and 45 U mg(-1) with k(cat) calculated to be 273 s(-1) and specificity constant (k(cat)/K-m) to be around 5.67 mM(-1) s(-1). MALDI-TOF analysis of purified protein revealed that the amino acid sequence has 50% and 43% sequence identity with putative amino acid sequence of F. indicum and earlier reported cyanide dihydratase of Bacillus pumilus respectively. Homology modeling studies of cyanide dihydratase of F. indicum predicted the catalytic triad of the enzyme indicating Cys at 164, Glu at 46 and Lys at 130th position. The purified enzyme has potential applications in bioremediation and analytical sector.

• 出版日期2018-9