JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2007, Vol. 31 ›› Issue (06): 48-52.doi: 10.3969/j.jssn.1000-2006.2007.06.011

Previous Articles     Next Articles

Purification and Characterization of Cellulase from Trichoderma reesei

JIANG Hua, YU Zhao-hai   

  1. College of Chemical Engineering Nanjing Forestry University, Nanjing 210037, China
  • Online:2007-12-18 Published:2007-12-18

Abstract: The purification steps, e.g. a two-step ultrafiltration, cationic (CM-Sepharose FF) and anionic (DEAE-Sepharose FF) chromatography and size exclusion (Sephadex G-100) chromatography and the six homogeneous cellulase etc, were used to purify three endoglucanases (EG Ⅰ, EG Ⅱ and EG Ⅲ), two cellobiohydrolases (CBH Ⅰ and CBH Ⅱ) and a (β-glucosidase (GB) according to SDS—PAGE analysis from the culture broth of Trichoderma reesei. The specific activities of EG Ⅰ, EG Ⅱ and EG Ⅲ with carboxymethyl celluloses were estimated to be 176.35, 153.96, 64.22IU/mg, respectively. For CBH I and CBH Ⅱ, their specific activities with microcelluloses were 16.86, 4.82IU/mg, respectively. The GB with salicin was 31.00IU/mg. Their Km were 6.70, 8.46, 13.22, 1.37, 3.46, 2.20mg/mL, respectively. The greater Km of the purified enzyme component of the same class is, the smaller their turnover is. Their molecular weights of EG Ⅰ, EG Ⅱ, EG Ⅲ, CBH I, CBH Ⅱ and GB were 50, 46, 25, 65, 58, 75kDa, respectively.

CLC Number: