Dave Bhaumik R, Sudhir Ankit P, Subramanian R B
B. R. Doshi School of Biosciences, Sardar Patel Maidan, Satellite Campus, Vadtal Road, Sardar Patel University, Vallabh-Vidyanagar, Gujarat, India.
Biotechnol Rep (Amst). 2014 Nov 27;6:85-90. doi: 10.1016/j.btre.2014.11.004. eCollection 2015 Jun.
An Endo-cellulase was purified to homogeneity using ammonium sulfate precipitation, ion exchange and size exclusion chromatography from newly isolated strain of RBB-1. The recovery and purification fold were 13.3% and 6.6, respectively, after size exclusion chromatography. The purified cellulase has a molecular mass (M) of 35 kDa. Optimum temperature for the enzyme was found to be 70 °C and stability was upto 80 °C for 1 h. Along with higher stability at 80 °C, enzyme showed half lives of 192 h and 144 h at 50 and 70 °C respectively. The purified cellulase was optimally active at pH 4.0 and was stable over a broad pH range of 3.0-7.0. The enzyme purified showed apparent and values of 37 mg/ml and 82.6 U/min/mg protein respectively with higher salt tolerance of 10% for 1 h.
使用硫酸铵沉淀、离子交换和尺寸排阻色谱法从新分离的RBB - 1菌株中纯化出一种内切纤维素酶，使其达到同质。经过尺寸排阻色谱后，回收率和纯化倍数分别为13.3%和6.6。纯化后的纤维素酶分子量（M）为35 kDa。发现该酶的最适温度为70°C，在80°C下稳定性可达1小时。该酶在80°C时具有较高的稳定性，在50°C和70°C时的半衰期分别为192小时和144小时。纯化后的纤维素酶在pH 4.0时活性最佳，在3.0 - 7.0的较宽pH范围内稳定。纯化后的酶表观Km和Vmax值分别为37 mg/ml和82.6 U/min/mg蛋白质，对10%的盐具有1小时的较高耐受性。
