Sreena C P, Sebastian Denoj
Department of Life Sciences, University of Calicut, Malappuram, Kerala 673635, India.
J Genet Eng Biotechnol. 2018 Jun;16(1):9-16. doi: 10.1016/j.jgeb.2017.12.005. Epub 2018 Jan 4.
The production of cellulase by MU S1, a strain isolated from Eravikulam National Park, was optimized using one-factor-at-a-time (OFAT) and statistical methods. Physical parameters like incubation temperature and agitation speed were optimized using OFAT and found to be 40 °C and 150 rpm, respectively, whereas, medium was optimized by statistical tools. Plackett-Burman design (PBD) was employed to screen the significant variables that highly influence cellulase production. The design showed carboxymethyl cellulose (CMC), yeast extract, NaCl, pH, MgSO and NaNO as the most significant components that affect cellulase production. Among these CMC, yeast extract, NaCl and pH showed positive effect whereas MgSO and NaNO were found to be significant at their lower levels. The optimum levels of the components that positively affect enzyme production were determined using response surface methodology (RSM) based on central composite design (CCD). Three factors namely CMC, yeast extract and NaCl were studied at five levels whilst pH of the medium was kept constant at 7. The optimal levels of the components were CMC (13.46 g/l), yeast extract (8.38 g/l) and NaCl (6.31 g/l) at pH 7. The maximum cellulase activity in optimized medium was 566.66 U/ml which was close to the predicted activity of 541.05 U/ml. Optimization of physical parameters and medium components showed an overall 3.2-fold increase in activity compared to unoptimized condition (179.06 U/ml).
从埃拉维库勒姆国家公园分离出的菌株MU S1所产生的纤维素酶,采用单因素法(OFAT)和统计方法进行了优化。使用单因素法对培养温度和搅拌速度等物理参数进行了优化,结果分别为40℃和150转/分钟,而培养基则通过统计工具进行了优化。采用Plackett-Burman设计(PBD)来筛选对纤维素酶产生有高度影响的显著变量。该设计表明,羧甲基纤维素(CMC)、酵母提取物、NaCl、pH值、MgSO₄和NaNO₃是影响纤维素酶产生的最显著成分。其中,CMC、酵母提取物、NaCl和pH值显示出正向影响,而MgSO₄和NaNO₃在较低水平时具有显著性。基于中心复合设计(CCD)的响应面法(RSM)确定了对酶产生有正向影响的成分的最佳水平。研究了CMC、酵母提取物和NaCl这三个因素在五个水平下的情况,同时将培养基的pH值保持在7不变。在pH值为7时,各成分的最佳水平分别为CMC(13.46克/升)、酵母提取物(8.38克/升)和NaCl(6.31克/升)。优化培养基中的最大纤维素酶活性为566.66 U/ml,接近预测活性541.05 U/ml。与未优化条件(179.06 U/ml)相比,物理参数和培养基成分的优化使活性总体提高了3.2倍。