响应面法优化多粘类芽孢杆菌N10产果胶裂解酶的条件

M. Songpim, P. Vaithanomsat, S. Chuntranuluck
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引用次数: 12

摘要

采用响应面法研究了搅拌速率(X1, 100-300 rpm)、温度(X2, 25-45℃)和pH (X3, 5.5-9.5)对P. polymyxa N10产果胶裂解酶的影响。对酶产率影响最大的因素是温度和ph。拟合得到的二阶多项式回归模型,r2为0.9600 (p < 0.001)。在搅拌速度200转/分、温度35℃、pH 8条件下培养72 h,果胶裂解酶活性达到84.5 U/ml。在5-l搅拌槽生物反应器中,对果胶裂解酶的搅拌速率和曝气进行了优化。在搅拌速度为200 rpm(温度35℃,初始pH为8)时,曝气速率在0.5 ~ 2 vvm范围内变化。在搅拌速度为200 rpm时,剪切力较大,对果胶裂解酶的结构产生负面影响,导致酶活性降低。在生物反应器中,果胶裂解酶的最大活性为110.42 U/ml,与摇瓶发酵的最大值接近。
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Optimization of Pectate Lyase Production from Paenibacillus polymyxa N10 using Response Surface Methodology
The parameters affecting the production of pectate lyase from P. polymyxa N10 were studied using the response surface methodology agitation rate (X1, 100-300 rpm), temperature (X2, 25-45 o C) and pH (X3, 5.5-9.5). The most significant factors influencing enzyme production were temperature and pH. The second order polynomial regression model obtained was fitted and found adequate, with an R 2 of 0.9600 (p < 0.001). A maximum pectate lyase activity of 84.5 U/ml was attained in 72 h of cultivation at agitation rate 200 rpm, temperature 35 o C and pH 8. Optimizations of agitation rate and aeration on pectate lyase production were also carried out in a 5-l stirred-tank bioreactor. The aeration rate was varied in the range of 0.5-2 vvm at agitation rate of 200 rpm (temperature 35 o C and initial pH 8). At agitation rate of 200 rpm, the shear force was high and then decreased the pectate lyase activity due to its negative effect on the enzyme structure. A maximum pectate lyase activity of 110.42 U/ml in the bioreactor was close to that obtained from the shake flask fermentation study.
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