β-甘露聚糖酶发酵液絮凝条件的统计学筛选和响应面优化.docVIP

β-甘露聚糖酶发酵液絮凝条件的统计学筛选响应面优化 韩健，刘朝辉，齐崴，何志敏 （天津大学 化工学院 化学工程研究所，天津 300072） 摘 要: 采用Plackett-Burman（PB）法和中心复合设计（Central Composite Design）法对影响地衣芽孢杆菌TJ-101发酵生产β-甘露聚糖酶粗发酵液絮凝操作的8个条件进行筛选优化。PB实验设计与统计学分析表明：加水量，阳离子聚丙烯酰胺(C-PAM)和阴离子聚丙烯酰胺(A-PAM)的浓度是影响酶活收率的三个关键因素。以酶活收率为响应目标，对三因素进行中心复合设计，并经响应面法优化分析得到影响酶活收率的二阶模型，确定了β-甘露聚糖酶发酵液絮凝实验的最优操作条件为：加水量240.55%(V)，C-PAM 14.13 %(V)，A-PAM 16.97%(V)，絮凝后发酵液的酶活收率达70.42%。 关键词: β-甘露聚糖酶絮凝Plackett-Burman设计，中心复合设计，响应面，优化 : 中图分类号: 文献标识码: A Optimization of the flocculation conditions of β-mannanase broth using statistical screening and response surface methodology HAN Jian, LIU Zhao-hui, QI Wei, HE Zhi-min (Chemical Engineering Research Centre, School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China) Abstract: Plackett-Burman (PB) design and Central Composite Design (CCD) were applied to screening and optimizing 8 factors for the flocculation of β-mannanase production by Bacillus licheniformis TJ-101. The amount of water, and the concentrations of C-PAM and A-PAM, as three key factors, were found to significantly influence the yield of β-mannanase activity via PB design and the following statistic analysis. By CCD design and response surface analysis, the quadratic model for three significant factors was established with β-mannanase activity as the target response. At the optimal flocculation conditions, i.e. 240.55% water, 14.13% C-PAM and 16.97% A-PAM, the yield of enzyme activity achieved 70.42%. Keywords: β-Mannanase; flocculation; Plackett-Burman; central composite design; response surface methodology; optimization β-甘露聚糖酶（1,4-β-D-mannanase, EC 3.2.1.78）是一类能够随机裂解β-1,4糖苷键的半纤维素内切水解酶[1]，可通过多种产酶微生物如芽孢杆菌[2-3]、曲霉[4]等经发酵培养制得。随着人们对自然界半纤维素资源的开发和甘露寡糖药用价值的发现，β-甘露聚糖酶已成为一种极其重要的工业酶制剂，在食品、医药、造纸、纺织、印染、石油开采及生物技术等诸多领域得到广泛应用[5]。 絮凝是生物技术下游加工过程特别是对微生物发酵液进行分离纯化以制备具有实用价值和竞争力产品的重要手段[6-9]，它涉及各种絮凝剂的组合混凝作用和絮凝操作条件的优化，以达到去除发酵液中菌体等杂质，提高产品纯度的目的。 在实验条件优化方面，传统的单因素实验方法费时费力，而且没有考虑因素的交互作用致使最优值并不可靠[10]。Plackett-Burman（PB）[