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1、PB 实验设计及结果因素 编码 水平1 +1酵母浸粉 (g/l) A 8 12烘培豆粉 (g/l) B 8 12酵母粉 (g/l) C 5 7.5MgSO4 (g/l) D 0.1 0.3ZnSO4 (g/l) E 0.1 0.15NH4Cl (g/l)CaCl2 (g/l)F 1G 0.051.50.1K2HPO4 (g/l)吐温-80 (ml/l)2 个虚拟项H 1 I 5310根据单因素试验结果以及前人试验成果，选取酵母浸粉、烘培豆粉、酵母粉、MgSO 4、 ZnSO4、NH 4Cl、CaCl 2、K 2HPO4、吐温-80，9 个因素作为筛选因素，从中筛选影响 D-对羟基苯甘氨酸发酵的
2、显著因素。以发酵液中酶活性作为响应值，9 个因素各选高低两个水平，并设置 2 个虚拟值估量误差。表 3-11 Plackett-Burman 试验设计参数和水平代码 因素 低水平（-1） 高水平（+1）A 酵母浸粉 (g/l) 8 12B 烘培豆粉 (g/l) 8 12C 酵母粉 (g/l) 5 7.5D MgSO4 (g/l) 0.1 0.3E ZnSO4 (g/l) 0.1 0.15F NH4Cl (g/l) 1 1.5G CaCl2 (g/l) 0.05 0.1H K2HPO4 (g/l) 1 3I 吐温-80 (ml/l) 5 10J Dummy1 0 0K Dummy2 0 0表 3
3、-12 Plackett-Burman 试验设计与结果序号 A B C D E F G H I J K11 酶活U/mL1 +1 +1 -1 +1 +1 +1 -1 -1 -1 +1 -1 0.2032 -1 +1 +1 -1 +1 +1 +1 -1 -1 -1 +1 0.2513 +1 -1 +1 +1 -1 +1 +1 +1 -1 -1 -1 0.2124 -1 +1 -1 +1 +1 -1 +1 +1 +1 -1 -1 0.1425 -1 -1 +1 -1 +1 +1 -1 +1 +1 +1 -1 0.1376 -1 -1 -1 +1 -1 +1 +1 -1 +1 +1 +1 0.158
4、7 +1 -1 -1 -1 +1 -1 +1 +1 -1 +1 +1 0.1248 +1 +1 -1 -1 -1 +1 -1 +1 +1 -1 +1 0.1649 +1 +1 +1 -1 -1 -1 +1 -1 +1 +1 -1 0.18810 -1 +1 +1 +1 -1 -1 -1 +1 -1 +1 +1 0.14411 +1 -1 +1 +1 +1 -1 -1 -1 +1 -1 +1 0.16812 -1 -1 -1 -1 -1 -1 -1 -1 -1 -1 -1 0.134根据 Plackett-Burman 试验设计的结果可以看出，所筛选的 9 个因素对响应值的影响。如下表：表 3-
5、13 Plackett-Burman 试验方差分析结果因子 均方和 自由度 均方 F 值 p 值Model 0.015 9 1.635 3102.86 0.2861酵母浸粉 7.208 4101 7.208 4101.26 0.3784烘培豆粉 2.107 31 2.107 33.68 0.1951酵母粉 2.552 1 2.552 4.46 0.1691MgSO4 7.008 501 7.008 5100.12 0.7598ZnSO4 5.208 1 5.208 0.091 0.7914NH4Cl 4.219 31 4.219 37.37 0.1131CaCl2 1.302 01 1.302 102.27 0.2705K2HPO4 2.670 31 2.670 34.66 0.1634吐温 -80 1.027 1 1.027 1.79 0.3124校正总和 1.135 302 5.724 410根据软件 Design-Expert 对数据的分析，显示以上 9 个因素对 D-对羟基苯海因的发酵生产并没有显著影响。初步分析是由于实验体系偏小，在实验过程中，培养基的配制，诱导剂的添加以及酶活的测定等步骤中的一些小小操作误差会对实验产生很大的影响，进而影响实验结果。
