Omer C A, Lenstra R, Litle P J, Dean C, Tepperman J M, Leto K J, Romesser J A, O'Keefe D P
Central Research and Development Department, E.I. Du Pont Co., Wilmington, Delaware 19880.
J Bacteriol. 1990 Jun;172(6):3335-45. doi: 10.1128/jb.172.6.3335-3345.1990.
Streptomyces griseolus ATCC 11796 contains two inducible, herbicide-metabolizing cytochromes P-450 previously designated P-450SU1 and P-450SU2 (P-450CVA1 and P-450CVB1, respectively, using nomenclature of Nebert et al. [D. W. Nebert, M. Adesnik, M. J. Coon, R. W. Estabrook, F. J. Gonzalez, F. P. Guengerich, I. C. Gunsalus, E. F. Johnson, B. Kemper, W. Levin, I. R. Phillips, R. Sato, and M. R. Waterman, DNA 6:1-11, 1987]). Using antibodies directed against cytochrome P-450SU1, its N-terminal amino acid sequence, and amino acid composition, we cloned the suaC gene encoding cytochrome P-450SU1. Similar information about the cytochrome P-450SU2 protein confirmed that a gene cloned by cross-hybridization to the suaC gene was the subC gene encoding cytochrome P-450SU2. The suaC and subC genes were expressed in Escherichia coli, DNA for both genes was sequenced, and the deduced amino acid sequences were compared with that of the well-characterized cytochrome P-450CAM from Pseudomonas putida. Both cytochromes P-450SU1 and P-450SU2 contain several regions of strong similarity with the amino acid sequence of P-450CAM, primarily in regions of the protein responsible for attachment and coordination of the heme prosthetic group.
灰色链霉菌ATCC 11796含有两种可诱导的、代谢除草剂的细胞色素P-450,以前分别命名为P-450SU1和P-450SU2(分别使用Nebert等人的命名法命名为P-450CVA1和P-450CVB1 [D. W. Nebert, M. Adesnik, M. J. Coon, R. W. Estabrook, F. J. Gonzalez, F. P. Guengerich, I. C. Gunsalus, E. F. Johnson, B. Kemper, W. Levin, I. R. Phillips, R. Sato, and M. R. Waterman, DNA 6:1 - 11, 1987])。利用针对细胞色素P-450SU1的抗体、其N端氨基酸序列和氨基酸组成,我们克隆了编码细胞色素P-450SU1的suaC基因。关于细胞色素P-450SU2蛋白的类似信息证实,通过与suaC基因交叉杂交克隆的一个基因是编码细胞色素P-450SU2的subC基因。suaC和subC基因在大肠杆菌中表达,对这两个基因的DNA进行了测序,并将推导的氨基酸序列与恶臭假单胞菌中特征明确的细胞色素P-450CAM的氨基酸序列进行了比较。细胞色素P-450SU1和P-450SU2都含有几个与P-450CAM氨基酸序列高度相似的区域,主要在负责血红素辅基附着和配位的蛋白质区域。