核苷酸序列的假单胞菌putida细胞色素p - 450凸轮在大肠杆菌基因及其表达。
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昂格尔BP Gunsalus IC, Sligar SG
核苷酸序列的假单胞菌putida细胞色素p - 450凸轮在大肠杆菌基因及其表达。
J生物化学杂志。1986年1月25日,261 (3):1158 - 63。
- PubMed ID
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3003058 (在PubMed]
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细胞色素p - 450凸轮的立体定向亚甲基羟基化催化樟脑形成5-exohydroxycamphor和凸轮上由camC编码基因质粒的假单胞菌putida,写明ATCC 17453。细胞色素p - 450凸轮结构基因突变克隆了互补p . putida(四郎,H。,Rauchfuss B。,Gunsalus i c .学生物化学(1985)。Biophys。Commun >, 130年,412 - 417)。我们报告的完整camC基因的核苷酸序列和155个碱基对的5和3的175个碱基对的旁侧序列。经氨基酸序列比较来自于基因序列的获得纯化蛋白(Haniu, M。回来,l·G。Yasunobu, k . T。、Shastry b。Gunsalus, i c(1982)生物。257年化学,12664 - 12671),5个差异被发现。 The most significant was the addition of a Trp and a Thr residue between Val-54 and Arg-55, thereby increasing the amino acid numbering scheme by 2 after Val-54, bringing the total number of amino acids to 414. Other differences were: Gln-274----Glu-276, Ser-359----His-361, and Asn-405----Asp-407. N-terminal amino acid sequence analysis of the cloned cytochrome P-450cam enzyme expressed in Escherichia coli under the lac promoter showed a faithful translation of the hemo-protein, with the N-terminal Met removed by processing as found in P. putida. Purification to homogeneity of the cloned protein was accomplished by the method used for the CAM plasmid-encoded enzyme of P. putida. The G + C content of the camC gene was found to be 59.0%, caused by a preferred usage of G and C terminated codons. The gene encoding putidaredoxin reductase, camA, was located 22 nucleotides downstream from the cytochrome P-450cam gene. The camA gene initiated with a novel GUG codon, the first such initiator documented in Pseudomonas.