A cytochrome c from a lupanine-transforming Pseudomonas putida strain is expressed in Escherichia coli during aerobic cultivation and efficiently exported and assembled in the periplasm

被引：3

作者：

Kaderbhai, MA ^{[1
]}

Hopper, DJ ^{[1
]}

Akhtar, KM ^{[1
]}

Abbas, SK ^{[1
]}

Kaderbhai, NN ^{[1
]}

机构：

[1] Univ Wales, Inst Biol Sci, Aberystwyth SY23 3DD, Dyfed, Wales

来源：

APPLIED AND ENVIRONMENTAL MICROBIOLOGY | 2003年 / 69卷 / 08期

关键词：

D O I：

10.1128/AEM.69.8.4727-4731.2003

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

We have cloned, sequenced, and heterologously expressed a periplasmic cytochrome c from a lupanine-utilizing Pseudomonas putida strain. Aerobic batch cultivation of Eseherichia coli TB1 harboring the cytochrome c gene placed downstream of the lac promoter in pUC9 vector resulted in significant production of the holo-cytochrome c in the periplasm (similar to4 mg of hemoprotein/liter of culture). The recombinant cytochrome c was purified to homogeneity and was found to be functional in accepting electrons from lupanine hydroxylase while catalyzing hydroxylation of lupanine. Comparison of the N-terminal amino acid sequence of the isolated cytochrome c with that deduced from the DNA sequence indicated that the signal sequence was processed at the bond position predicted by the SigPep program. The molecular size of the cytochrome c determined by electrospray mass spectrometry (9,595) was in precise agreement with that predicted from the nucleotide sequence.

引用

页码：4727 / 4731

页数：5

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