A novel flavin-containing monooxygenase from Methylophaga sp strain SK1 and its indigo synthesis in Escherichia coli

被引:96
作者
Choi, HS
Kim, JK
Cho, EH
Kim, YC
Kim, JI
Kim, SW [1 ]
机构
[1] Chosun Univ, Res Ctr Proteineous Mat, Kwangju 501759, South Korea
[2] Chosun Univ, Dept Biol Educ, Kwangju 501759, South Korea
[3] Kwangju Inst Sci & Technol, Dept Life Sci, Gwangju 500712, South Korea
关键词
flavin-containing monooxygenase; Methylophaga sp strain SK1; microbial indigo synthesis;
D O I
10.1016/S0006-291X(03)01087-8
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We cloned a gene from Methylophaga sp. strain SKI. This gene was responsible for producing, the blue pigment, indigo. The complete open reading frame was 1371 bp long, which encodes a protein of 456 amino acids. The molecular mass of the encoded protein was 105 kDa, consisting of homodimer of 54 kDa with an isoelectric point of 5.14. The deduced amino acid sequence from the gene showed approximately 30% identities with flavin-containing monooxygenases (FMOs) of human (FMO1-FMO5), Arabidopsis, and yeast. It contained three characteristic sequence motifs of FMOs: FAD binding domain, FMO-identifying sequence motif, and NADPH binding domain. In addition, the biochemical properties such as substrate specificities and absorption spectra were similar to the eukaryotic FMO families. Thus, we assigned the enzyme to be a bacterial FMO. The recombinant Escherichia coli expressing the bacterial FMO produced up to 160 mg of indigo per liter in the tryptophan medium after 12 h cultivation. This suggests that the recombinant strain has a potential to be applied in microbial indigo production. (C) 2003 Elsevier Science (USA). All rights reserved.
引用
收藏
页码:930 / 936
页数:7
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