Identification of the flavin monooxygenase responsible for ipso substitution of alkyl and alkoxyphenols in Sphingomonas sp. TTNP3 and Sphingobium xenophagum Bayram

被引:31
作者
Porter, A. W. [1 ]
Campbell, B. R. [1 ]
Kolvenbach, B. A. [2 ]
Corvini, P. F. -X. [2 ]
Benndorf, D. [3 ]
Rivera-Cancel, G. [1 ]
Hay, A. G. [1 ,4 ]
机构
[1] Cornell Univ, Dept Microbiol, New York, NY 10021 USA
[2] Univ Appl Sci NW Switzerland, Sch Life Sci, Inst Ecopreneurship, CH-4132 Muttenz, Switzerland
[3] Otto Von Guericke Univ, D-39106 Magdeburg, Germany
[4] Cornell Univ, Inst Comparat & Environm Toxicol, New York, NY 10021 USA
基金
美国农业部; 美国国家环境保护局;
关键词
Octylphenol monooxygenase; Ipso substitution; Alkylphenol biodegradation; Octylphenol; SP STRAIN TTNP3; NONYLPHENOL ISOMERS; DEGRADATION PATHWAY; METABOLIC PATHWAY; GENE-CLUSTER; OXYGEN-ATOM; BISPHENOL-A; MECHANISM; CYTOCHROME-P450; BIODEGRADATION;
D O I
10.1007/s00253-011-3621-8
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 previously showed that opdA from Sphingomonas sp. PWE1 encodes a putative flavin monooxygenase capable of transforming octylphenol (OP) via type II ipso substitution. Here, we demonstrate that an opdA homolog is responsible for OP and related alkyl/alkoxyphenol degradation in the nonylphenol degrader Sphingomonas sp. TTNP3. PCR and Southern blot analyses revealed that TTNP3 contained an opdA homolog, while a TTNP3 derivative unable to grow on nonylphenol (TTNP3d) did not. OpdA expression was confirmed in wild-type TTNP3 via two dimensional gel electrophoresis. Activity was restored to TTNP3d following complementation with opdA. Sequence analysis of an opdA homolog from another nonylphenol degrader, Sphingobium xenophagum Bayram, revealed that the predicted protein sequences from PWE1 and Bayram were identical, but differed from TTNP3 by four amino acids. In order to assess differences, we heterologously expressed the two unique opdA homologs and compared their effect on the disappearance of five alkyl/alkoxyphenol substrates and subsequent appearance of hydroquinone. For all substrates, except OP, the levels of substrate disappearance and hydroquinone appearance were significantly lower in cultures expressing odpA (TTNP3) than those expressing opdA (PWE1/Bayram). These differences in substrate specificity were consistent with an in silico model which predicted that two of the amino acid differences between odpA (TTNP3) and opdA (PWE1/Bayram) lay in a putative substrate binding pocket. While these strains are known to use the same type II ipso substitution mechanism for alkylphenol degradation, this work provides the first preliminary evidence that opdA homologs also encode the type I ipso substitution activity responsible for the degradation of alkoxyphenols.
引用
收藏
页码:261 / 272
页数:12
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