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Volume 149, Issue 11

3- and 4-alkylphenol degradation pathway in sp. strain KL28: genetic organization of the gene cluster and substrate specificities of phenol hydroxylase and catechol 2,3-dioxygenase Free

Abstract

The enzymes and genes responsible for the catabolism of higher alkylphenols have not been characterized in aerobic bacteria. sp. strain KL28 can utilize a wide range of alkylphenols, which include the 4--alkylphenols (C–C). The genes, designated as (for ong-chain lkyphenols), encoding enzymes for the catabolic pathway were cloned from chromosomal DNA and sequenced. The genes are located in a 13·2 kb region with 14 ORFs in the order and with the same transcriptional orientation. The gene is transcribed independently and encodes a member of the XylR/DmpR positive transcriptional regulators. , the first gene in the operon, encodes catechol 2,3-dioxygenase (C23O). The and genes encode a multicomponent phenol hydroxylase (mPH) and enzymes that degrade derivatives of 2-hydroxymuconic semialdehyde (HMS) to TCA cycle intermediates, respectively. The promoter contains motifs at positions −24(GG) and −12(GC) which are typically found in -dependent promoters. A promoter assay using a  : :  transcriptional fusion plasmid showed that promoter activity is inducible and that it responds to a wide range of (alkyl)phenols. The structural genes encoding enzymes required for this catabolism are similar (42–69 %) to those encoded on a catabolic pVI150 plasmid from an archetypal phenol degrader, sp. CF600. However, the locus does not include genes encoding HMS hydrolase and ferredoxin. The latter is known to be functionally associated with C23O for use of 4-alkylcatechols as substrates. The arrangement of the catabolic genes is not commonly found in other -cleavage operons. Substrate specificity studies show that mPH preferentially oxidizes 3- and 4-alkylphenols to 4-alkylcatechols. C23O preferentially oxidizes 4-alkylcatechols via proximal (2,3) cleavage. This indicates that these two key enzymes have unique substrate preferences and lead to the establishment of the initial steps of the lap pathway in strain KL28.

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Keyword(s): C12O, catechol 1,2-dioxygenase , C23O, catechol 2,3-dioxygenase , GFP, green fluorescent protein , HMS, 2-hydroxymuconic semialdehyde , IHF, integration host factor , lap, long-chain alkylphenol and mPH, multicomponent phenol hydroxylase
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2003-11-01
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3- and 4-alkylphenol degradation pathway in Pseudomonas sp. strain KL28: genetic organization of the lap gene cluster and substrate specificities of phenol hydroxylase and catechol 2,3-dioxygenase
Microbiology 149, 3265 (2003); https://doi.org/10.1099/mic.0.26628-0
/content/journal/micro/10.1099/mic.0.26628-0
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