Cover Image

Cloning and functional analysis of the genes coding for 4-Aminobenzenesulfonate 3,4-dioxygenase from hydrogenophaga Sp Pbc

The gene coding for the oxygenase component, sadA, of 4-aminobenzenesulfonate (4-ABS) 3,4-dioxygenase in Hydrogenophaga sp. PBC was previously identified via transposon mutagenesis. Expression of wild-type sadA in trans restored the ability of the sadA mutant to grow on 4-ABS. The inclusion of sadB...

Full description

Saved in:
Bibliographic Details
Main Authors: Han, Ming Gan, Shahir, Shafinaz, Yahya, Adibah
Format: Article
Published: Society for General Microbiology 2012
Subjects:
Q Science
Online Access:http://eprints.utm.my/id/eprint/33005/
http://dx.doi.org/10.1099/mic.0.059550-0
Tags: Add Tag
No Tags, Be the first to tag this record!
id my.utm.33005
record_format eprints
spelling my.utm.330052019-03-05T02:03:42Z http://eprints.utm.my/id/eprint/33005/ Cloning and functional analysis of the genes coding for 4-Aminobenzenesulfonate 3,4-dioxygenase from hydrogenophaga Sp Pbc Han, Ming Gan Shahir, Shafinaz Yahya, Adibah Q Science The gene coding for the oxygenase component, sadA, of 4-aminobenzenesulfonate (4-ABS) 3,4-dioxygenase in Hydrogenophaga sp. PBC was previously identified via transposon mutagenesis. Expression of wild-type sadA in trans restored the ability of the sadA mutant to grow on 4-ABS. The inclusion of sadB and sadD, coding for a putative glutamine-synthetase-like protein and a plant-type ferredoxin, respectively, further improved the efficiency of 4-ABS degradation. Transcription analysis using the gfp promoter probe plasmid showed that sadABD was expressed during growth on 4-ABS and 4-sulfocatechol. Heterologous expression of sadABD in Escherichia coli led to the biotransformation of 4-ABS to a metabolite which shared a similar retention time and UV/vis profile with 4-sulfocatechol. The putative reductase gene sadC was isolated via degenerate PCR and expression of sadC and sadABD in E. coli led to maximal 4-ABS biotransformation. In E. coli, the deletion of sadB completely eliminated dioxygenase activity while the deletion of sadC or sadD led to a decrease in dioxygenase activity. Phylogenetic analysis of SadB showed that it is closely related to the glutamine-synthetase-like proteins involved in the aniline degradation pathway. This is the first discovery, to our knowledge, of the functional genetic components for 4-ABS aromatic ring hydroxylation in the bacterial domain. Society for General Microbiology 2012-08 Article PeerReviewed Han, Ming Gan and Shahir, Shafinaz and Yahya, Adibah (2012) Cloning and functional analysis of the genes coding for 4-Aminobenzenesulfonate 3,4-dioxygenase from hydrogenophaga Sp Pbc. Microbiology-Sgm, 158 (Pt8). pp. 1933-1941. ISSN 1350-0872 http://dx.doi.org/10.1099/mic.0.059550-0 DOI:10.1099/mic.0.059550-0
institution Universiti Teknologi Malaysia
building UTM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Teknologi Malaysia
content_source UTM Institutional Repository
url_provider http://eprints.utm.my/
topic Q Science
spellingShingle Q Science
Han, Ming Gan
Shahir, Shafinaz
Yahya, Adibah
Cloning and functional analysis of the genes coding for 4-Aminobenzenesulfonate 3,4-dioxygenase from hydrogenophaga Sp Pbc
description The gene coding for the oxygenase component, sadA, of 4-aminobenzenesulfonate (4-ABS) 3,4-dioxygenase in Hydrogenophaga sp. PBC was previously identified via transposon mutagenesis. Expression of wild-type sadA in trans restored the ability of the sadA mutant to grow on 4-ABS. The inclusion of sadB and sadD, coding for a putative glutamine-synthetase-like protein and a plant-type ferredoxin, respectively, further improved the efficiency of 4-ABS degradation. Transcription analysis using the gfp promoter probe plasmid showed that sadABD was expressed during growth on 4-ABS and 4-sulfocatechol. Heterologous expression of sadABD in Escherichia coli led to the biotransformation of 4-ABS to a metabolite which shared a similar retention time and UV/vis profile with 4-sulfocatechol. The putative reductase gene sadC was isolated via degenerate PCR and expression of sadC and sadABD in E. coli led to maximal 4-ABS biotransformation. In E. coli, the deletion of sadB completely eliminated dioxygenase activity while the deletion of sadC or sadD led to a decrease in dioxygenase activity. Phylogenetic analysis of SadB showed that it is closely related to the glutamine-synthetase-like proteins involved in the aniline degradation pathway. This is the first discovery, to our knowledge, of the functional genetic components for 4-ABS aromatic ring hydroxylation in the bacterial domain.
format Article
author Han, Ming Gan
Shahir, Shafinaz
Yahya, Adibah
author_facet Han, Ming Gan
Shahir, Shafinaz
Yahya, Adibah
author_sort Han, Ming Gan
title Cloning and functional analysis of the genes coding for 4-Aminobenzenesulfonate 3,4-dioxygenase from hydrogenophaga Sp Pbc
title_short Cloning and functional analysis of the genes coding for 4-Aminobenzenesulfonate 3,4-dioxygenase from hydrogenophaga Sp Pbc
title_full Cloning and functional analysis of the genes coding for 4-Aminobenzenesulfonate 3,4-dioxygenase from hydrogenophaga Sp Pbc
title_fullStr Cloning and functional analysis of the genes coding for 4-Aminobenzenesulfonate 3,4-dioxygenase from hydrogenophaga Sp Pbc
title_full_unstemmed Cloning and functional analysis of the genes coding for 4-Aminobenzenesulfonate 3,4-dioxygenase from hydrogenophaga Sp Pbc
title_sort cloning and functional analysis of the genes coding for 4-aminobenzenesulfonate 3,4-dioxygenase from hydrogenophaga sp pbc
publisher Society for General Microbiology
publishDate 2012
url http://eprints.utm.my/id/eprint/33005/
http://dx.doi.org/10.1099/mic.0.059550-0
_version_ 1643649203697614848
score 13.211869

Similar Items