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

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

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Official URL: http://dx.doi.org/10.1099/mic.0.059550-0

Abstract

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.

Item Type:Article
Uncontrolled Keywords:aminobenzenesulfonate, glutamine-synthetase
Subjects:Q Science
Divisions:Biosciences and Bioengineering (Formerly known)
ID Code:33005
Deposited By: Fazli Masari
Deposited On:31 Jul 2013 16:27
Last Modified:05 Mar 2019 10:03

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