Document Detail

Examination and expansion of the substrate range of m-hydroxybenzoate hydroxylase.
MedLine Citation:
PMID:  18417078     Owner:  NLM     Status:  MEDLINE    
The gene encoding m-hydroxybenzoate hydroxylase (mobA) was cloned from Comamonas testosteroni GZ39. MobA converts m-hydroxybenzoate and to a lesser extent p-hydroxybenzoate to protocatechuate. To explore the structural and functional relationships in phenolic acid monooxygenases, MobA was subjected to in vitro mutagenesis by error-prone PCR and the mutant MobAs were screened for their ability to oxidize phenol or 3-aminophenol. A mutant MobA with a single V257A substitution was able to transform phenol to catechol, providing the first example of monooxygenase acting on phenolic acids that can also hydroxylate phenol. The mutant MobA also has enhanced ability to transform resorcinol, hydroquinone, p-hydroxybenzoate, 2,5-dihydroxybenzoate, 3,4-dihydroxybenzoate, 3-chlorophenol, 4-chlorophenol, 4-chlororesorcinol, and 4-nitrophenol. Several MobA mutants were obtained for their ability to transform 3-aminophenol to a related substituted catechol. Mutant MobAs with single amino acid substitutions (H135P, A400G, or D416A) were derived from these mutants and verified for their ability to transform 3-aminophenol.
Hung-Kuang Chang; Gerben J Zylstra
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Publication Detail:
Type:  Journal Article     Date:  2008-04-14
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  371     ISSN:  1090-2104     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2008 Jun 
Date Detail:
Created Date:  2008-05-09     Completed Date:  2008-06-03     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  149-53     Citation Subset:  IM    
Biotechnology Center for Agriculture and the Environment, School of Environmental and Biological Sciences, Rutgers University, 59 Dudley Road, New Brunswick, NJ 08901-8520, USA.
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MeSH Terms
Amino Acid Sequence
Amino Acid Substitution
Aminophenols / chemistry*
Bacterial Proteins / chemistry*,  genetics,  isolation & purification
Cloning, Molecular
Comamonas testosteroni / enzymology*,  genetics
Directed Molecular Evolution
Mixed Function Oxygenases / chemistry*,  genetics,  isolation & purification
Molecular Sequence Data
Phenol / chemistry*
Protein Conformation
Substrate Specificity
Reg. No./Substance:
0/Aminophenols; 0/Bacterial Proteins; 108-95-2/Phenol; 591-27-5/3-aminophenol; EC 1.-/Mixed Function Oxygenases; EC

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