Biochemical and structural characterisation of a haloalkane dehalogenase from a marine Rhodobacteraceae. Issue 9 (5th March 2014)

• Record Type: Journal Article
• Authors: Novak, Halina R.; Sayer, Christopher; Isupov, Michail N.; Gotz, Dorothee; Spragg, Andrew Mearns; Littlechild, Jennifer A.
• Is Part Of: FEBS letters. Volume 588:Issue 9(2014)
• Page Start: 1616
• Page End: 1622
• Publication Date: 2014-03-05
• Subjects: HLD -- haloalkane dehalogenase -- l-HAD -- l-Haloacid dehalogenase -- DhlA -- Xanthobacter autotrophicus haloalkane dehalogenase -- DhaA -- Rhodococcus rhodochrous haloalkane dehalogenase -- DmbC -- Mycobacterium tuberculosis haloalkane dehalogenase -- LinB -- Sphingomonas paucimobilis haloalkane dehalogenase -- DbjA -- Bradyrhizobium japonicum USDA110 haloalkane dehalogenase -- DppA -- Plesiocystis pacifica SIR-1 haloalkane dehalogenase -- DspA -- Stronglocentrotus purouratus haloalkane dehalogenase -- HanR -- Rhodobacteraceae family haloalkane dehalogenase -- 1HO -- 1-hexanol -- BAM -- benzamidine -- PMSF -- phenylmethysulfonyl fluoride -- Haloalkane dehalogenase -- Marine Rhodobacteraceae -- Three-dimensional structure -- Catalytic activity; Biochemistry -- Periodicals; Biophysics -- Periodicals; Molecular biology -- Periodicals; Biochimie -- Périodiques; Biochemistry; Biophysics; Molecular biology; Periodicals; 572.05
• DOI: 10.1016/j.febslet.2014.02.056
• Languages: English
• ISSNs: 0014-5793

Abstract

Abstract : A putative haloalkane dehalogenase has been identified in a marine Rhodobacteraceae and subsequently cloned and over‐expressed in Escherichia coli . The enzyme has highest activity towards the substrates 1, 6‐dichlorohexane, 1‐bromooctane, 1, 3‐dibromopropane and 1‐bromohexane. The crystal structures of the enzyme in the native and product bound forms reveal a large hydrophobic active site cavity. A deeper substrate binding pocket defines the enzyme preference towards substrates with longer carbon chains. Arg136 at the bottom of the substrate pocket is positioned to bind the distal halogen group of extended di‐halogenated substrates. Abstract : Isolation of a marine Rhodobacteraceae sp. from a tube worm. Identification of a putative haloalkane dehalogenase enzyme. Characterisation and crystal structure of the native haloalkane dehalogenase enzyme and product bound complex. Relationship between the substrate specificity and the size and charge distribution within the active site cavity.