title: Biochemical characterization of two haloalkane dehalogenases: DccA from Caulobacter crescentus and DsaA from Saccharomonospora azurea.
abstract: Two putative haloalkane dehalogenases (HLDs) of the HLD-I subfamily, DccA from Caulobacter crescentus and DsaA from Saccharomonospora azurea, have been identified based on sequence comparisons with functionally characterized HLD enzymes. The two genes were synthesized, functionally expressed in E. coli and shown to have activity toward a panel of haloalkane substrates. DsaA has a moderate activity level and a preference for long (greater than 3 carbons) brominated substrates, but little activity toward chlorinated alkanes. DccA shows high activity with both long brominated and chlorinated alkanes. The structure of DccA was determined by X-ray crystallography and was refined to 1.5 Å resolution. The enzyme has a large and open binding pocket with two well-defined access tunnels. A structural alignment of HLD-I subfamily members suggests a possible basis for substrate specificity is due to access tunnel size.
authors.affiliation: Department of Chemistry, Hofstra University, Hempstead, New York, 11549.
authors.forename: Lauren
authors.initials: L
authors.lastname: Carlucci
authors.affiliation: Department of Chemistry, Hofstra University, Hempstead, New York, 11549.
authors.forename: Edward
authors.initials: E
authors.lastname: Zhou
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, 10461.
authors.forename: Vladimir N
authors.initials: VN
authors.lastname: Malashkevich
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, 10461.
authors.forename: Steven C
authors.initials: SC
authors.lastname: Almo
authors.affiliation: Department of Chemistry, Hofstra University, Hempstead, New York, 11549.
authors.forename: Emily C
authors.initials: EC
authors.lastname: Mundorff
mesh_terms: D039903:Actinobacteria / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D016935:Caulobacter crescentus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D018360:Crystallography, X-Ray; D006867:Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D011487:Protein Conformation; D040681:Structural Homology, Protein; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D006867:Hydrolases; C046137:haloalkane dehalogenase
keywords: Caulobacter crescentus; DccA; HLD-I subfamily; Saccharomonospora azurea; haloalkane dehalogenase; substrate specificity
references.citation: Nucleic Acids Res. 2002 Apr 15;30(8):E35-5
references.pmid: 11937643
references.citation: Appl Environ Microbiol. 2002 Jul;68(7):3582-7
references.pmid: 12089046
references.citation: Appl Environ Microbiol. 2002 Aug;68(8):3724-30
references.pmid: 12147465
references.citation: J Biol Chem. 2003 Dec 26;278(52):52622-8
references.pmid: 14525993
references.citation: Biotechnol Bioeng. 2004 Apr 5;86(1):47-54
references.pmid: 15007840
references.citation: Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
references.pmid: 15299926
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2256-68
references.pmid: 15572779
references.citation: Biotechnol Bioeng. 2005 Aug 5;91(3):304-13
references.pmid: 15929125
references.citation: Nucleic Acids Res. 2006 Jul 1;34(Web Server issue):W116-8
references.pmid: 16844972
references.citation: Proteins. 2007 May 1;67(2):305-16
references.pmid: 17295320
references.citation: Biochemistry. 2007 Aug 14;46(32):9239-49
references.pmid: 17645312
references.citation: Plant Physiol. 2008 Jul;147(3):1192-8
references.pmid: 18467461
references.citation: J Environ Manage. 2009 Jun;90(8):2841-4
references.pmid: 19237237
references.citation: Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Apr 1;65(Pt 4):353-6
references.pmid: 19342778
references.citation: J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674
references.pmid: 19461840
references.citation: Appl Environ Microbiol. 2009 Aug;75(15):5157-60
references.pmid: 19502442
references.citation: Nat Chem Biol. 2009 Oct;5(10):727-33
references.pmid: 19701186
references.citation: PLoS One. 2009 Sep 14;4(9):e7002
references.pmid: 19759823
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21
references.pmid: 20057044
references.citation: Biochem J. 2011 Apr 15;435(2):345-54
references.pmid: 21294712
references.citation: Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):235-42
references.pmid: 21460441
references.citation: Appl Microbiol Biotechnol. 2011 Aug;91(4):1049-60
references.pmid: 21603934
references.citation: Methods. 2011 Sep;55(1):94-106
references.pmid: 21821126
references.citation: Environ Sci Technol. 1995 Sep 1;29(9):2339-45
references.pmid: 22280276
references.citation: J Mol Biol. 1990 Oct 5;215(3):403-10
references.pmid: 2231712
references.citation: Appl Environ Microbiol. 2012 Jul;78(14):4995-8
references.pmid: 22582053
references.citation: Biotechnol J. 2013 Jan;8(1):32-45
references.pmid: 22965918
references.citation: PLoS Comput Biol. 2012;8(10):e1002708
references.pmid: 23093919
references.citation: Appl Microbiol Biotechnol. 2014 Oct;98(20):8573-82
references.pmid: 24770384
references.citation: Mol Microbiol. 2015 Aug;97(3):439-53
references.pmid: 25899475
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: Appl Environ Microbiol. 1985 Mar;49(3):673-7
references.pmid: 3994371
references.citation: J Bacteriol. 1985 Aug;163(2):635-9
references.pmid: 4019411
references.citation: Methods Enzymol. 1979;63:103-38
references.pmid: 502857
references.citation: J Biol Chem. 1994 Jul 1;269(26):17490-4
references.pmid: 8021255
references.citation: Nature. 1993 Jun 24;363(6431):693-8
references.pmid: 8515812
references.citation: Appl Microbiol Biotechnol. 1998 Dec;50(6):633-57
references.pmid: 9891928