Literature summary for 3.8.1.5 extracted from

Novak, H.R.; Sayer, C.; Isupov, M.N.; Gotz, D.; Spragg, A.M.; Littlechild, J.A.
Biochemical and structural characterisation of a haloalkane dehalogenase from a marine Rhodobacteraceae (2014), FEBS Lett., 588, 1616-1622.

Cloned(Commentary)

Cloned (Comment)	Organism
HanR, DNA and amino acid sequence determination and analysis, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21-CodonPlus (DE3) Rosetta2	Rhodobacteraceae

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged enzyme free or in complex with substrate 1-bromohexane, microbatch crystallization screening, mixing of 10 mg/ml protein in 0.15 M MgCl2, 0.1 M Tris-HCl, with precipitant 15% PEG 4000, pH 8.0, at 18°C, for substrate complex, soaking the HanR crystal for 1 min in 0.1 M Tris-HCl, 20% PEG 3350, 30% PEG 400, 25 mM 1-bromohexane, pH 4.0, X-ray diffraction structure determination and analysis at 1.61-1.67 A resolution, molecular replacement using the LinB model, PDB ID 1CV2	Rhodobacteraceae

Crystallization (Comment)

Organism

purified recombinant His-tagged enzyme free or in complex with substrate 1-bromohexane, microbatch crystallization screening, mixing of 10 mg/ml protein in 0.15 M MgCl2, 0.1 M Tris-HCl, with precipitant 15% PEG 4000, pH 8.0, at 18°C, for substrate complex, soaking the HanR crystal for 1 min in 0.1 M Tris-HCl, 20% PEG 3350, 30% PEG 400, 25 mM 1-bromohexane, pH 4.0, X-ray diffraction structure determination and analysis at 1.61-1.67 A resolution, molecular replacement using the LinB model, PDB ID 1CV2

Rhodobacteraceae

Organism

Organism	UniProt	Comment	Textmining
Rhodobacteraceae	-	Rhb, isolated from a Polychaeta worm	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminally His-tagged enzyme from Escherichia coli strain BL21-CodonPlus (DE3) Rosetta2 by nickel affinity chromatography, ultrafiltration, and gel filtration	Rhodobacteraceae

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
1,2-dibromoethane + H2O	-	Rhodobacteraceae	2-bromoethanol + bromide	-	?
1,3-dibromopropane + H2O	-	Rhodobacteraceae	3-bromopropanol + bromide	-	?
1,4-dichlorobutane + H2O	-	Rhodobacteraceae	4-chlorobutanol + chloride	-	?
1,6-dichlorohexane + H2O	-	Rhodobacteraceae	6-chlorohexanol + chloride	-	?
1-bromohexane + H2O	-	Rhodobacteraceae	hexanol + bromide	-	?
1-bromooctane + H2O	-	Rhodobacteraceae	octanol + bromide	-	?
2-chloropropane + H2O	-	Rhodobacteraceae	ethanol + chloride	-	?
cyclohexylbromide + H2O	-	Rhodobacteraceae	cyclohexanol + bromide	-	?
cyclohexylchloride + H2O	-	Rhodobacteraceae	cyclohexanol + chloride	-	?
additional information	no activity with 1,2-dichloroethane, 1,2-dichloropropane, and 1,2-dichloro-3-propanol	Rhodobacteraceae	?	-	?

Synonyms

Synonyms	Comment	Organism
HanR	-	Rhodobacteraceae
HLD	-	Rhodobacteraceae

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.2	-	assay at	Rhodobacteraceae

General Information

General Information	Comment	Organism
additional information	the active sites of Han enzymes are characterised by a catalytic pentad. This consists of a catalytic triad and a stabilizing pair of residues. The catalytic triad consists of Asp106 (located between beta-strand 5 and alpha-helix 2), His271 (located between beta-strand 8 and 310 helix 9) and Glu130 (located within beta-strand 6). The side chains of Trp107 and Asn36 bind the substrate halogen, while their main chain nitrogen atoms form the oxyanion hole. The HLD active site is built up of an entrance tunnel and a hydrophobic pocket for substrate binding. The residues lining the substrate pocket mainly belong to the cap domain and define substrate specificity. Minor differences in the substrate pocket can dramatically affect the substrate specificity of very similar HLDs	Rhodobacteraceae