title: Stereochemistry of LinB-catalyzed biotransformation of δ-HBCD to 1R,2R,5S,6R,9R,10S-pentabromocyclododecanol.
abstract: The haloalkane dehalogenase LinB from Sphingobium indicum B90A converts β-hexachlorocyclohexane (β-HCH), the most persistent HCH stereoisomer, to mono- and dihydroxylated metabolites. Recently, we reported that LinB also transforms α-, β- and γ-hexabromocyclododecanes (HBCDs), which are structurally related to HCHs. Here, we show that LinB catalyzes the hydroxylation of δ-HBCD to two pentabromocyclododecanols (PBCDOHs) and two tetrabromocyclododecadiols (TBCDDOHs). The stereochemistry of this enzymatic transformation was deduced from XRD crystal structure data of the substrate δ-HBCD and α(2)-PBCDOH, one of the biotransformation products. Five stereocenters of δ-HBCD are unchanged but the one at C6 is converted to an alcohol with inversion from S- to R-configuration in a nucleophilic, S(N)2-like substitution reaction. Only α(2)-PBCDOH with the 1R,2R,5S,6R,9R,10S-configuration is obtained but not its enantiomer. With only two of the 64 PBCDOHs formed, these transformations indeed are regio- and stereoselective. A conformational analysis revealed that the triple-turn motive, which is predominant in δ-HBCD and in several other HBCD stereoisomers, is also found in the product. This shows that LinB preferentially converted reactive bromine atoms but not those in the conserved triple-turn motive. The widespread contamination with HCHs triggered the bacterial evolution of dehalogenases which acquired the ability to convert these pollutants and their metabolites. We here demonstrate that LinB of S. indicum also transforms HBCDs regio- and stereoselectively following a similar mechanism.
authors.affiliation: Swiss Federal Institute for Materials Science and Technology (Empa), Laboratory of Analytical Chemistry, Überlandstrasse 129, CH-8600 Dübendorf, Switzerland. norbert.heeb@empa.ch
authors.forename: Norbert V
authors.initials: NV
authors.lastname: Heeb
authors.forename: Daniel
authors.initials: D
authors.lastname: Zindel
authors.forename: Heidi
authors.initials: H
authors.lastname: Graf
authors.forename: Valeria
authors.initials: V
authors.lastname: Azara
authors.forename: W Bernd
authors.initials: WB
authors.lastname: Schweizer
authors.forename: Birgit
authors.initials: B
authors.lastname: Geueke
authors.forename: Hans-Peter E
authors.initials: HP
authors.lastname: Kohler
authors.forename: Peter
authors.initials: P
authors.lastname: Lienemann
mesh_terms: D001711:Biotransformation; D002384:Catalysis; D006842:Hydrocarbons, Brominated / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D009713:Nucleotidyltransferases / Q000378:metabolism*; D012989:Soil Pollutants / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D042301:Sphingomonadaceae / Q000378:metabolism*; D013237:Stereoisomerism
chemical_list: D006842:Hydrocarbons, Brominated; D012989:Soil Pollutants; C089796:hexabromocyclododecane; D009713:Nucleotidyltransferases; C057535:lincosaminide O-nucleotidyltransferase