New Metabolites in the Degradation of α- and γ-Hexachlorocyclohexane (HCH): Pentachlorocyclohexenes Are Hydroxylated to Cyclohexenols and Cyclohexenediols by the Haloalkane Dehalogenase LinB from Sphingobium indicum B90A

• Published in: Journal of agricultural and food chemistry Vol. 56; no. 15; pp. 6594 - 6603
• Main Authors: Raina, Vishakha, Rentsch, Daniel, Geiger, Thomas, Sharma, Poonam, Buser, Hans Rudolf, Holliger, Christof, Lal, Rup, Kohler, Hans-Peter E.
• Format: Journal Article
• Language: English
• Published: WASHINGTON American Chemical Society 13.08.2008; Amer Chemical Soc
• Subjects: Agriculture; Agriculture, Multidisciplinary; biochemical pathways; biodegradation; bioremediation; Carrier Proteins; Chemistry; Chemistry, Applied; cyclohexendiols; cyclohexenes; cyclohexenols; dehalogenation; dehydrochlorinase; dehydrohalogenation; Environmental Chemistry; enzyme substrates; Escherichia coli; Escherichia coli Proteins; Food Science & Technology; Gas Chromatography-Mass Spectrometry; Gene Expression; gene transfer; halidohydrolase; haloalkane dehalogenase; HCH; HCH (pesticide); hydrolases; Hydrolases - genetics; Hydrolases - metabolism; Hydroxylation; Life Sciences & Biomedicine; LinA; LinB; lindane; Lindane - metabolism; Magnetic Resonance Spectroscopy; metabolites; PCCH; pentachlorocyclohexene; pentachlorocyclohexenes; Physical Sciences; Recombinant Proteins - metabolism; Science & Technology; soil pollution; Sphingobium indicum; Sphingobium indicumB90A; Sphingomonadaceae - enzymology; Sphingomonas; stereoisomers; Substrate Specificity; tetrachlorcyclohexenol; trichlorocyclohexenediol; trichlorocyclohexenediol; LinA; LinB; haloalkane dehalogenase; dehydrohalogenation; hydroxylation; tetrachlorcyclohexenol; halidohydrolase; Sphingobium indicum B90A; pentachlorocyclohexene; HCH; PCCH; dehydrochlorinase; DELTA-HEXACHLOROCYCLOHEXANE; TRANSFORMATION; Sphingobium indicum B90A; BETA-HEXACHLOROCYCLOHEXANE; halidohydrolase; IDENTIFICATION
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf800465q

Technical hexachlorocyclohexane (HCH) and lindane are obsolete pesticides whose former production and use led to widespread contaminations posing serious and lasting health and environmental risks. Out of nine possible stereoisomers, α-, β-, γ-, and δ-HCH are usually present at contaminated sites, and research for a better understanding of their biodegradation has become essential for the development of appropriate remediation technologies. Because haloalkane dehalogenase LinB was recently found responsible for the hydroxylation of β-HCH, δ-HCH, and δ-pentachlorocyclohexene (δ-PCCH), we decided to examine whether β- and γ-PCCH, which can be formed by LinA from α- and γ-HCH, respectively, were also converted by LinB. Incubation of such substrates with Escherichia coli BL21 expressing functional LinB originating from Sphingobium indicum B90A showed that both β-PCCH and γ-PCCH were direct substrates of LinB. Furthermore, we identified the main metabolites as 3,4,5,6-tetrachloro-2-cyclohexene-1-ols and 2,5,6-trichloro-2-cyclohexene-1,4-diols by nuclear magnetic resonance spectroscopy and gas chromatography−mass spectrometry. In contrast to α-HCH, γ-HCH was not a substrate for LinB. On the basis of our data, we propose a modified γ-HCH degradation pathway in which γ-PCCH is converted to 2,5-cyclohexadiene-1,4-diol via 3,4,5,6-tetrachloro-2-cyclohexene-1-ol and 2,5,6-trichloro-2-cyclohexene-1,4-diol.