Project description:The project aims to to understand the response of the lin genes in Sphingobium indicum B90A under the stress of HCH isomers and the metabolites formed during degradation of hexachlorocyclohexane (HCH). Entire cell proteome from Sphingobium indicum B90A was extracted in presence of four HCH isomers. Quantitative proteomics confirmed the constitutive expression of the linA, linB and linC genes of the HCH degradation pathway crucial for the initiation of HCH isomers degradation. LinM and LinN were upregulated in the presence of β- and δ-isomers suggested the important role of ABC transporter system in the depletion of β- and δ-HCH. Besides this HCH isomers induced oxidative stress caused systemic changes in strain B90A proteome.
Project description:Genome sequencing of a gamma-hexachlorocyclohexane (g-HCH)-degrading bacterium Sphingobium japonicum strain L15T isolated from an experimental field soil contaminated with g-HCH.
Project description:Among sphingomonads, Sphingobium indicum B90A is widely investigated for its ability to degrade a manmade pesticide, γ-hexachlorocyclohexane (γ-HCH) and its isomers (α-, β-, δ-, and ε-HCH). In this study, complete genome of strain B90A was constructed using Single Molecule Real Time Sequencing (SMRT) and Illumina platform. The complete genome revealed that strain B90A harbors four replicons: one chromosome (3,654,322 bp) and three plasmids designated as pSRL1 (139,218 bp), pSRL2 (108,430 bp) and pSRL3 (43,761 bp). The study determined the precise location of lin genes (genes associated with the degradation of HCH isomers), for example, linA2, linB, linDER, linF, linGHIJ, and linKLMN on the chromosome; linA1, linC, and linF on pSRL1 and linDEbR on pSRL3. Strain B90A contained 26 copies of IS6100 element and most of them (15 copies) was found to be associated with lin genes. Duplication of several lin genes including linA, linDER, linGHIJ, and linF along with two variants of linE, that is, linEa (hydroquinone 1,2-dioxygenase) and linEb (chlorohydroquinone/hydroquinone 1,2-dioxygenase) were identified. This suggests that strain B90A not only possess efficient machinery for upper and lower HCH degradation pathways but it can also act on both hydroquinone and chlorohydroquinone metabolites produced during γ-HCH degradation. Synteny analysis revealed the duplication and transposition of linA gene (HCH dehydrochlorinase) between the chromosome and pSRL1, possibly through homologous recombination between adjacent IS6100 elements. Further, in silico analysis and laboratory experiments revealed that incomplete tyrosine metabolism was responsible for the production of extracellular brown pigment which distinguished strain B90A from other HCH degrading sphingomonads. The precise localization of lin genes, and transposable elements (IS6100) on different replicons now opens up several experimental avenues to elucidate the functions and regulatory mechanism of lin genes acquisition and transfer that were not completely known among the bacterial population inhabiting the HCH contaminated environment.