title: Involvement of phosphoesterases in tributyl phosphate degradation in Sphingobium sp. strain RSMS.
abstract: A tri- and dibutyl phosphate (TBP/DBP) non-degrading spontaneous mutant, Sphingobium SS22, was derived from the Sphingobium sp. strain RSMS (wild type). Unlike the wild type strain, Sphingobium SS22 could not grow in a minimal medium supplemented with TBP or DBP as the sole source of carbon or phosphorous. Sphingobium SS22 also did not form any of the intermediates or end products of TBP or DBP degradation, namely DBP, butanol or inorganic phosphate. Proteomic analysis revealed the absence of three prominent proteins in Sphingobium SS22 as compared to wild type. These proteins were identified by MALDI mass spectrometry, and they showed similarities to phosphohydrolase- and exopolyphosphatase-like proteins from other bacteria, which belong to the class of phosphoesterases. Cellular proteins of Sphingobium SS22 showed none or negligible phosphodiesterase (PDE) and phosphomonoesterase (PME) activities at pH 7 and displayed approximately five- and approximately twofold less DBP and monobutyl phosphate (MBP) degradation activity, respectively, in comparison to the wild type strain. In-gel zymographic analysis revealed two PDE and PME activity bands in the wild type strain, one of which was absent in the Sphingobium SS22 mutant. The corresponding proteins from the wild type strain could degrade DBP and MBP. The results demonstrate the involvement of phosphoesterase enzymes in the TBP degradation pathway elucidated earlier.
authors.affiliation: Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400085, India. shyams@barc.gov.in.
authors.forename: Shyam Sunder
authors.initials: SS
authors.lastname: Rangu
authors.affiliation: Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400085, India. bbasu@barc.gov.in.
authors.forename: Bhakti
authors.initials: B
authors.lastname: Basu
authors.affiliation: Fuel Reprocessing Division, Bhabha Atomic research Centre, Trombay, Mumbai, 400085, India. palazhyb@gmail.com.
authors.forename: Bindu
authors.initials: B
authors.lastname: Muralidharan
authors.affiliation: Fuel Reprocessing Division, Bhabha Atomic research Centre, Trombay, Mumbai, 400085, India. sct@apsara.barc.ernet.in.
authors.forename: S C
authors.initials: SC
authors.lastname: Tripathi
authors.affiliation: Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400085, India. aptesk@barc.gov.in.
authors.forename: Shree Kumar
authors.initials: SK
authors.lastname: Apte
mesh_terms: D001711:Biotransformation; D002244:Carbon / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry; D010755:Organophosphates / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D020543:Proteome / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D042301:Sphingomonadaceae / Q000737:chemistry / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism*
chemical_list: D003470:Culture Media; D010755:Organophosphates; D020543:Proteome; C065087:di-n-butylphosphoric acid; D002244:Carbon; C009524:tributyl phosphate; D010744:Phosphoric Monoester Hydrolases
keywords: Dibutyl phosphate; Phosphoesterases; Tributyl phosphate