Triphenyltin degradation and proteomic response by an engineered Escherichia coli expressing cytochrome P450 enzyme

• Published in: Ecotoxicology and environmental safety 2017-03, Vol.137, p.29-34
• Main Authors: Yi, Wenying, Yang, Kunliang, Ye, Jinshao, Long, Yan, Ke, Jing, Ou, Huase
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Bacillus thuringiensis; Bacillus thuringiensis - genetics; Bacillus thuringiensis - metabolism; Biodegradation, Environmental; Cloning, Molecular; Cytochrome P-450 Enzyme System - genetics; Cytochrome P-450 Enzyme System - metabolism; Electron Transport; Environmental Pollutants - analysis; Environmental Pollutants - chemistry; Escherichia coli - genetics; Escherichia coli - metabolism; Gene clone; ITRAQ; Molecular Sequence Data; Organotin; Organotin Compounds - analysis; Organotin Compounds - chemistry; Proteome - metabolism; Proteomics
• ISSN: 0147-6513; 1090-2414
• DOI: 10.1016/j.ecoenv.2016.11.012

Although triphenyltin (TPT) degradation pathway has been determined, information about the enzyme and protein networks involved was severely limited. To this end, a cytochrome P450 hydroxylase (CYP450) gene from Bacillus thuringiensis was cloned and expressed in Escherichia coli BL21 (DE3), namely E. coli pET32a-CYP450, whose dosage at 1gL−1 could degrade 54.6% TPT at 1mgL−1 within 6 d through attacking the carbon-tin bonds of TPT by CYP450. Sequence analysis verified that the CYP450 gene had a 1214bp open reading frame, encoding a protein with 404 amino acids. Proteomic analysis determined that 60 proteins were significantly differentially regulated expression in E. coli pET32a-CYP450 after TPT degradation. The up-regulated proteins enriched in a network related to transport, cell division, biosynthesis of amino acids and secondary metabolites, and microbial metabolism in diverse environments. The current findings demonstrated for the first time that P450 received electrons transferring from NADH could effectively cleave carbon-metal bonds. •P450 could cleave carbon-metal bonds.•PntB regulated electron transfer from NADH to P450 during TPT degradation process.•Up-regulated protein network was related to transport, energy synthesis, cell division.•Up-regulated proteins were related to amino acid synthesis.•TPT did not significantly depressed cellular metabolism at a protein network level.