A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä
Expression, purification, characterization and in silico analysis of newly isolated hydrocarbon degrading bleomycin resistance dioxygenase (2020)


Sharma, V., Kumar, R., Sharma, V. K., Yadav, A. K., Tiirola, M., & Sharma, P. K. (2020). Expression, purification, characterization and in silico analysis of newly isolated hydrocarbon degrading bleomycin resistance dioxygenase. Molecular Biology Reports, 47(1), 533-544. https://doi.org/10.1007/s11033-019-05159-x


JYU-tekijät tai -toimittajat


Julkaisun tiedot

Julkaisun kaikki tekijät tai toimittajat: Sharma, Vinay; Kumar, Rajender; Sharma, Vishal Kumar; Yadav, Ashok kumar; Tiirola, Marja; Sharma, Pushpender Kumar

Lehti tai sarja: Molecular Biology Reports

ISSN: 0301-4851

eISSN: 1573-4978

Julkaisuvuosi: 2020

Volyymi: 47

Lehden numero: 1

Artikkelin sivunumerot: 533-544

Kustantaja: Springer Netherlands

Julkaisumaa: Alankomaat

Julkaisun kieli: englanti

DOI: https://doi.org/10.1007/s11033-019-05159-x

Julkaisun avoin saatavuus: Ei avoin

Julkaisukanavan avoin saatavuus:


Tiivistelmä

In the present investigation, we report cloning, expression, purification and characterization of a novel Bleomycin Resistance Dioxygenase (BRPD). His-tagged fusion protein was purified to homogeneity using Ni-NTA affinity chromatography, yielding 1.2 mg of BRPD with specific activity of 6.25 U mg−1 from 600 ml of E. coli culture. Purified enzyme was a dimer with molecular weight ~ 26 kDa in SDS-PAGE and ~ 73 kDa in native PAGE analysis. The protein catalyzed breakdown of hydrocarbon substrates, including catechol and hydroquinone, in the presence of metal ions, as characterized via spectrophotometric analysis of the enzymatic reactions. Bleomycin binding was proven using the EMSA gel retardation assay, and the putative bleomycin binding site was further determined by in silico analysis. Molecular dynamic simulations revealed that BRPD attains octahedral configuration in the presence of Fe2+ ion, forming six co-ordinate complexes to degrade hydroquinone-like molecules. In contrary, in the presence of Zn2+ ion BRPD adopts tetrahedral configuration, which enables degradation of catechol-like molecules.


YSO-asiasanat: biologinen puhdistus; entsyymit; oksidoreduktaasit; haitalliset aineet; torjunta-aineet; aromaattiset hiilivedyt; molekyylidynamiikka

Vapaat asiasanat: metagenomics; dioxygenase; aromatic hydrocarbons; pesticides; pollutant; molecular modelling


Liittyvät organisaatiot


OKM-raportointi: Kyllä

Raportointivuosi: 2020

JUFO-taso: 1


Viimeisin päivitys 2021-29-07 klo 08:08