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2022-06-10

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cc-by (c) Shoja Chaghervand et al., 2022
Si us plau utilitzeu sempre aquest identificador per citar o enllaçar aquest document: https://hdl.handle.net/2445/195944

Characterisation of the enzymes involved in the diol synthase metabolic pathway in Pseudomonas aeruginosa

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Recurs relacionat

https://doi.org/10.1016/j.procbio.2022.06.017

Resum

This study is the first attempt to report the biochemical characterisation of the 10S-dioxygenase (10S-DOX) and 7S, 10S-diol synthase (7,10-DS). Both enzymes showed similar pH profiles with 10S-DOX presenting the highest activity at 30 °C whereas 7,10-DS did not show a clear preferred temperature. These differences were reflected in the thermostability assay, the Km values were 0.89 ± 0.22 mM and 3.26 ± 0.31 mM for 10S-DOX and 7,10-DS, respectively. Inductively coupled plasma mass spectrometry indicated that both enzymes contained bound to haem group Fe2+ as a prosthetic group: 10S-DOX (0.95 mol Fe2+/mol of protein) and 7,10-DS (1.18 mol Fe2+/mol of protein), respectively. Assays using metal cations as cofactors revealed that Mg2+ and Ni2+ enhance 7,10-DS activity, whereas Hg2+ decrease it up to 50 %. The activity of 10S-DOX in the presence of Mn2+ and Fe2+ was reduced to 51.6 % and 61.8. Aggregated proteins producing 10S-DOX and 7,10-DS were characterised as inclusion bodies: IBs-77 and IBs-78 respectively, was performed by Fourier Transform spectroscopy (FT-IR), Atomic Force Microscopy, dye binding, and proteolysis. The specific activity was 1.55 IU/mg for IBs-77 and 1.05 IU/mg for IBs-78. The presence of the oleate-diol synthase pathway in proteobacteria other than Pseudomonas aeruginosa was detected.

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Microscòpia de força atòmica, Temperatura

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Atomic force microscopy, Temperature

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Articles publicats en revistes (Química Inorgànica i Orgànica)
Articles publicats en revistes (Biologia, Sanitat i Medi Ambient)
Articles publicats en revistes (Bioquímica i Biomedicina Molecular)
Articles publicats en revistes (Farmàcia, Tecnologia Farmacèutica i Fisicoquímica)
Articles publicats en revistes (Institut de Nanociència i Nanotecnologia (IN2UB))

Citació

SHOJA CHAGHERVAND, S, CASTELLS, Marc, RABANAL ANGLADA, Francesc, CAJAL VISA, Yolanda, MANRESA PRESAS, Ma. ángeles (maría ángeles), ESTUPIÑÁN, Mónica, BUSQUETS ABIÓ, Montserrat. Characterisation of the enzymes involved in the diol synthase metabolic pathway in Pseudomonas aeruginosa. _Process Biochemistry_. 2022. Vol. 120, núm. 301-312. [consulta: 30 de gener de 2026]. ISSN: 1359-5113. [Disponible a: https://hdl.handle.net/2445/195944]

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