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

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Title: Characterisation of the enzymes involved in the diol synthase metabolic pathway in Pseudomonas aeruginosa.
Authors: Shoja-Chaghervand, Shirin1 (AUTHOR), Castells, Marc1 (AUTHOR), Rabanal, Francesc2 (AUTHOR), Cajal, Yolanda3 (AUTHOR), Manresa, Angeles1 (AUTHOR), Estupiñán, Mónica1 (AUTHOR), Busquets, Montserrat1,4 (AUTHOR) mbusquets@ub.edu
Source: Process Biochemistry. Sep2022, Vol. 120, p301-312. 12p.
Subjects: Inductively coupled plasma mass spectrometry, Pseudomonas aeruginosa, Dioxygenases, Fourier transform spectroscopy, Enzymes, Cellular inclusions
Abstract: This study is the first attempt to report the biochemical characterisation of the 10 S -dioxygenase (10 S -DOX) and 7 S , 10 S -diol synthase (7,10-DS). Both enzymes showed similar pH profiles with 10 S -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 10 S -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: 10 S -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 10 S -DOX in the presence of Mn2+ and Fe2+ was reduced to 51.6 % and 61.8. Aggregated proteins producing 10 S -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. [Display omitted] • Biochemical characterisation of recombinant 10 S -dioxygenase (10 S -DOX) and 7,10-diolsynthase (7,10-DS) from P. aeruginosa. • Production of 10 S- hydroxy(per)-oxi-(8 E)-octadecenoic acid using recombinant 10 S -DOX. • Aggregate proteins 10 S -DOX and 7,10-DS were characterised as inclusion bodies. • Oleic acid conversion by diol syntase system is not restricted to P. aeruginosa. [ABSTRACT FROM AUTHOR]
Copyright of Process Biochemistry is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: Characterisation of the enzymes involved in the diol synthase metabolic pathway in Pseudomonas aeruginosa.
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  Data: <searchLink fieldCode="AR" term="%22Shoja-Chaghervand%2C+Shirin%22">Shoja-Chaghervand, Shirin</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Castells%2C+Marc%22">Castells, Marc</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Rabanal%2C+Francesc%22">Rabanal, Francesc</searchLink><relatesTo>2</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Cajal%2C+Yolanda%22">Cajal, Yolanda</searchLink><relatesTo>3</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Manresa%2C+Angeles%22">Manresa, Angeles</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Estupiñán%2C+Mónica%22">Estupiñán, Mónica</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Busquets%2C+Montserrat%22">Busquets, Montserrat</searchLink><relatesTo>1,4</relatesTo> (AUTHOR)<i> mbusquets@ub.edu</i>
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  Data: <searchLink fieldCode="JN" term="%22Process+Biochemistry%22">Process Biochemistry</searchLink>. Sep2022, Vol. 120, p301-312. 12p.
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  Data: <searchLink fieldCode="DE" term="%22Inductively+coupled+plasma+mass+spectrometry%22">Inductively coupled plasma mass spectrometry</searchLink><br /><searchLink fieldCode="DE" term="%22Pseudomonas+aeruginosa%22">Pseudomonas aeruginosa</searchLink><br /><searchLink fieldCode="DE" term="%22Dioxygenases%22">Dioxygenases</searchLink><br /><searchLink fieldCode="DE" term="%22Fourier+transform+spectroscopy%22">Fourier transform spectroscopy</searchLink><br /><searchLink fieldCode="DE" term="%22Enzymes%22">Enzymes</searchLink><br /><searchLink fieldCode="DE" term="%22Cellular+inclusions%22">Cellular inclusions</searchLink>
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  Data: This study is the first attempt to report the biochemical characterisation of the 10 S -dioxygenase (10 S -DOX) and 7 S , 10 S -diol synthase (7,10-DS). Both enzymes showed similar pH profiles with 10 S -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 10 S -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: 10 S -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 10 S -DOX in the presence of Mn2+ and Fe2+ was reduced to 51.6 % and 61.8. Aggregated proteins producing 10 S -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. [Display omitted] • Biochemical characterisation of recombinant 10 S -dioxygenase (10 S -DOX) and 7,10-diolsynthase (7,10-DS) from P. aeruginosa. • Production of 10 S- hydroxy(per)-oxi-(8 E)-octadecenoic acid using recombinant 10 S -DOX. • Aggregate proteins 10 S -DOX and 7,10-DS were characterised as inclusion bodies. • Oleic acid conversion by diol syntase system is not restricted to P. aeruginosa. [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
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  Data: <i>Copyright of Process Biochemistry is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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RecordInfo BibRecord:
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      – Type: doi
        Value: 10.1016/j.procbio.2022.06.017
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      – Code: eng
        Text: English
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      Pagination:
        PageCount: 12
        StartPage: 301
    Subjects:
      – SubjectFull: Inductively coupled plasma mass spectrometry
        Type: general
      – SubjectFull: Pseudomonas aeruginosa
        Type: general
      – SubjectFull: Dioxygenases
        Type: general
      – SubjectFull: Fourier transform spectroscopy
        Type: general
      – SubjectFull: Enzymes
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      – SubjectFull: Cellular inclusions
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    Titles:
      – TitleFull: Characterisation of the enzymes involved in the diol synthase metabolic pathway in Pseudomonas aeruginosa.
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            NameFull: Shoja-Chaghervand, Shirin
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            – D: 01
              M: 09
              Text: Sep2022
              Type: published
              Y: 2022
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