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Expression, purification and characterization of extradiol dioxygenase carbb involved in carbazole degradation pathway

Extradiol dioxygenase is a family of enzymes essential for ring cleavage reactions in aromatic compounds degradation pathways. Extradiol dioxygenases belonging to the majority of aromatic compound degradation pathways are single peptide proteins, while a small subset of in this family was reported t...

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Main Authors: Maliki, Intan Mariana, Abdul Manas, Nor Hasmaliana, Ahmad, Siti Aqlima, Zulkharnain, Azham
Format: Article
Published: Malaysian Society for Biochemistry and Molecular Biology 2021
Subjects:
Q Science (General)
Online Access:http://eprints.utm.my/id/eprint/94983/
https://msbmb2010.wixsite.com/mjbmb/april-2021
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spelling my.utm.949832022-04-29T22:32:36Z http://eprints.utm.my/id/eprint/94983/ Expression, purification and characterization of extradiol dioxygenase carbb involved in carbazole degradation pathway Maliki, Intan Mariana Abdul Manas, Nor Hasmaliana Ahmad, Siti Aqlima Zulkharnain, Azham Q Science (General) Extradiol dioxygenase is a family of enzymes essential for ring cleavage reactions in aromatic compounds degradation pathways. Extradiol dioxygenases belonging to the majority of aromatic compound degradation pathways are single peptide proteins, while a small subset of in this family was reported to be two subunits complex proteins. The extradiol dioxygenase CarB protein is a protein complex consisting of catalytic subunit CarBb with a smaller subunit CarBa. This enzyme was reported to show no ring cleavage activity without the expression of both peptides. However, to date, there was no specific study to confirm CarBb protein dependency on CarBa protein for its ring cleavage activity. In this study, we cloned, heterologously expressed and purified CarBb in E. coli. CarBb protein showed appreciable ring cleavage activity without expression of CarBa protein. The Km and Vmax values calculated were 163.68 μM and 1.19 μM/min. The effects of pH and temperature suggested that the CarBb protein was significantly unstable, suggesting that the CarBa protein may be responsible for the structural stability of the CarBb protein to function as an effective ring cleavage enzyme. Malaysian Society for Biochemistry and Molecular Biology 2021-04 Article PeerReviewed Maliki, Intan Mariana and Abdul Manas, Nor Hasmaliana and Ahmad, Siti Aqlima and Zulkharnain, Azham (2021) Expression, purification and characterization of extradiol dioxygenase carbb involved in carbazole degradation pathway. Malaysian Journal of Biochemistry and Molecular Biology, 24 (1). pp. 77-82. ISSN 1511-2616 https://msbmb2010.wixsite.com/mjbmb/april-2021
institution Universiti Teknologi Malaysia
building UTM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Teknologi Malaysia
content_source UTM Institutional Repository
url_provider http://eprints.utm.my/
topic Q Science (General)
spellingShingle Q Science (General)
Maliki, Intan Mariana
Abdul Manas, Nor Hasmaliana
Ahmad, Siti Aqlima
Zulkharnain, Azham
Expression, purification and characterization of extradiol dioxygenase carbb involved in carbazole degradation pathway
description Extradiol dioxygenase is a family of enzymes essential for ring cleavage reactions in aromatic compounds degradation pathways. Extradiol dioxygenases belonging to the majority of aromatic compound degradation pathways are single peptide proteins, while a small subset of in this family was reported to be two subunits complex proteins. The extradiol dioxygenase CarB protein is a protein complex consisting of catalytic subunit CarBb with a smaller subunit CarBa. This enzyme was reported to show no ring cleavage activity without the expression of both peptides. However, to date, there was no specific study to confirm CarBb protein dependency on CarBa protein for its ring cleavage activity. In this study, we cloned, heterologously expressed and purified CarBb in E. coli. CarBb protein showed appreciable ring cleavage activity without expression of CarBa protein. The Km and Vmax values calculated were 163.68 μM and 1.19 μM/min. The effects of pH and temperature suggested that the CarBb protein was significantly unstable, suggesting that the CarBa protein may be responsible for the structural stability of the CarBb protein to function as an effective ring cleavage enzyme.
format Article
author Maliki, Intan Mariana
Abdul Manas, Nor Hasmaliana
Ahmad, Siti Aqlima
Zulkharnain, Azham
author_facet Maliki, Intan Mariana
Abdul Manas, Nor Hasmaliana
Ahmad, Siti Aqlima
Zulkharnain, Azham
author_sort Maliki, Intan Mariana
title Expression, purification and characterization of extradiol dioxygenase carbb involved in carbazole degradation pathway
title_short Expression, purification and characterization of extradiol dioxygenase carbb involved in carbazole degradation pathway
title_full Expression, purification and characterization of extradiol dioxygenase carbb involved in carbazole degradation pathway
title_fullStr Expression, purification and characterization of extradiol dioxygenase carbb involved in carbazole degradation pathway
title_full_unstemmed Expression, purification and characterization of extradiol dioxygenase carbb involved in carbazole degradation pathway
title_sort expression, purification and characterization of extradiol dioxygenase carbb involved in carbazole degradation pathway
publisher Malaysian Society for Biochemistry and Molecular Biology
publishDate 2021
url http://eprints.utm.my/id/eprint/94983/
https://msbmb2010.wixsite.com/mjbmb/april-2021
_version_ 1732945417780330496
score 13.211869

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