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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Malaysian Society for Biochemistry and Molecular Biology
2021
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| Online Access: | http://psasir.upm.edu.my/id/eprint/97237/1/ABSTRACT.pdf |
| _version_ | 1796983368040054784 |
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| 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 |
| collection | UPM |
| 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. |
| first_indexed | 2024-03-06T11:05:32Z |
| format | Article |
| id | upm.eprints-97237 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T11:05:32Z |
| publishDate | 2021 |
| publisher | Malaysian Society for Biochemistry and Molecular Biology |
| record_format | dspace |
| spelling | upm.eprints-972372022-09-12T08:43:41Z http://psasir.upm.edu.my/id/eprint/97237/ 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 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 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/97237/1/ABSTRACT.pdf 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). 77 - 82. ISSN 2600-9005 https://shibaura.pure.elsevier.com/en/publications/expression-purification-and-characterization-of-extradiol-dioxyge |
| spellingShingle | 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 |
| title | 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_short | 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 |
| url | http://psasir.upm.edu.my/id/eprint/97237/1/ABSTRACT.pdf |
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