Expression, purification and characterization of meta-cleavage enzyme CarBaBb froim Novosphiongobium sp. K
The meta-cleavage enzyme carbabb of carbazole-degrader novosphiongobium sp. Ka1 werecloned, expressed and purified to homogeneity in escherichia coli strain. The enzyme was cloned with 6x histidine residues attached at the c-terminal of large subunit carbb for purification using affinity chromatogra...
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Hibiscus Publisher
2013
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| Online Access: | http://ir.unimas.my/id/eprint/1571/ http://ir.unimas.my/id/eprint/1571/1/zulkharnain.pdf |
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| author | Zulkharnain, A.B Maeda, R. Omori, T. |
| author_facet | Zulkharnain, A.B Maeda, R. Omori, T. |
| author_sort | Zulkharnain, A.B |
| building | UNIMAS Institutional Repository |
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| description | The meta-cleavage enzyme carbabb of carbazole-degrader novosphiongobium sp. Ka1 werecloned, expressed and purified to homogeneity in escherichia coli strain. The enzyme was cloned with 6x histidine residues attached at the c-terminal of large subunit carbb for purification using affinity chromatography methodprior to gel filtration chromatography. The carbabb, a two-subunit meta-cleavage enzyme, approximately 30 kda for carbb dan 10 kda for carba, was found to be α2β2-heterotetrameric (mr 80,000), showed highest activity at ph 8.5 and temperature 30°c. Carbabb showed highest catalytic activity towards 2,3-dihydroxybiphenyl with kcat/km 4.1 m-1s-1, and overall higher catalytic activities towards biphenyl-type substrates in comparison to catechol-type substrates.Based on the similarities, this meta-cleavage enzyme fromnovosphiongobium sp. Ka1 would also be a good candidate for protein crystallization and structural studies apart from carbabb from strain p. Resinovorans strain ca10. |
| first_indexed | 2025-11-15T05:57:34Z |
| format | Article |
| id | unimas-1571 |
| institution | Universiti Malaysia Sarawak |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T05:57:34Z |
| publishDate | 2013 |
| publisher | Hibiscus Publisher |
| recordtype | eprints |
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| spelling | unimas-15712021-04-30T05:03:08Z http://ir.unimas.my/id/eprint/1571/ Expression, purification and characterization of meta-cleavage enzyme CarBaBb froim Novosphiongobium sp. K Zulkharnain, A.B Maeda, R. Omori, T. Q Science (General) QR Microbiology The meta-cleavage enzyme carbabb of carbazole-degrader novosphiongobium sp. Ka1 werecloned, expressed and purified to homogeneity in escherichia coli strain. The enzyme was cloned with 6x histidine residues attached at the c-terminal of large subunit carbb for purification using affinity chromatography methodprior to gel filtration chromatography. The carbabb, a two-subunit meta-cleavage enzyme, approximately 30 kda for carbb dan 10 kda for carba, was found to be α2β2-heterotetrameric (mr 80,000), showed highest activity at ph 8.5 and temperature 30°c. Carbabb showed highest catalytic activity towards 2,3-dihydroxybiphenyl with kcat/km 4.1 m-1s-1, and overall higher catalytic activities towards biphenyl-type substrates in comparison to catechol-type substrates.Based on the similarities, this meta-cleavage enzyme fromnovosphiongobium sp. Ka1 would also be a good candidate for protein crystallization and structural studies apart from carbabb from strain p. Resinovorans strain ca10. Hibiscus Publisher 2013 Article NonPeerReviewed text en http://ir.unimas.my/id/eprint/1571/1/zulkharnain.pdf Zulkharnain, A.B and Maeda, R. and Omori, T. (2013) Expression, purification and characterization of meta-cleavage enzyme CarBaBb froim Novosphiongobium sp. K. Journal of biochemistry, microbiology and biotechnology, 1 (1). pp. 11-16. ISSN 2289-5779 http://journal.hibiscuspublisher.com/ |
| spellingShingle | Q Science (General) QR Microbiology Zulkharnain, A.B Maeda, R. Omori, T. Expression, purification and characterization of meta-cleavage enzyme CarBaBb froim Novosphiongobium sp. K |
| title | Expression, purification and characterization of meta-cleavage enzyme CarBaBb froim Novosphiongobium sp. K |
| title_full | Expression, purification and characterization of meta-cleavage enzyme CarBaBb froim Novosphiongobium sp. K |
| title_fullStr | Expression, purification and characterization of meta-cleavage enzyme CarBaBb froim Novosphiongobium sp. K |
| title_full_unstemmed | Expression, purification and characterization of meta-cleavage enzyme CarBaBb froim Novosphiongobium sp. K |
| title_short | Expression, purification and characterization of meta-cleavage enzyme CarBaBb froim Novosphiongobium sp. K |
| title_sort | expression, purification and characterization of meta-cleavage enzyme carbabb froim novosphiongobium sp. k |
| topic | Q Science (General) QR Microbiology |
| url | http://ir.unimas.my/id/eprint/1571/ http://ir.unimas.my/id/eprint/1571/ http://ir.unimas.my/id/eprint/1571/1/zulkharnain.pdf |