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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my.unimas.ir.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/ |
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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 |
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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. |
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Article |
author |
Zulkharnain, A.B Maeda, R. Omori, T. |
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Zulkharnain, A.B Maeda, R. Omori, T. |
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Zulkharnain, A.B |
title |
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_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_sort |
expression, purification and characterization of meta-cleavage enzyme carbabb froim novosphiongobium sp. k |
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Hibiscus Publisher |
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2013 |
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http://ir.unimas.my/id/eprint/1571/1/zulkharnain.pdf http://ir.unimas.my/id/eprint/1571/ http://journal.hibiscuspublisher.com/ |
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