Investigation of the prevalence and catalytic activity of rubredoxin-fused alkane monooxygenases (AlkBs)
Interest in understanding the environmental distribution of the alkane monooxygenase (AlkB) enzyme led to the identification of over 100 distinct alkane monooxygenase (AlkB) enzymes containing a covalently bound, or fused, rubredoxin. The rubredoxin-fused AlkB from Dietzia cinnamea was cloned as a f...
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Veröffentlicht in: | Journal of inorganic biochemistry 2021-06, Vol.219, p.111409-111409, Article 111409 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | Interest in understanding the environmental distribution of the alkane monooxygenase (AlkB) enzyme led to the identification of over 100 distinct alkane monooxygenase (AlkB) enzymes containing a covalently bound, or fused, rubredoxin. The rubredoxin-fused AlkB from Dietzia cinnamea was cloned as a full-length protein and as a truncated protein with the rubredoxin domain deleted. A point mutation (V91W) was introduced into the full-length protein, with the goal of assessing how steric bulk in the putative substrate channel might affect selectivity. Based on activity studies with alkane and alkene substrates, the rubredoxin-fused AlkB oxidizes a similar range of alkane substrates relative to its rubredoxin domain-deletion counterpart. Oxidation of terminal alkenes generated both an epoxide and a terminal aldehyde. The products of V91W-mutant-catalyzed oxidation of alkenes had a higher aldehyde-to-epoxide ratio than the products formed in the presence of the wild type protein. These results are consistent with this mutation causing a structural change impacting substrate positioning.
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•Alkane monooxygenases with fused rubredoxin widely distributed in nature•Rubredoxin deletion mutant has similar substrate range to wild type•Single point mutation leads to an enzyme with substantial anti-Markovnikov activity on alkenes |
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ISSN: | 0162-0134 1873-3344 |
DOI: | 10.1016/j.jinorgbio.2021.111409 |