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J Biosci Bioeng. 2013 May;115(5):485-9. doi: 10.1016/j.jbiosc.2012.11.008. Epub 2012 Dec 21.

Biosynthesis of a stable allophycocyanin beta subunit in metabolically engineered Escherichia coli.

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  • 1Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.

Abstract

Allophycocyanin (APC) is widely used as a fluorescent tag for fluorescence detection techniques. In this study, the apcB gene from a thermophilic cyanobacterium strain was cloned and ligated into an expression vector to construct a pathway for the biosynthesis of an allophycocyanin beta subunit (holo-apcBT) in Escherichia coli. Isopropyl β-d-1-thiogalactopyranoside induction successfully reconstituted holo-apcBT in E. coli. The recombinant holo-apcB showed spectroscopic properties similar to native APC. The stability and spectroscopic properties of this protein were then compared with another recombinant allophycocyanin beta subunit (holo-apcBM) whose apcB gene was cloned from mesophilic cyanobacterium. At high temperatures and during the course of storage, holo-apcBT was significantly more stable than holo-apcBM. In addition, holo-apcBT had an unexpectedly higher extinction coefficient and fluorescence quantum yield than holo-apcBM, suggesting that holo-apcBT would be a promising fluorescent tag and serve as a substitute for native APC.

Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

[PubMed - indexed for MEDLINE]
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