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

Paper ID Volume ID Publish Year Pages File Format Full-Text
20865 43195 2013 5 PDF Available
Title
Biosynthesis of a stable allophycocyanin beta subunit in metabolically engineered Escherichia coli
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.

Keywords
Allophycocyanin; Biosynthesis; Escherichia coli; Fluorescence; Thermostability
First Page Preview
Biosynthesis of a stable allophycocyanin beta subunit in metabolically engineered Escherichia coli
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 115, Issue 5, May 2013, Pages 485–489
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering