Abstract
Acyl carrier protein (ACP) is a small acidic protein that acts as an essential cofactor in many biosynthetic pathways depending on acyl transfer reactions. In this work, a Vibrio anguillarum ACP encoding gene, acpV, was first cloned from the chromosome of a virulent V. anguillarum strain MVM425. acpV was over-expressed in Escherichia coli and the resultant protein AcpV was purified. The purified AcpV was incubated with purified phosphopantetheinyl transferase (PPtase) in the presence of CoA to assay the 4′-phosphopantetheinylation of AcpV in vitro; and on the other hand, the acpV gene was co-expressed with PPtase-encoding gene in E. coli to examine the 4′-phosphopantetheinylation of AcpV in vivo. Our results suggested that acpV encoded a functional ACP of V. anguillarum, which can be 4′-phosphopantetheinylated well by AcpS-type PPtase (E. coli AcpS) both in vitro and in vivo, but cannot serve as a good substrate for Sfp-type PPtase (V. anguillarum AngD).
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Acknowledgements
We especially thank Dr. Christopher T. Walsh (the Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, USA) for the gift of recombinant plasmid pDPJ. This work was supported by the grants from the National High-Tech Research and Development Programs of China (no. 2003AA622020).
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Liu, Q., Ma, Y., Zhou, L. et al. Gene cloning, expression and functional characterization of an acyl carrier protein AcpV from Vibrio anguillarum . Arch Microbiol 185, 159–163 (2006). https://doi.org/10.1007/s00203-005-0058-4
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DOI: https://doi.org/10.1007/s00203-005-0058-4