Gene cloning, expression and functional characterization of an acyl carrier protein AcpV from Vibrio anguillarum.
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).
Liu
Qin
Q
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, China.
Ma
Yue
Y
Zhou
Lingyun
L
Zhang
Yuanxing
Y
D000213:Acyl Carrier Protein / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D010204:Pantetheine / Q000031:analogs & derivatives; D017855:Transferases (Other Substituted Phosphate Groups) / Q000378:metabolism; D014733:Vibrio / Q000235:genetics* / Q000378:metabolism
D000213:Acyl Carrier Protein; D001426:Bacterial Proteins; C082662:phosphopantetheinyl transferase; D010204:Pantetheine; D017855:Transferases (Other Substituted Phosphate Groups); C003129:4'-phosphopantetheine