Liu Qin, Ma Yue, Zhou Lingyun, Zhang Yuanxing
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, China.
Arch Microbiol. 2006 Mar;185(2):159-63. doi: 10.1007/s00203-005-0058-4. Epub 2006 Jan 21.
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).
酰基载体蛋白（ACP）是一种小的酸性蛋白，在许多依赖酰基转移反应的生物合成途径中作为必需的辅助因子发挥作用。在本研究中，首先从强毒鳗弧菌菌株MVM425的染色体中克隆了鳗弧菌ACP编码基因acpV。acpV在大肠杆菌中过表达，并纯化得到了相应的蛋白AcpV。将纯化的AcpV与纯化的磷酸泛酰巯基乙胺基转移酶（PPtase）在辅酶A存在的情况下孵育，以在体外检测AcpV的4'-磷酸泛酰巯基乙胺化；另一方面，将acpV基因与PPtase编码基因在大肠杆菌中共表达，以在体内检测AcpV的4'-磷酸泛酰巯基乙胺化。我们的结果表明，acpV编码一种鳗弧菌功能性ACP，其在体外和体内均可被AcpS型PPtase（大肠杆菌AcpS）很好地进行4'-磷酸泛酰巯基乙胺化，但不能作为Sfp型PPtase（鳗弧菌AngD）的良好底物。
