Department of Bioresources, New Drug R&D Center of North China Pharmaceutical Corporation and National Engineering Research Center for Microbial Medicine, 388 East Heping Road, Shijiazhuang, 050015, China.
Folia Microbiol (Praha). 2011 May;56(3):246-52. doi: 10.1007/s12223-011-0044-y. Epub 2011 May 31.
A novel phenylacetic acid (PAA)-induced CoA-ligase-encoding gene, designated as phlC, has been cloned from penicillin-producing fungus Penicillium chrysogenum. The open reading frame of phlC cDNA was 1671 bp and encoded a 556 amino acid residues protein with the consensus AMP binding site and a peroxisomal targeting signal 1 on its C terminus. The deduced amino acid sequence showed 37% and 38% identity with characterized P. chrysogenum Phl and PhlB protein, respectively. Functional recombinant PhlC protein was overexpressed in Escherichia coli. The purified recombinant enzyme was capable to convert PAA into its corresponding CoA ester with a specific activity of 129.5 ± 3.026 pmol/min per mg protein. Similar to Phl and PhlB, PhlC displayed broad substrate spectrum and showed higher activities to medium- and long-chain fatty acids. The catalytic properties of PhlC have been determined and compared to those of Phl and PhlB.
从青霉素产生菌产黄青霉中克隆到一个新型苯乙酸（PAA）诱导的辅酶 A 连接酶编码基因，命名为 phlC。phlC cDNA 的开放阅读框为 1671bp，编码一个 556 个氨基酸残基的蛋白质，其 C 末端具有 AMP 结合位点和过氧化物酶体靶向信号 1。推导的氨基酸序列与已鉴定的产黄青霉 Phl 和 PhlB 蛋白分别具有 37%和 38%的同一性。功能重组 PhlC 蛋白在大肠杆菌中过表达。纯化的重组酶能够将 PAA 转化为相应的 CoA 酯，比活为 129.5±3.026pmol/min per mg 蛋白。与 Phl 和 PhlB 相似，PhlC 显示出广泛的底物谱，对中链和长链脂肪酸表现出更高的活性。已经测定了 PhlC 的催化特性，并与 Phl 和 PhlB 进行了比较。
