title: Cloning and functional characterization of a 4-coumarate CoA ligase from liverwort Plagiochasma appendiculatum.
abstract: Plant phenylpropanoids represent a large group of secondary metabolites which have played an important role in terrestrial plant life, beginning with the evolution of land plants from primitive green algae. 4-Coumarate: coenzyme A ligase (4CL) is a provider of activated thioester substrates within the phenylpropanoid synthesis pathway. Although 4CLs have been extensively characterized in angiosperm, gymnosperm and moss species, little is known of their functions in liverworts. Here, a 4CL homolog (designated as Pa4CL1) was isolated from the liverwort species Plagiochasma appendiculatum. The full-length cDNA sequence of Pa4CL1 contains 1644bp and is predicted to encode a protein with 547amino acids. The gene products were 40-50% identical with 4CL sequences reported in public databases. The recombinant protein was heterologously expressed in Escherichia coli and exhibited a high level of 4CL activity, catalyzing formation of hydroxycinnamate-CoA thioesters by a two-step reaction mechanism from corresponding hydroxycinnamic acids. Kinetic analysis indicated that the most favorable substrate for Pa4CL1 is p-coumaric acid. The transcription of Pa4CL1 was induced when P. appendiculatum thallus was treated with either salicylic acid or methyl jasmonate.
authors.affiliation: Key Laboratory of Chemical Biology of Natural Products, Ministry of Education, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Gao
authors.affiliation: Key Laboratory of Chemical Biology of Natural Products, Ministry of Education, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Hai-Na
authors.initials: HN
authors.lastname: Yu
authors.affiliation: Key Laboratory of Chemical Biology of Natural Products, Ministry of Education, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Rui-Xue
authors.initials: RX
authors.lastname: Xu
authors.affiliation: Key Laboratory of Chemical Biology of Natural Products, Ministry of Education, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China. Electronic address: aixiacheng@sdu.edu.cn.
authors.forename: Ai-Xia
authors.initials: AX
authors.lastname: Cheng
authors.affiliation: Key Laboratory of Chemical Biology of Natural Products, Ministry of Education, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China. Electronic address: louhongxiang@sdu.edu.cn.
authors.forename: Hong-Xiang
authors.initials: HX
authors.lastname: Lou
mesh_terms: D000085:Acetates / Q000737:chemistry / Q000378:metabolism; D001632:Bibenzyls / Q000737:chemistry / Q000378:metabolism; D019976:Cloning, Organism; D003066:Coenzyme A Ligases / Q000235:genetics / Q000378:metabolism*; D003373:Coumaric Acids / Q000737:chemistry / Q000378:metabolism; D003517:Cyclopentanes / Q000737:chemistry / Q000378:metabolism; D005419:Flavonoids / Q000737:chemistry / Q000378:metabolism; D035926:Hepatophyta / Q000737:chemistry* / Q000201:enzymology / Q000235:genetics; D007700:Kinetics; D008969:Molecular Sequence Data; D054883:Oxylipins / Q000737:chemistry / Q000378:metabolism; D010666:Phenylpropionates / Q000737:chemistry / Q000378:metabolism; D011422:Propionates; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D000085:Acetates; D001632:Bibenzyls; D003373:Coumaric Acids; D003517:Cyclopentanes; D005419:Flavonoids; D054883:Oxylipins; D010666:Phenylpropionates; D011422:Propionates; D011994:Recombinant Proteins; C072239:methyl jasmonate; D003066:Coenzyme A Ligases; C019943:4-coumarate-CoA ligase; C495469:p-coumaric acid
keywords: 4-Coumarate: coenzyme A ligase; Aytoniaceae; Bis-bibenzyls; Flavonoids; Liverwort; Marchantiales; Phenylpropanoids; Plagiochasma appendiculatum