title: Identification and characterization of L-lysine decarboxylase from Huperzia serrata and its role in the metabolic pathway of lycopodium alkaloid.
abstract: Lysine decarboxylation is the first biosynthetic step of Huperzine A (HupA). Six cDNAs encoding lysine decarboxylases (LDCs) were cloned from Huperzia serrata by degenerate PCR and rapid amplification of cDNA ends (RACE). One HsLDC isoform was functionally characterized as lysine decarboxylase. The HsLDC exhibited greatest catalytic efficiency (kcat/Km, 2.11 s-1mM-1) toward L-lysine in vitro among all reported plant-LDCs. Moreover, transient expression of the HsLDC in tobacco leaves specifically increased cadaverine content from zero to 0.75 mg per gram of dry mass. Additionally, a convenient and reliable method used to detect the two catalytic products was developed. With the novel method, the enzymatic products of HsLDC and HsCAO, namely cadaverine and 5-aminopentanal, respectively, were detected simultaneously both in assay with purified enzymes and in transgenic tobacco leaves. This work not only provides direct evidence of the first two-step in biosynthetic pathway of HupA in Huperzia serrata and paves the way for further elucidation of the pathway, but also enables engineering heterologous production of HupA.
authors.affiliation: CAS Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China; University of Chinese Academy of Sciences, Beijing, 100039, China.
authors.forename: Baofu
authors.initials: B
authors.lastname: Xu
authors.affiliation: CAS Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Lei
authors.affiliation: CAS Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China.
authors.forename: Xiaocen
authors.initials: X
authors.lastname: Zhu
authors.affiliation: CAS Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China.
authors.forename: Yiqing
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: CAS Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China; University of Chinese Academy of Sciences, Beijing, 100039, China; CAS-JIC Centre of Excellence in Plant and Microbial Sciences, China. Electronic address: ylxiao@sibs.ac.cn.
authors.forename: Youli
authors.initials: Y
authors.lastname: Xiao
mesh_terms: D000470:Alkaloids / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D006631:Amine Oxidase (Copper-Containing) / Q000378:metabolism*; D002103:Cadaverine / Q000032:analysis; D002262:Carboxy-Lyases / Q000378:metabolism* / Q000493:pharmacokinetics; D018076:DNA, Complementary / Q000378:metabolism; D032501:Huperzia / Q000201:enzymology*; D032502:Lycopodium / Q000737:chemistry*; D008239:Lysine / Q000378:metabolism; D053858:Metabolic Networks and Pathways; D015394:Molecular Structure; D018515:Plant Leaves / Q000737:chemistry; D030821:Plants, Genetically Modified / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012717:Sesquiterpenes / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000470:Alkaloids; D018076:DNA, Complementary; D012717:Sesquiterpenes; C050426:huperzine A; D006631:Amine Oxidase (Copper-Containing); D002262:Carboxy-Lyases; C021520:lysine decarboxylase; D008239:Lysine; D002103:Cadaverine
keywords: 5-Aminopentanal; Copper amine oxidase; Huperzia serrata; Huperzine A; L-lysine decarboxylase; Lycopodiaceae