典型文献
Molecular cloning and characterization of three phenylalanine ammonia-lyase genes from Schisandra chinensis
文献摘要:
Phenylalanine ammonia-lyase(PAL),which catalyzes the conversion from L-phenylalanine to trans-cinnamic acid,is a well-known key enzyme and a connecting step between primary and secondary metabolisms in the phenylpropanoid biosynthetic path-way of plants and microbes.Schisandra chinensis,a woody vine plant belonging to the family of Magnoliaceae,is a rich source of dibenzocyclooctadiene lignans exhibiting potent activity.However,the functional role of PAL in the biosynthesis of lignan is relat-ively limited,compared with those in lignin and flavonoids biosynthesis.Therefore,it is essential to clone and characterize the PAL genes from this valuable medicinal plant.In this study,molecular cloning and characterization of three PAL genes(ScPAL1-3)from S.chinensis was carried out.ScPALs were cloned using RACE PCR.The sequence analysis of the three ScPALs was carried out to give basic characteristics followed by docking analysis.In order to determine their catalytic activity,recombinant protein was obtained by heterologous expression in pCold-TF vector in Escherichia coli(BL21-DE3),followed by Ni-affinity purification.The catalytic product of the purified recombinant proteins was verified using RP-HPLC through comparing with standard compounds.The optimal temperature,pH value and effects of different metal ions were determined.Vmax,Kcatand Km values were determined under the optim-al conditions.The expression of three ScPALs in different tissues was also determined.Our work provided essential information for the function of ScPALs.
文献关键词:
中图分类号:
作者姓名:
FAN San-Peng;CHEN Wei;WEI Jiang-Chun;GAO Xiao-Xu;YANG Yong-Cheng;WANG An-Hua;HU Gao-Sheng;JIA Jing-Ming
作者机构:
School of Traditional Chinese Materia Medica,Shenyang Pharmaceutical University,Shenyang 110016,China
文献出处:
引用格式:
[1]FAN San-Peng;CHEN Wei;WEI Jiang-Chun;GAO Xiao-Xu;YANG Yong-Cheng;WANG An-Hua;HU Gao-Sheng;JIA Jing-Ming-.Molecular cloning and characterization of three phenylalanine ammonia-lyase genes from Schisandra chinensis)[J].中国天然药物,2022(07):527-536
A类:
dibenzocyclooctadiene,ScPAL1,ScPALs,Kcatand
B类:
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AB值:
0.573275
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