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molecular cloning and characterization of three phenylalanine ammonia-lyase genes from Schisandra chinensis

Chinese Journal of Natural Medicines 2022年第7期20卷 527-536页

作者： FAN San-Peng CHEN Wei WEI Jiang-Chun GAO Xiao-Xu YANG Yong-Cheng WANG An-Hua HU Gao-Sheng JIA Jing-Ming School of Traditional Chinese Materia Medica Shenyang Pharmaceutical UniversityShenyang 110016China

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 pathway of plants and *** chinensis,a woody vine plant belonging to the family of Magnoliaceae,is a rich source of dibenzocyclooctadiene lignans exhibiting potent ***,the functional role of PAL in the biosynthesis of lignan is relatively limited,compared with those in lignin and flavonoids ***,it is essential to clone and characterize the PAL genes from this valuable medicinal *** this study,molecular cloning and characterization of three PAL genes(ScPAL1−3)from *** was carried *** were cloned using RACE *** sequence analysis of the three ScPALs was carried out to give basic characteristics followed by docking *** 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 *** catalytic product of the purified recombinant proteins was verified using RP-HPLC through comparing with standard *** optimal temperature,pH value and effects of different metal ions were ***,Kcat and Km values were determined under the optimal *** expression of three ScPALs in different tissues was also *** work provided essential information for the function of ScPALs.

关键词： Schisandra chinensis Phenylalanine ammonia-lyase molecular cloning and characterization Kinetics

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