Cloning, prokaryotic expression and functional analysis of squalene synthase (SQS) in Magnolia officinalis

摘要

Magnolia officinalis Rehder et Wilson is a traditional Chinese herbal medicine that is used to treat various diseases such as neurosis, anxiety, and stroke. The main secondary metabolites in magnolia bark are phenolic compounds and terpenoids. Squalene synthase plays a significant role in catalyzing two farnesyl diphosphate molecules to form squalene, the first precursor of triterpenoid, phytosterol, and cholesterol biosynthesis. In this study, a full-length cDNA of squalene synthase was cloned from M. officinalis and designated MoSQS (GenBank accession no. KT223496). The gene contains a 1240-bp open reading frame and it encodes a protein with 409 amino acids. Bioinformatic and phylogenetic analysis clearly suggested that MoSQS shared high similarity with squalene synthases among other plants. Prokaryotic expression showed that a transmembrane domain-deleted (385-409 aa) MoSQS mutant (MoSQSA Delta TM) could be expressed in its soluble form in Escherichia coli Transetta (DE3). GC MS analysis showed that squalene was detected in an in vitro reaction mixture. These results indicated that MoSQS Delta TM was functional, thereby establishing an important foundation for the study of triterpenoid biosynthesis in M. officinalis. (C) 2015 Elsevier Inc. All rights reserved.