姜黄倍半萜合酶基因ClFLDH的克隆和原核表达分析OA

The farnesol dehydrogenase gene(ClFLDH)of Curcuma longa is a key rate-limiting enzyme in the terpene compound synthesis pathway.To explore the improvement of terpene compound production in C.longa,the cDNA sequence of ClFLDH was obtained through homologous cloning.The open reading frame(ORF)of ClFLDH was 855 bp,encoding 295 amino acids.Bioinformatics analysis indicated that the relative molecular mass of ClFLDH protein was 31 kDa,with an isoelectric point of 6.79,and it was a liposoluble,hydrophobic and neutral protein.The pCE32a-FLDH prokaryotic expression vector was constructed and expressed in Escherichia coli DH5α.The SDS-PAGE electrophoresis results of the recombinant strain showed that the size of the target protein was 37 kDa.GC-MS analysis indicated that the ClFLDH protein might catalyze 3,5-dimethoxyphenol to generate[1,2-a]pyrazine-1,4-dione,6H-3-(benzyl)-pyrrole.These would provide a new perspective for in-depth understanding of the regulatory mechanism of curcuminoid terpene synthesis and lay a foundation for increasing the production of curcuminoid terpene compounds through genetic engineering.