Premium
Improved Expression of His 6 ‐Tagged Strictosidine Synthase cDNA for Chemo‐Enzymatic Alkaloid Diversification
Author(s) -
Yang Liuqing,
Zou Hongbin,
Zhu Huajian,
Ruppert Martin,
Gong Jingxu,
Stöckigt Joachim
Publication year - 2010
Publication title -
chemistry and biodiversity
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.427
H-Index - 70
eISSN - 1612-1880
pISSN - 1612-1872
DOI - 10.1002/cbdv.201000052
Subject(s) - chemistry , tryptamine , enzyme , biochemistry , biosynthesis , indole test , escherichia coli , stereochemistry , gene
Strictosidine synthase (STR1) catalyzes the stereoselective formation of 3 α ( S )‐strictosidine from tryptamine and secologanin. Strictosidine is the key intermediate in the biosynthesis of 2,000 plant monoterpenoid indole alkaloids, and it is a key precursor of enzyme‐mediated synthesis of alkaloids. An improved expression system is described which leads to optimized His 6 ‐STR1 synthesis in Escherichia coli. Optimal production of STR1 was achieved by determining the impact of co‐expression of chaperones pG‐Tf2 and pG‐LJE8. The amount and activity of STR1 was doubled in the presence of chaperone pG‐Tf2 alone. His 6 ‐STR1 immobilized on Ni‐NTA can be used for enzymatic synthesis of strictosidines on a preparative scale. With the newly co‐expressed His 6 ‐STR1, novel 3 α ( S )‐12‐azastrictosidine was obtained by enzymatic catalysis of 7‐azatryptamine and secologanin. The results obtained are of significant importance for application to chemo‐enzymatic approaches leading to diversification of alkaloids with novel improved structures.