Structure-based engineering of strictosidine synthase: auxiliary for alkaloid libraries.
The highly substrate-specific strictosidine synthase (EC 4.3.3.2) catalyzes the biological Pictet-Spengler condensation between tryptamine and secologanin, leading to the synthesis of about 2000 monoterpenoid indole alkaloids in higher plants. The crystal structure of Rauvolfia serpentina strictosidine synthase (STR1) in complex with strictosidine has been elucidated here, allowing the rational site-directed mutation of the active center of STR1 and resulting in modulation of its substrate acceptance. Here, we report on the rational redesign of STR1 by generation of a Val208Ala mutant, further describing the influence on substrate acceptance and the enzyme-catalyzed synthesis of 10-methyl- and 10-methoxystrictosidines. Based on the addition of strictosidine to a crude strictosidine glucosidase preparation from Catharanthus cells, a combined chemoenzymatic approach to generating large alkaloid libraries for future pharmacological screenings is presented.
Loris
Elke A
EA
Department of Pharmaceutical Biology, Institute of Pharmacy, Johannes Gutenberg-University Mainz, Staudingerweg 5, D-55099 Mainz, Germany.
Panjikar
Santosh
S
Ruppert
Martin
M
Barleben
Leif
L
Unger
Matthias
M
Schübel
Helmut
H
Stöckigt
Joachim
J
D019759:Carbon-Nitrogen Lyases / Q000737:chemistry* / Q000235:genetics*; D018360:Crystallography, X-Ray; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D026121:Indole Alkaloids / Q000737:chemistry*; D015394:Molecular Structure; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D010940:Plant Proteins; D011485:Protein Binding / Q000235:genetics; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D013379:Substrate Specificity / Q000235:genetics; D014748:Vinca Alkaloids / Q000737:chemistry*
D026121:Indole Alkaloids; D010940:Plant Proteins; D014748:Vinca Alkaloids; C021093:strictosidine; D019759:Carbon-Nitrogen Lyases; C022684:strictosidine synthetase