TY - JOUR
T1 - Structure-Based Engineering of Strictosidine Synthase
T2 - Auxiliary for Alkaloid Libraries
AU - Loris, Elke A.
AU - Panjikar, Santosh
AU - Ruppert, Martin
AU - Barleben, Leif
AU - Unger, Matthias
AU - Schübel, Helmut
AU - Stöckigt, Joachim
PY - 2007/9/21
Y1 - 2007/9/21
N2 - 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.
AB - 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.
KW - CHEMBIOL
KW - MICROBIO
UR - http://www.scopus.com/inward/record.url?scp=34548694786&partnerID=8YFLogxK
U2 - 10.1016/j.chembiol.2007.08.009
DO - 10.1016/j.chembiol.2007.08.009
M3 - Article
C2 - 17884630
AN - SCOPUS:34548694786
VL - 14
SP - 979
EP - 985
JO - Chemistry and Biology
JF - Chemistry and Biology
SN - 1074-5521
IS - 9
ER -