6533b837fe1ef96bd12a1d25

RESEARCH PRODUCT

Structure-based engineering of strictosidine synthase: auxiliary for alkaloid libraries.

Elke A. LorisHelmut SchübelSantosh PanjikarLeif BarlebenMatthias UngerJoachim StöckigtJoachim StöckigtMartin Ruppert

subject

TryptamineCHEMBIOLStrictosidine synthaseMICROBIOStereochemistryProtein ConformationClinical BiochemistryMutantDrug Evaluation PreclinicalMutation MissenseCrystallography X-RayProtein EngineeringBiochemistryIndole AlkaloidsSubstrate Specificitychemistry.chemical_compoundRauvolfia serpentinaDrug DiscoveryCatharanthusCarbon-Nitrogen LyasesMolecular BiologyVinca AlkaloidsPlant ProteinsPharmacologybiologyMolecular StructureGeneral Medicinebiology.organism_classificationLyaseBiochemistrychemistryStrictosidinebiology.proteinMutagenesis Site-DirectedMolecular MedicineSecologaninProtein Binding

description

SummaryThe 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.

yearjournalcountryeditionlanguage
2007-09-01Chemistrybiology
10.1016/j.chembiol.2007.08.009https://pubmed.ncbi.nlm.nih.gov/17884630