6533b7cefe1ef96bd1257063

RESEARCH PRODUCT

Construction and expression of a dual vector for chemo-enzymatic synthesis of plant indole alkaloids inEscherichia coli

Bodo HammesMartin RuppertJoachim Stöckigt

subject

TryptamineDNA ComplementaryStrictosidine synthasePlant Sciencemedicine.disease_causeBiochemistryGene Expression Regulation EnzymologicRauwolfiaIndole AlkaloidsAnalytical Chemistrychemistry.chemical_compoundGene Expression Regulation PlantRauvolfia serpentinaCarbon-Nitrogen LyasesEscherichia colimedicineCloning MolecularEscherichia coliPlant ProteinsIndole testchemistry.chemical_classificationMolecular StructurebiologyIndole alkaloidOrganic Chemistrybiology.organism_classificationSecologanin Tryptamine AlkaloidsEnzymechemistryBiochemistrybiology.proteinSecologaninGlucosidases

description

A dual vector (pQE-70-STR1-SG) containing coding regions of strictosidine synthase (STR1, EC 4.3.3.2) and strictosidine glucosidase (SG, EC 3.2.1.105) from the Indian medicinal plant Rauvolfia serpentina was constructed. Functional expression of the vector in Escherichia coli cells (M15 strain) was proven by isolation of prepurified enzyme extracts, which show both STR1 and SG activities. Incubation of the enzyme in the presence of tryptamine and secologanin delivered the indole alkaloid cathenamine, demonstrating functional co-expression of both STR1- and SG-cDNAs. Cathenamine reduction by sodium borohydride leading to tetrahydroalstonine revealed the chemo-enzymatic indole alkaloid synthesis.

https://doi.org/10.1080/14786410903247304