Production of therapeutically relevant indolizidine alkaloids in Securinega suffruticosa in vitro shoots maintained in liquid culture systems.

Abstract

Microshoot cultures of the Chinese medicinal plant Securinega suffruticosa (Pall.) Rehd. were established and evaluated for the presence of therapeutically relevant indolizidine alkaloids securinine (S) and allosecurinine (AS). The cultures were maintained in shake flasks (SFs) and a bubble column bioreactor (BCB) using the modified Murashige’s shoot multiplication medium supplemented with 1.0 mg l−1 benzyladenine (BA), 3.0 mg l−1 2-isopentenyladenine (2iP), and 0.3 mg l−1 1-naphthaleneacetic acid (NAA). The influence of light and medium supplementation strategies with biosynthesis precursor (lysine (LY)) and nutrient formulations (casein hydrolysate (CH) and coconut water (CW)) on biomass growth and alkaloid production were investigated. SF cultures grown in the presence of light yielded up to 6.02 mg g−1 dry weight (DW) S and 3.70 mg g−1 DW AS, corresponding to the respective productivities of 98.39 and 60.21 mg l−1. Among feeding experiments, CW supplementation proved most effective for SF-grown shoots, increasing biomass yield and AS productivity by 52 and 44 %, respectively. Maximum concentrations of securinine (3.25 mg g−1 DW) and allosecurinine (3.41 mg g−1 DW) in BCB cultures were achieved in the case of 1.0 g l−1 LY supplementation. These values corresponded to the productivities of 42.64 and 44.47 mg per bioreactor, respectively.