Enhanced production of napthoquinone metabolite (shikonin) from cell suspension culture of Arnebia sp. and its up-scaling through bioreactor.

Abstract

Cell culture in shake flask and air-lift bioreactor was carried out to exploit the potential of Arnebia sp. for napthoquinone metabolite production. Cell suspension cultures of Arnebia were established from friable callus in liquid MS medium supplemented with 6-benzylaminopurine (BAP) (10 μM) and indole-3-butyric acid (IBA) (5 μM). Growth kinetic studies were done by using settled cell volume and fresh/dry cell weight method. Suspension cultures were maintained by sub-culturing at 10 days interval. A two-stage culture system is employed using growth medium (GM) and modified M9 medium (production medium) for cell biomass and naphthoquinone pigment production, respectively. Results showed that cultivation of cells under dark conditions at room temperature (25 ± 2 °C) enhanced the cell biomass from 100 to 625 g l−1. The pigment production was also found to be increased in dark conditions at room temperature. Alkaline pH found to have positive effect on pigment yield. In case of M9 medium constituents, absence of Na2SO4 does not affect the pigment yield. The current approaches have the cumulative effect to meet an increased level of (25.5 μg/ml) metabolite production in air-lift bioreactor.