Production of A Stable Recombinant α-amylase Enzyme from Thermoanaerobacterium: Thermoanaerobacterium in Pilot Scale Fermenter

Abstract

This study investigated the production of stable α-amylase from thermopile bacteria Thermoanaerobacterium thermoanaerobacterium . The gene encoding this enzyme was cloned and expressed in E. coli using IPTG as the standard inducer of T7 promoter. For large-scale production optimization, several parameters were adjusted. High recombinant enzyme yield was achieved u nde r conditions where the medium volume comprised 60% of the fermenter’s total volume, aeration rate was set at 2.5 vvm, dissolved oxygen was maintained at 15%, and agitation speed of 150 rpm was used. Using the optimized conditions, highest enzyme production was reached to 6.17 ± 0.32 U/ml/min with total proteins of 4.73 ± 0.04 mg/ml and viable cells 5.82 ×10 7 cells/ml. When the E. coli cells were induced with IPTG and lactose, maximum enzyme production (16.8 U/ml/min) was obtained when induction was done with 0