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

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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 under 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 ×107 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 0.5 mM lactose. Maximum protein produced was 9.62 ± 0.24 mg/ml.