Abstract
A Bacillus sp. TS-23 α-amylase produced by recombinant Escherichia coli was adsorbed onto raw starch and the adsorbed enzyme was eluted with maltose or maltodextrin in 50 mM Tris/HCI buffer (pH 8.5). The adsorption-elution procedure resulted in a yield of 53% α-amylase activity and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS/PAGE) analysis showed that the eluted α-amylase had a molecular mass of approximately 64 kDa. Raw starch could be used repeatedly in the adsorption-elution cycle with good reproducibility. Scanning electron microscopy of the isolated corn starch exhibited a smooth appearance of the granules before adsorption and only a small change in appearance after three adsorption-elution cycles. These results suggest that the raw starch adsorption-elution technique has a great potential in the isolation of Bacillus sp. TS-23 α-amylase from the culture broth of recombinant E. coli.
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