A Maltotriose producing thermostable amylase from Bacillus sp KR11

Moumita Karmakar, Rina Rani Ray

Abstract


A bacterial strain of Bacillus sp KR 11 was isolated from soil which was found to produce extra cellular maltotriose producing amylase. The enzyme was partially purified from crude culture filtrate by ammonium sulfate fractionation, followed by dialysis. The optimum pH and temperature for the purified enzyme were 7.0 and 65 °C, respectively. The thermo inactivation kinetics indicated that the half life of the enzyme at 65°C was 60 minutes. The enzyme activity was completely inhibited by 10 mM Hg2+ and Cu2+ and thiol inhibitor like pCMB. Restoration of enzyme activity in presence of external thiols like cysteine, reduced glutathione and dithiothreitol indicated the presence of thiol groups at the active site. The starchy residues collected from various wastes could be converted into sugars, of which starch extracted from tamarind kernel showed the highest yield of sugar. Thin layer chromatographic analysis indicated the presence of maltotriose as the major end product of amylase action of these starches. The enzyme immobilized on 7% gelatin powder crosslinked with glutaraldehyde showed the best result and could be used for judicious exploitation of this enzyme in future.

 

Key words: amylase, Bacillus sp, maltotriose, saccharification, immobilization.


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