A thin layer chromatographic comparison of raw and soluble starch hydrolysis patterns of some α-amylases from Bacillus sp. isolated in Serbia

Abstract

Several natural isolates of Bacillus strains namely 5B, 12B, 16B, 18 and 24B were grown at two different temperatures in submerged fermentation for the production of raw-starch-digesting α-amylases. All strains except Bacillus sp. 18 produced more α-amylase at 37 °C. The hydrolysis of raw cornstarch followed the same pattern. Efficient hydrolysis was obtained with α-amylases from Bacillus sp. 5B, 12B, 16B and 24B grown at 37 °C and Bacillus sp. 18 grown at 50 °C. Zymography after isoelectric focusing showed that α-amylases were produced in multiple forms, from 2 to 6, depending on the strain when they were growing at 37 °C, while growth at 50 °C induced only 1 or 2 isoforms. Thin layer chromatography (TLC) analysis of the hydrolysis products of raw corn and soluble starch by α-amylases revealed the production of various mixtures of oligosaccharides. In most cases, G3 was the most dominant product from soluble starch while G2, G3 and G5 were the main products of raw starch hydrolysis. This indicates that the obtained α-amylases could be used for starch liquidification or short-chain-oligosaccharide formation, depending on the type of starch (raw or soluble) used for the hydrolysis.

Characterization of Two Purified Amylase Produced from <i>Aspergillus niger</i> ICP2 and its Immobilization Using Activated Carbon

“…Visualization was employed using a solution containing 2.5 ml H2SO4 and 47.5 absolute ethanol [24]. The plate was dried using a dryer and then placed in an oven at 110°C for 10 minutes to visualize the sugar contained [25].…”