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KMID : 1094720110160050894
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Biotechnology and Bioprocess Engineering
2011 Volume.16 No. 5 p.894 ~ p.894
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A new microtiter plate-based screening method for microorganisms producing Alpha-amylase inhibitors
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Feng Zhi-Hua
Wang Yuan-Shan
Zheng Yu-Guo
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Abstract
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Alpha-amylase inhibitors are widely used by the pharmaceutical and agricultural industries, such as the treatment of diabetes and obesity and insect controller. Here, we developed a colorimetric method to screen for α-amylase inhibitor producing strains or mutants with higher α-amylase inhibitor productivity. This method relies on absorbance changes at 402 nm that are due to the inhibition of α-amylase catalyzed hydrolysis of 2-Chloro-4-nitrophenyl-4-O-β-D-galactopyranosyl-maltoside by α-amylase inhibitors. The assay can be performed on a microtiter plate, making it simple and convenient. Using this method, α-amylase inhibitor producing strains and mutants with higher α-amylase inhibitor productivity can be rapidly screened. One strain, ZJB-08196, with the highest α-amylase inhibition was isolated and identified as Actinoplanes utahensis, and one mutant with higher acarbose production was obtained by screening 3,000 variants using this method.
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KEYWORD
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α-amylase inhibitors, human salivary amylase, 2-Chloro-4-nitrophenyl-4-O-β-D-galactopyrano-syl-maltoside, diabetes mellitus, acarbose
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