An analytical method for measuring α-amylase activity in starch-containing foods

• Koyama, Kazuo

• Hirao, Takashi

• Toriba, Akira

• Hayakawa, Kazuichi

抄録

The quality of starch-containing foods may be significantly impaired by contamination with very small amounts of α-amylase, which can enzymatically hydrolyze the starch and cause viscosity loss. Thus, for quality control, it is necessary to have an analytical method that can measure low amylase activity. We developed a sensitive analytical method for measuring the activity of α-amylase (from Bacillus subtilis) in starch-containing foods. The method consists of six steps: (1) crude extraction of α-amylase by centrifugation and filtration; (2) α-amylase purification by desalting and anion-exchange chromatography; (3) reaction of the purified amylase with boron-dipyrromethene (BODIPY)-labeled substrate, which releases a fluorescent fragment upon digestion of the substrate, thus avoiding interference from starch derivatives in the sample; (4) stopping the reaction with acetonitrile; (5) reversed-phase solid-phase extraction of the fluorescent substrate to remove contaminating dye and impurities; and (6) separation and measurement of BODIPY fluorescence by HPLC. The proposed method could quantify α-amylase activities as low as 10mU/mL, which is enough to reduce the viscosity of starch-containing foods. © 2012 John Wiley & Sons, Ltd.

収録刊行物

• Biomedical Chromatography

  Biomedical Chromatography 27 (5), 583-588, 2013-05-01

  Wiley-Blackwell

キーワード

• α-amylase
• Fluorescent substrate
• High-performance liquid chromatography
• Purification
• Starch-containing foods

詳細情報

• CRID: 1050845760884115328
• NII論文ID: 120005254091
• NII書誌ID: AA10681442
• ISSN: 02693879
• Web Site: http://hdl.handle.net/2297/34700
• 本文言語コード: en
• 資料種別: journal article
• データソース種別:
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  • CiNii Articles