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An analytical method for measuring α-amylase activity in starch-containing foods.
MedLine Citation:
PMID:  23074083     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
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 10 mU/mL, which is enough to reduce the viscosity of starch-containing foods. Copyright © 2012 John Wiley & Sons, Ltd.
Authors:
Kazuo Koyama; Takashi Hirao; Akira Toriba; Kazuichi Hayakawa
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-10-17
Journal Detail:
Title:  Biomedical chromatography : BMC     Volume:  -     ISSN:  1099-0801     ISO Abbreviation:  Biomed. Chromatogr.     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-10-17     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8610241     Medline TA:  Biomed Chromatogr     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2012 John Wiley & Sons, Ltd.
Affiliation:
Somatech Center, House Foods Corporation, 1-4 Takanodai, Yotsukaido-shi, Chiba, 284-0033, Japan; Graduate School of Natural Science and Technology, Institute of Medical Pharmaceutical and Health Science, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan. k-koyama@housefoods.co.jp.
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