JP5532256B2 - 糖質定量方法および糖質定量キット - Google Patents
糖質定量方法および糖質定量キット Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
- C12Q1/40—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving amylase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
- C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/924—Hydrolases (3) acting on glycosyl compounds (3.2)
- G01N2333/926—Hydrolases (3) acting on glycosyl compounds (3.2) acting on alpha -1, 4-glucosidic bonds, e.g. hyaluronidase, invertase, amylase
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/924—Hydrolases (3) acting on glycosyl compounds (3.2)
- G01N2333/926—Hydrolases (3) acting on glycosyl compounds (3.2) acting on alpha -1, 4-glucosidic bonds, e.g. hyaluronidase, invertase, amylase
- G01N2333/934—Glucoamylase
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Description
具体的には、本発明の第一態様は消化酵素を用いた糖質定量方法であって、前記消化酵素として、動物由来の低分子糖質消化酵素が用いられ、前記低分子糖質消化酵素は、破砕した動物小腸を懸濁した小腸粘膜刷子縁膜の懸濁液からなることを特徴としている。
さらに詳細には、消化酵素を有し、イソマルトオリゴ糖、フラクトオリゴ糖、ラクトース、スクロースについて、小腸において消化される消化性糖質と、小腸において消化されない難消化性糖質とに区別して定量する糖質定量キットであって、前記消化酵素として、動物由来の低分子糖質消化酵素を備え、前記低分子糖質消化酵素は、切除して得られた動物小腸の上部を破砕し、遠心分離して得られた沈殿物を懸濁した小腸粘膜刷子縁の懸濁液からなることを特徴としている。
具体的には、本発明は、前記第一態様にかかる糖質定量方法であって、耐熱性α―アミラーゼを用いた第一反応工程と、プロテアーゼおよびアミログルコシダーゼを用いた第二反応工程と、動物由来の前記低分子糖質消化酵素を用いた第三反応工程とを備えたことを特徴としている。
具体的には、本発明は、消化酵素を有する糖質定量キットであって、前記消化酵素として、動物由来の低分子糖質消化酵素を備え、前記低分子糖質消化酵素は、破砕した動物小腸を懸濁した小腸粘膜刷子縁膜の懸濁液からなることを特徴としている。
図1は、本発明の第一実施例にかかる糖質定量方法の操作手順図を示したものである。より具体的には、図1は、本実施例にかかる酵素−HPLC法による糖質定量の操作手順図を示したものである。
カラム Shodex SUGAR KS−802
(8.0φ×300mm,昭和電工(株)製)
カラム温度 70℃
移動相 H2O
流速 0.5mL/min
試料流入量 10μL
検出器 示差屈折計 RID−10A((株)島津製作所製)
未消化物の回収率(%)=(((A/B)×希釈倍率)/試料重量(g))×100
ここで、A:酵素処理後の糖質のピークエリア
B:標準物質溶液のピークエリア
なお、本発明は、上記実施形態および実施例(以下「実施形態等」という。)に限定されるものではなく、必要に応じて種々の変更を加えて実施することも可能であり、それらはいずれも本発明の技術的範囲に含まれる。
Claims (1)
- 消化酵素を有し、イソマルトオリゴ糖、フラクトオリゴ糖、ラクトース、スクロースについて、小腸において消化される消化性糖質と、小腸において消化されない難消化性糖質とに区別して定量する糖質定量キットであって、
前記消化酵素として、動物由来の低分子糖質消化酵素を備え、
前記低分子糖質消化酵素は、切除して得られた動物小腸の上部を破砕し、遠心分離して得られた沈殿物を懸濁した小腸粘膜刷子縁の懸濁液からなる
ことを特徴とする糖質定量キット。
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/JP2008/058440 WO2009136432A1 (ja) | 2008-05-06 | 2008-05-06 | 糖質定量方法および糖質定量キット |
Publications (2)
Publication Number | Publication Date |
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JPWO2009136432A1 JPWO2009136432A1 (ja) | 2011-09-01 |
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JPH0873482A (ja) * | 1994-06-27 | 1996-03-19 | Hayashibara Biochem Lab Inc | 還元性を低減させた糖質とその製造方法並びに用途 |
JPH10150934A (ja) * | 1996-11-20 | 1998-06-09 | Matsutani Chem Ind Ltd | 還元難消化性水飴およびこれを用いた食品 |
WO2006035725A1 (ja) * | 2004-09-27 | 2006-04-06 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | イソサイクロマルトオリゴ糖及びイソサイクロマルトオリゴ糖生成酵素とそれらの製造方法並びに用途 |
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US3249514A (en) * | 1963-11-18 | 1966-05-03 | Bode Harold Eli | Production and use of amyloglucosidase |
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JP3183500B2 (ja) | 1989-04-10 | 2001-07-09 | 日本化薬株式会社 | 糖アルコールの定量方法 |
IL114030A (en) | 1994-06-27 | 2000-08-13 | Hayashibara Biochem Lab | Saccharide composition with a lesser reducibility its preparation and compositions containing it |
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JPH0873482A (ja) * | 1994-06-27 | 1996-03-19 | Hayashibara Biochem Lab Inc | 還元性を低減させた糖質とその製造方法並びに用途 |
JPH10150934A (ja) * | 1996-11-20 | 1998-06-09 | Matsutani Chem Ind Ltd | 還元難消化性水飴およびこれを用いた食品 |
WO2006035725A1 (ja) * | 2004-09-27 | 2006-04-06 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | イソサイクロマルトオリゴ糖及びイソサイクロマルトオリゴ糖生成酵素とそれらの製造方法並びに用途 |
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JPN6013004510; '日本食品科学工学会誌' 2004年、第51巻、第1号、第28頁-第33頁 * |
JPN6013004513; 'Analytical and Bioanalytical Chemistry' 2007, Vol.389, No.1, pp.291-308 * |
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US20110059474A1 (en) | 2011-03-10 |
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