JP5359169B2 - セルラーゼ測定試薬およびセルラーゼの測定方法 - Google Patents
セルラーゼ測定試薬およびセルラーゼの測定方法 Download PDFInfo
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- JP5359169B2 JP5359169B2 JP2008261575A JP2008261575A JP5359169B2 JP 5359169 B2 JP5359169 B2 JP 5359169B2 JP 2008261575 A JP2008261575 A JP 2008261575A JP 2008261575 A JP2008261575 A JP 2008261575A JP 5359169 B2 JP5359169 B2 JP 5359169B2
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- 108010059892 Cellulase Proteins 0.000 title claims abstract description 64
- 229940106157 cellulase Drugs 0.000 title claims abstract description 64
- 238000000034 method Methods 0.000 title claims abstract description 28
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- 229920002678 cellulose Polymers 0.000 claims abstract description 49
- 239000001913 cellulose Substances 0.000 claims abstract description 49
- 150000004696 coordination complex Chemical class 0.000 claims abstract description 26
- 229910052751 metal Inorganic materials 0.000 claims abstract description 25
- 239000002184 metal Substances 0.000 claims abstract description 25
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- 239000008103 glucose Substances 0.000 claims abstract description 9
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims abstract description 7
- 238000005259 measurement Methods 0.000 claims description 16
- IEQIEDJGQAUEQZ-UHFFFAOYSA-N phthalocyanine Chemical compound N1C(N=C2C3=CC=CC=C3C(N=C3C4=CC=CC=C4C(=N4)N3)=N2)=C(C=CC=C2)C2=C1N=C1C2=CC=CC=C2C4=N1 IEQIEDJGQAUEQZ-UHFFFAOYSA-N 0.000 claims description 2
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 229930182470 glycoside Natural products 0.000 abstract description 2
- 239000000413 hydrolysate Substances 0.000 abstract 1
- 230000003301 hydrolyzing effect Effects 0.000 abstract 1
- 238000010521 absorption reaction Methods 0.000 description 9
- 229910052802 copper Inorganic materials 0.000 description 8
- 239000010949 copper Substances 0.000 description 8
- 239000000523 sample Substances 0.000 description 8
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 7
- 229920000742 Cotton Polymers 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 6
- RBTKNAXYKSUFRK-UHFFFAOYSA-N heliogen blue Chemical compound [Cu].[N-]1C2=C(C=CC=C3)C3=C1N=C([N-]1)C3=CC=CC=C3C1=NC([N-]1)=C(C=CC=C3)C3=C1N=C([N-]1)C3=CC=CC=C3C1=N2 RBTKNAXYKSUFRK-UHFFFAOYSA-N 0.000 description 6
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- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
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- XCJYREBRNVKWGJ-UHFFFAOYSA-N copper(II) phthalocyanine Chemical compound [Cu+2].C12=CC=CC=C2C(N=C2[N-]C(C3=CC=CC=C32)=N2)=NC1=NC([C]1C=CC=CC1=1)=NC=1N=C1[C]3C=CC=CC3=C2[N-]1 XCJYREBRNVKWGJ-UHFFFAOYSA-N 0.000 description 3
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- AZQWKYJCGOJGHM-UHFFFAOYSA-N 1,4-benzoquinone Chemical compound O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
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- 239000000126 substance Substances 0.000 description 2
- VEUMANXWQDHAJV-UHFFFAOYSA-N 2-[2-[(2-hydroxyphenyl)methylideneamino]ethyliminomethyl]phenol Chemical compound OC1=CC=CC=C1C=NCCN=CC1=CC=CC=C1O VEUMANXWQDHAJV-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 235000012766 Cannabis sativa ssp. sativa var. sativa Nutrition 0.000 description 1
- 235000012765 Cannabis sativa ssp. sativa var. spontanea Nutrition 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 244000146553 Ceiba pentandra Species 0.000 description 1
- 235000003301 Ceiba pentandra Nutrition 0.000 description 1
- 229920000298 Cellophane Polymers 0.000 description 1
- 108010084185 Cellulases Proteins 0.000 description 1
- 102000005575 Cellulases Human genes 0.000 description 1
- 241000218631 Coniferophyta Species 0.000 description 1
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-CUHNMECISA-N 0.000 description 1
- 241001265525 Edgeworthia chrysantha Species 0.000 description 1
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- 229920000663 Hydroxyethyl cellulose Polymers 0.000 description 1
- 239000004354 Hydroxyethyl cellulose Substances 0.000 description 1
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- 235000008708 Morus alba Nutrition 0.000 description 1
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- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 229920002522 Wood fibre Polymers 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- ZIALXKMBHWELGF-UHFFFAOYSA-N [Na].[Cu] Chemical compound [Na].[Cu] ZIALXKMBHWELGF-UHFFFAOYSA-N 0.000 description 1
- CBHOOMGKXCMKIR-UHFFFAOYSA-N azane;methanol Chemical compound N.OC CBHOOMGKXCMKIR-UHFFFAOYSA-N 0.000 description 1
- 229920001222 biopolymer Polymers 0.000 description 1
- 235000009120 camo Nutrition 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 235000005607 chanvre indien Nutrition 0.000 description 1
- 239000013626 chemical specie Substances 0.000 description 1
- 229910052804 chromium Inorganic materials 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
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- 230000000694 effects Effects 0.000 description 1
- 238000000921 elemental analysis Methods 0.000 description 1
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- 238000006911 enzymatic reaction Methods 0.000 description 1
- -1 etc.) Polymers 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000011121 hardwood Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 239000011487 hemp Substances 0.000 description 1
- 235000019447 hydroxyethyl cellulose Nutrition 0.000 description 1
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 description 1
- 229910001410 inorganic ion Inorganic materials 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 229910052748 manganese Inorganic materials 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000004032 porphyrins Chemical class 0.000 description 1
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- WXQMFIJLJLLQIS-UHFFFAOYSA-N reactive blue 21 Chemical compound [Cu+2].C1=CC(S(=O)(=O)CCO)=CC=C1NS(=O)(=O)C1=CC=C2C([N-]3)=NC(C=4C5=CC=C(C=4)S(O)(=O)=O)=NC5=NC(C=4C5=CC=C(C=4)S(O)(=O)=O)=NC5=NC([N-]4)=C(C=C(C=C5)S(O)(=O)=O)C5=C4N=C3C2=C1 WXQMFIJLJLLQIS-UHFFFAOYSA-N 0.000 description 1
- 239000004627 regenerated cellulose Substances 0.000 description 1
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- 229910052720 vanadium Inorganic materials 0.000 description 1
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- 229910052727 yttrium Inorganic materials 0.000 description 1
- VWQVUPCCIRVNHF-UHFFFAOYSA-N yttrium atom Chemical compound [Y] VWQVUPCCIRVNHF-UHFFFAOYSA-N 0.000 description 1
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Description
(1)セルラーゼ測定試薬(金属錯体結合型セルロース)の調製:
炭酸ナトリウム:2.0g、硫酸ナトリウム:5.0g、C.I.Reactive Blue:21:0.6gを溶かした水溶液100mLに脱脂綿5.0gを添加し、攪拌しながら30℃で35分反応させた。その後、70℃にて60分間で反応させ、反応の後、ジメチルスルホキシド、メタノール-アンモニア(50:1v/v)混合溶液で洗浄し、乾燥させて金属錯体結合型セルロースを得た。この金属錯体結合型セルロースにおいて、金属錯体の割合は、金属錯体結合型セルロース1.0g当たり30μmolであり、セルロースのグルコース約600個当たり1個であった。
濃度の異なる複数のセルラーゼ水溶液を調製して試料とした。20mLサンプル管に各10mLを加え、恒温槽にて45℃の一定温度に保持した。その後、上記のセルラーゼ測定試薬0.02gを加え、1時間振とうした。振とうの後、上澄み溶液を分取し、フレームレス原子吸光光度計(日立ハイテクノロジー社製「 Z−2000型」)にて、試料溶液を20μL注入し、銅の分析を行った。
実施例1における銅の分析において、フレームレス原子吸光光度計に代えてICP発光分光分析装置(パーキンエルマー社製 「Optima5300DV型」)を使用した他は、実施例1と同様に行った。その結果、銅の分析値は、フレームレス原子吸光光度計を使用した実施例1と同様の値であった。なお、ICP発光分光分析装置における測定条件は次の通りである。すなわち、試料溶液と80ppbイットリウム内標準溶液(5%硝酸溶液)をオンライン内標準添加法にて流路内にて混合した。その際の吸引流速は0.25mL/minとした。
Claims (3)
- セルロースの構成グルコースの水酸基に反応基を介して金属錯体を結合した金属錯体結合型セルロースから成ることを特徴とするセルラーゼ測定試薬。
- 金属錯体が金属フタロシアニンである請求項1に記載のセルラーゼ測定試薬。
- 請求項1又は2に記載のセルラーゼ測定試薬にセルラーゼを作用させてセルロースのグリコシド結合を加水分解し、生成する加水分解物に固定された金属錯体中の金属の濃度に基づき、セルラーゼの濃度を測定することを特徴とするセルラーゼの測定方法。
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JP2010088349A JP2010088349A (ja) | 2010-04-22 |
JP5359169B2 true JP5359169B2 (ja) | 2013-12-04 |
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MA25183A1 (fr) * | 1996-05-17 | 2001-07-02 | Arthur Jacques Kami Christiaan | Compositions detergentes |
JP3827465B2 (ja) * | 1999-02-26 | 2006-09-27 | 大日精化工業株式会社 | 改良された化学発光試薬 |
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