JPS6123995B2 - - Google Patents
Info
- Publication number
- JPS6123995B2 JPS6123995B2 JP55145881A JP14588180A JPS6123995B2 JP S6123995 B2 JPS6123995 B2 JP S6123995B2 JP 55145881 A JP55145881 A JP 55145881A JP 14588180 A JP14588180 A JP 14588180A JP S6123995 B2 JPS6123995 B2 JP S6123995B2
- Authority
- JP
- Japan
- Prior art keywords
- flavin
- gpo
- albumin
- solution
- containing substance
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 claims description 19
- 239000000126 substance Substances 0.000 claims description 19
- 239000000203 mixture Substances 0.000 claims description 18
- 108010088751 Albumins Proteins 0.000 claims description 12
- 102000009027 Albumins Human genes 0.000 claims description 12
- 150000001413 amino acids Chemical class 0.000 claims description 11
- 235000000346 sugar Nutrition 0.000 claims description 7
- 150000001875 compounds Chemical class 0.000 claims description 5
- 150000008163 sugars Chemical class 0.000 claims description 5
- 108010041921 Glycerolphosphate Dehydrogenase Proteins 0.000 claims description 4
- 102000000587 Glycerolphosphate Dehydrogenase Human genes 0.000 claims description 4
- 229940024606 amino acid Drugs 0.000 description 10
- 235000001014 amino acid Nutrition 0.000 description 10
- 239000000243 solution Substances 0.000 description 10
- 239000007864 aqueous solution Substances 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 239000000654 additive Substances 0.000 description 5
- 150000001720 carbohydrates Chemical class 0.000 description 4
- 238000004108 freeze drying Methods 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 239000006172 buffering agent Substances 0.000 description 3
- 235000019162 flavin adenine dinucleotide Nutrition 0.000 description 3
- 239000011714 flavin adenine dinucleotide Substances 0.000 description 3
- VWWQXMAJTJZDQX-UYBVJOGSSA-N flavin adenine dinucleotide Chemical compound C1=NC2=C(N)N=CN=C2N1[C@@H]([C@H](O)[C@@H]1O)O[C@@H]1CO[P@](O)(=O)O[P@@](O)(=O)OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C2=NC(=O)NC(=O)C2=NC2=C1C=C(C)C(C)=C2 VWWQXMAJTJZDQX-UYBVJOGSSA-N 0.000 description 3
- 229940093632 flavin-adenine dinucleotide Drugs 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 239000003381 stabilizer Substances 0.000 description 3
- RLFWWDJHLFCNIJ-UHFFFAOYSA-N 4-aminoantipyrine Chemical compound CN1C(C)=C(N)C(=O)N1C1=CC=CC=C1 RLFWWDJHLFCNIJ-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- GNGACRATGGDKBX-UHFFFAOYSA-N dihydroxyacetone phosphate Chemical compound OCC(=O)COP(O)(O)=O GNGACRATGGDKBX-UHFFFAOYSA-N 0.000 description 2
- 150000002016 disaccharides Chemical class 0.000 description 2
- 229940013640 flavin mononucleotide Drugs 0.000 description 2
- 239000011768 flavin mononucleotide Substances 0.000 description 2
- FVTCRASFADXXNN-SCRDCRAPSA-N flavin mononucleotide Chemical compound OP(=O)(O)OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O FVTCRASFADXXNN-SCRDCRAPSA-N 0.000 description 2
- FVTCRASFADXXNN-UHFFFAOYSA-N flavin mononucleotide Natural products OP(=O)(O)OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O FVTCRASFADXXNN-UHFFFAOYSA-N 0.000 description 2
- 108010054790 glycerol-3-phosphate oxidase Proteins 0.000 description 2
- -1 lactose and sucrose Chemical compound 0.000 description 2
- 150000002772 monosaccharides Chemical class 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 235000019231 riboflavin-5'-phosphate Nutrition 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose 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)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 108091006905 Human Serum Albumin Proteins 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 125000004072 flavinyl group Chemical group 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- AWUCVROLDVIAJX-VKHMYHEASA-N sn-glycerol 1-phosphate Chemical compound OC[C@H](O)COP(O)(O)=O AWUCVROLDVIAJX-VKHMYHEASA-N 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- HFQQZARZPUDIFP-UHFFFAOYSA-M sodium;2-dodecylbenzenesulfonate Chemical compound [Na+].CCCCCCCCCCCCC1=CC=CC=C1S([O-])(=O)=O HFQQZARZPUDIFP-UHFFFAOYSA-M 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
Description
本発明は安定化されたグリセロリン酸オキシダ
ーゼ組成物に関するものである。
近年、Ｌ−α−グリセロリン酸を酸化してジヒ
ドロキシアセトンリン酸と過酸化水素を生成する
Ｌ−α−グリセロリン酸オキシダーゼ（以下
GPOと略称する）は診断用試薬として使用さ
れ、その重要性がますます増加してきた。ところ
がGPOは不安定であり、特にGPOを水溶液にし
た場合、該水溶液中のGPO濃度が１mg／ml以下
の低濃度では極端に不安定である。さらにGPO
を含有する組成物は凍結乾燥に極めて敏感で弱
く、GPO活性は凍結乾燥の間、あるいは凍結乾
燥後、相当低下する。すたがつて、低濃度の
GPO水溶液では長期間に亘る品質の維持が困難
である。このような理由からGPOを用いる利点
が失われる場合が多い。
本発明者等はこのような欠点を有しない安定な
GPO含有組成物を得るべく、種々鋭意検討した
結果、安定化剤として、フラビン含有物質又はフ
ラビン含有物と糖類、アルブミンまたはアミノ酸
を使用すると所期の目的を達成することを見出
し、本発明に到達した。すなわち本発明は(i)フラ
ビン含有物質又は(ii)糖類、アルブミンおよびアミ
ノ酸からなる群から選択された１種または２種以
上の化合物とフラビン含有物質および(iii)GPOを
含有する水性組成物またはその凍結乾燥物からな
る安定化されたグリセロリン酸オキシダーゼ組成
物である。
本発明では、上記安定剤を使用することによ
り、低濃度の水溶液においても、あるいは凍結乾
燥によつてもGPOの活性低下が見られない。さ
らに、安定化効果は長期間に亘つて、持続するこ
とができる。
本発明において使用するフラビン含有物質とし
ては、フラビン構造を含有する化合物であればよ
く、例えばビタミンB2、フラビンアデニンジヌ
クレオチド（FAD）、フラビンモノヌクレオチド
（FMN）等の物質およびそれらの塩がある。特に
FADが好ましい。
糖類としてはブドウ糖、ソルビトール、マンニ
トールなどの単糖類、乳糖、シヨ糖などの二糖
類、可溶性デンプンなどの多糖類が挙げられ、好
ましくは単糖類または二糖類である。
アルブミンとしてはヒト血清アルブミン、ウマ
血清アルブミン、ウシ血清アルブミン、羊血清ア
ルブミンなどの血清アルブミンが挙げられ、好ま
しくはウシ血清アルブミンである。
アミノ酸としてはグリシン、アラニン、リジ
ン、アルギニン、トリプトフアン、ロイシン、イ
ソロイシンなどがある。
本発明の水性組成物は緩衝液にGPOと(i)フラ
ビン含有物質又は(ii)糖類、アルブミンおよびアミ
ノ酸からなる群から選択された１種又は２種以上
の化合物とフラビン含有物質を含有させたもので
ある。緩衝剤としては通常のものが使用され、該
水性組成物のPHを通常５〜10とするものが好まし
い。
本発明の水性組成物には必要により、他の成分
を添加してもよい。
本発明の水性組成物において、フラビン含有物
質の含有量は１×10-6〜１×10-2Mである。糖類
の含有量は通常、該水性組成物１ml当り100×
10-6〜50×10-3ｇである。アルブミンの含有量は
通常、該水性組成物１ml当り、１×10-6〜100×
10-3ｇである。アミノ酸の含有量は通常、該水性
組成物１ml当り、100×10-6〜100×10-31ｇであ
る。
安定化剤としては上記化合物のうち、２種以上
を用いるとが好ましく、特にフラビン含有物質と
アルブミン、糖類又はアミノ酸との併用が好まし
い。
フラビン含有物質、糖類、アルブミン、アミノ
酸をそれぞれ併用する場合、含有量は各々の含有
量の和となる量でよい。
水性組成物を調製する場合、緩衝液への各物質
の添加順序は特に制限されない。
本発明の水性組成物は通常の方法に従い、凍結
乾燥して凍結乾燥物を得る。
本発明では溶液状態（水性組成物）である場合
にも、凍結乾燥状態（凍結乾燥物）である場合に
も、GPOの安定性は著しく高く、長期間に亘つ
て安定性が維持される。
次に本発明を実施例により説明する。
なおGPOの酵素活性の測定は次の方法に従つ
た。0.2Mトリス塩酸緩衝液（PH8.0）0.2ml、パー
オキシターゼ（340単位／ml）0.1ml、４−アミノ
アンチピリン（0.3％水溶液）0.1ml、フエノール
（0.2水溶液）0.1ml、DL−グリセロール−３−リ
ン酸ナトリウム（0.5M PH8.0）0.2ml、トリトン
Ｘ−100（0.5％水溶液）0.1mlおよび水0.2mlを混
合し、37℃で５分間加温した後、20μの酵素溶
解液を加え、37℃で10分間加温した。次いで１％
ラウリルベンゼンスルホン酸ソーダ水溶液0.5ml
を加えて反応を停止さ、さらに水1.5mlを加えて
500nmの吸光度を測定した。
比活性（Ｕ／mg）＝△Ａ／１０／１３．３×１／２×
３．０２／０．０２×１／Ｘ
Ｘ：酵素溶解液中の酵素濃度
△Ａ：As−Ac
As：酵素溶解液を用いたときの吸光度
Ac：酵素溶解液に代えて水を用いたときの吸
光度
実施例 １
GPOを0.05Mリン酸緩衝液（PH7.0）に溶解し
た。溶液中のGPO濃度は0.03mg／mlであつた。下
記表の添加物を該溶液に加え、４℃にて４週間、
さらに25℃にて１週間保存し、安定性を検討し
た。その結果を第１表および第２表に示す。溶液
調製直後の酵素活性を100％とした。
(1)４℃、４週間
The present invention relates to stabilized glycerophosphate oxidase compositions. In recent years, L-α-glycerophosphate oxidase (hereinafter referred to as L-α-glycerophosphate oxidase), which oxidizes L-α-glycerophosphate to produce dihydroxyacetone phosphate and hydrogen peroxide, has been developed.
GPO) is used as a diagnostic reagent and has become increasingly important. However, GPO is unstable, and especially when GPO is made into an aqueous solution, it is extremely unstable if the GPO concentration in the aqueous solution is as low as 1 mg/ml or less. More GPOs
Compositions containing GPO are extremely sensitive to lyophilization and GPO activity decreases considerably during or after lyophilization. stiff, low concentration
It is difficult to maintain the quality of GPO aqueous solutions over a long period of time. For these reasons, the benefits of using GPOs are often lost. The inventors have proposed a stable method that does not have these drawbacks.
As a result of various intensive studies in order to obtain a GPO-containing composition, it was discovered that the intended purpose could be achieved by using a flavin-containing substance or a flavin-containing substance and a saccharide, albumin, or an amino acid as a stabilizer, and the present invention was achieved. did. That is, the present invention provides an aqueous composition containing (i) a flavin-containing substance, or (ii) one or more compounds selected from the group consisting of sugars, albumin, and amino acids, a flavin-containing substance, and (iii) GPO; This is a stabilized glycerophosphate oxidase composition consisting of a lyophilized product thereof. In the present invention, by using the above-mentioned stabilizer, no decrease in GPO activity is observed even in a low concentration aqueous solution or by freeze-drying. Furthermore, the stabilizing effect can last for a long time. The flavin-containing substance used in the present invention may be any compound containing a flavin structure, such as vitamin B 2 , flavin adenine dinucleotide (FAD), flavin mononucleotide (FMN), and salts thereof. . especially
FAD is preferred. Examples of saccharides include monosaccharides such as glucose, sorbitol, and mannitol, disaccharides such as lactose and sucrose, and polysaccharides such as soluble starch, and preferably monosaccharides or disaccharides. Examples of the albumin include serum albumins such as human serum albumin, horse serum albumin, bovine serum albumin, and sheep serum albumin, with bovine serum albumin being preferred. Amino acids include glycine, alanine, lysine, arginine, tryptophan, leucine, and isoleucine. The aqueous composition of the present invention contains GPO and (i) a flavin-containing substance, or (ii) one or more compounds selected from the group consisting of sugars, albumin, and amino acids and a flavin-containing substance in a buffer solution. It is something. As the buffering agent, a conventional buffering agent is used, and it is preferable that the buffering agent adjusts the pH of the aqueous composition to usually 5 to 10. If necessary, other components may be added to the aqueous composition of the present invention. In the aqueous composition of the present invention, the content of the flavin-containing substance is 1×10 −6 to 1×10 −2 M. The sugar content is usually 100x per ml of the aqueous composition.
10 −6 to 50×10 −3 g. The content of albumin is usually 1×10 -6 to 100× per ml of the aqueous composition.
10 -3 g. The content of amino acids is usually 100×10 −6 to 100×10 −3 1 g per ml of the aqueous composition. As the stabilizer, it is preferable to use two or more of the above compounds, and it is particularly preferable to use a flavin-containing substance in combination with albumin, saccharide, or amino acid. When flavin-containing substances, saccharides, albumin, and amino acids are used in combination, the content may be the sum of the respective contents. When preparing an aqueous composition, the order of adding each substance to the buffer is not particularly limited. The aqueous composition of the present invention is freeze-dried to obtain a freeze-dried product according to a conventional method. In the present invention, the stability of GPO is extremely high whether it is in a solution state (aqueous composition) or in a freeze-dried state (freeze-dried product), and the stability is maintained over a long period of time. Next, the present invention will be explained by examples. The enzymatic activity of GPO was measured according to the following method. 0.2M Tris-HCl buffer (PH8.0) 0.2ml, peroxidase (340 units/ml) 0.1ml, 4-aminoantipyrine (0.3% aqueous solution) 0.1ml, phenol (0.2aqueous solution) 0.1ml, DL-glycerol-3 - Mix 0.2 ml of sodium phosphate (0.5M PH8.0), 0.1 ml of Triton , and warmed at 37°C for 10 minutes. Then 1%
Sodium laurylbenzenesulfonate aqueous solution 0.5ml
Add 1.5 ml of water to stop the reaction, then add 1.5 ml of water.
Absorbance at 500 nm was measured. Specific activity (U/mg)=△A/10/13.3×1/2×
3.02/0.02×1/X Absorbance Example 1 GPO was dissolved in 0.05M phosphate buffer (PH7.0). The GPO concentration in the solution was 0.03 mg/ml. The additives listed in the table below were added to the solution and incubated at 4°C for 4 weeks.
It was further stored at 25°C for one week to examine stability. The results are shown in Tables 1 and 2. Enzyme activity immediately after solution preparation was defined as 100%. (1) 4℃, 4 weeks
【表】【table】
【表】 (2)25℃、１週間【table】 (2) 25℃, 1 week
【表】【table】
【表】
(1)，(2)のいずれの保存においても、フラビン含
有物質又はフラビン含有物質と糖類、アルブミン
又はアミノ酸を添加した場合、GPOの安定性が
著しく向上した。
実施例 ２
GPOを0.05Mリン酸緩衝液（PH7.0）に溶解し
た。溶液中のGPO濃度は0.5mg／mlであつた。下
記表の添加物を該溶液に加え、凍結乾燥し、安定
性を検討した。その結果を第３表に示す。溶液調
製直後の酵素活性を100％とした。[Table] In both storage methods (1) and (2), the stability of GPO was significantly improved when flavin-containing substances or flavin-containing substances and sugars, albumin, or amino acids were added. Example 2 GPO was dissolved in 0.05M phosphate buffer (PH7.0). The GPO concentration in the solution was 0.5 mg/ml. Additives listed in the table below were added to the solution, freeze-dried, and examined for stability. The results are shown in Table 3. Enzyme activity immediately after solution preparation was defined as 100%.
【表】【table】
【表】
フラビン含有物質又はフラビン含有物質と糖
類、アルブミン又はアミノ酸を添加して凍結乾燥
した場合、無添加に比較して著しく失活が少な
い。
実施例 ３
下記表の添加物を加えたGPO溶液を凍結乾燥
した凍結乾燥品の、40℃、１週間での安定性を検
討した。その結果を第４表に示す。溶液調製直後
の酵素活性を100％とした。[Table] When a flavin-containing substance or a flavin-containing substance and a sugar, albumin, or amino acid are added and freeze-dried, there is significantly less deactivation than when no additive is added. Example 3 The stability of a freeze-dried product obtained by freeze-drying a GPO solution containing the additives shown in the table below at 40°C for one week was investigated. The results are shown in Table 4. Enzyme activity immediately after solution preparation was defined as 100%.
【表】【table】
【表】
フラビン含有物質又はフラビン含有物質と糖
類、アルブミン又はアミノ酸を添加した凍結乾燥
品は無添加に比べ著しく安定化された。[Table] Freeze-dried products to which flavin-containing substances or flavin-containing substances and sugars, albumin, or amino acids were added were significantly more stable than those without any additives.
Claims (1)
よびアミノ酸からなる群から選択された１種又は
２種以上の化合物とフラビン含有物質および(iii)グ
リセロリン酸オキシダーゼを含有する水性組成物
またはその凍結乾燥物からなる安定化されたグリ
セロリン酸オキシダーゼ組成物。1. An aqueous composition containing a flavin-containing substance or (ii) one or more compounds selected from the group consisting of sugars, albumin, and amino acids, a flavin-containing substance, and (iii) glycerophosphate oxidase, or a lyophilized product thereof. A stabilized glycerophosphate oxidase composition comprising:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14588180A JPS5768788A (en) | 1980-10-17 | 1980-10-17 | Stablized glycerophosphate oxidase composition |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14588180A JPS5768788A (en) | 1980-10-17 | 1980-10-17 | Stablized glycerophosphate oxidase composition |
Related Child Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP25817184A Division JPS60156386A (en) | 1984-12-05 | 1984-12-05 | Stabilized glycerophosphate oxidase composition |
| JP9607287A Division JPS63263082A (en) | 1987-04-17 | 1987-04-17 | Stabilized glycerophosphoric acid oxidase |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5768788A JPS5768788A (en) | 1982-04-27 |
| JPS6123995B2 true JPS6123995B2 (en) | 1986-06-09 |
Family
ID=15395202
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP14588180A Granted JPS5768788A (en) | 1980-10-17 | 1980-10-17 | Stablized glycerophosphate oxidase composition |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5768788A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010022328A (en) * | 2008-07-23 | 2010-02-04 | Aisin Seiki Co Ltd | Method for stabilizing coenzyme-binding enzyme, and composition, enzyme sensor and fuel cell prepared by using the stabilization method |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4684615A (en) * | 1983-06-06 | 1987-08-04 | Ciba Corning Diagnostics Corp. | Stabilized isoenzyme control products |
| US4810633A (en) * | 1984-06-04 | 1989-03-07 | Miles Inc. | Enzymatic ethanol test |
| JPS63263082A (en) * | 1987-04-17 | 1988-10-31 | Toyobo Co Ltd | Stabilized glycerophosphoric acid oxidase |
| US5185247A (en) * | 1989-03-10 | 1993-02-09 | Miles Inc. | Stabilization of oxidase enzyme-based test strips |
| US5116729A (en) * | 1989-03-10 | 1992-05-26 | Miles Inc. | Stabilization of oxidase enzyme-based test strips |
| US5192657A (en) * | 1990-12-18 | 1993-03-09 | Ortho Diagnostic Systems, Inc. | Stabilized proteolytic solution and reagent kit |
| JPH0539376U (en) * | 1991-11-08 | 1993-05-28 | 株式会社ソーゴ | Seasoning stand |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS53103821A (en) * | 1977-02-21 | 1978-09-09 | Shingu Shoko Kk | Drawinggout device for cord in bush cleaneer * especially grass mower |
| JPS6123995A (en) * | 1984-07-12 | 1986-02-01 | 株式会社日立製作所 | Water-supply-pressure cooperation controller for nuclear reactor |
-
1980
- 1980-10-17 JP JP14588180A patent/JPS5768788A/en active Granted
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS53103821A (en) * | 1977-02-21 | 1978-09-09 | Shingu Shoko Kk | Drawinggout device for cord in bush cleaneer * especially grass mower |
| JPS6123995A (en) * | 1984-07-12 | 1986-02-01 | 株式会社日立製作所 | Water-supply-pressure cooperation controller for nuclear reactor |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010022328A (en) * | 2008-07-23 | 2010-02-04 | Aisin Seiki Co Ltd | Method for stabilizing coenzyme-binding enzyme, and composition, enzyme sensor and fuel cell prepared by using the stabilization method |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5768788A (en) | 1982-04-27 |
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