CN111579514A - 一种硅酸锰模拟氧化酶催化氧化tmb的方法 - Google Patents
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- 238000000034 method Methods 0.000 title claims abstract description 16
- 102000004316 Oxidoreductases Human genes 0.000 title claims abstract description 15
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- 238000002835 absorbance Methods 0.000 claims description 3
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- 229910052760 oxygen Inorganic materials 0.000 abstract description 2
- 239000001301 oxygen Substances 0.000 abstract description 2
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 abstract 5
- 239000003054 catalyst Substances 0.000 abstract 1
- 238000010521 absorption reaction Methods 0.000 description 5
- 238000006555 catalytic reaction Methods 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 3
- 238000005034 decoration Methods 0.000 description 2
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- 238000002360 preparation method Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 1
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 1
- 239000004115 Sodium Silicate Substances 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000000378 calcium silicate Substances 0.000 description 1
- 229910052918 calcium silicate Inorganic materials 0.000 description 1
- OYACROKNLOSFPA-UHFFFAOYSA-N calcium;dioxido(oxo)silane Chemical compound [Ca+2].[O-][Si]([O-])=O OYACROKNLOSFPA-UHFFFAOYSA-N 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
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- 230000002068 genetic effect Effects 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- HCWCAKKEBCNQJP-UHFFFAOYSA-N magnesium orthosilicate Chemical compound [Mg+2].[Mg+2].[O-][Si]([O-])([O-])[O-] HCWCAKKEBCNQJP-UHFFFAOYSA-N 0.000 description 1
- 239000000391 magnesium silicate Substances 0.000 description 1
- 229910052919 magnesium silicate Inorganic materials 0.000 description 1
- 235000019792 magnesium silicate Nutrition 0.000 description 1
- 229940099607 manganese chloride Drugs 0.000 description 1
- 235000002867 manganese chloride Nutrition 0.000 description 1
- 239000011565 manganese chloride Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229910052914 metal silicate Inorganic materials 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 229910052911 sodium silicate Inorganic materials 0.000 description 1
- NTHWMYGWWRZVTN-UHFFFAOYSA-N sodium silicate Chemical compound [Na+].[Na+].[O-][Si]([O-])=O NTHWMYGWWRZVTN-UHFFFAOYSA-N 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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Abstract
本发明公开了一种利用硅酸锰纳米颗粒作为模拟氧化酶催化氧化TMB（3,3',5,5'‑四甲基联苯胺）的方法，包括如下步骤：室温下，于TMB溶液中加入硅酸锰纳米颗粒作为催化剂，在一定pH范围内直接催化氧气氧化TMB为蓝绿色物质，是一种可以替代天然酶催化氧化TMB的方法。本发明属于模拟酶催化氧化技术领域，该方法pH响应迅速、氧化效果显著，在生物催化氧化、医学免疫检测等领域具有重要的应用前景。
Description
技术领域
本发明提供一种利用硅酸锰纳米颗粒作为模拟氧化酶催化氧化TMB的方法，属于模拟酶催化氧化技术领域。
背景技术
酶作为一种天然的催化介质，在生物体内维持着遗传信息的传递和体内新陈代谢。生物体内的天然酶大多由蛋白质构成，具备专一性、催化效率高等优点，但其存在催化条件苛刻、极易失活变性、体外制备成本昂贵等固有缺点，阻碍了其进一步应用和发展。相比于天然酶，通过化学方法合成的、具有天然酶活性的、非蛋白质结构的模拟酶，有着催化条件温和、稳定性高、制备简单、成本较低等显著优点，可以大规模应用于生物催化、免疫检测、医学等领域。
金属硅酸盐材料（硅酸锰、硅酸钙、硅酸镁等）具备结构稳定、比表面积大、活性位点多等独特的物化性质，广泛应用于工业催化、能量转换与存储、纳米药物输运等领域。然而，现有技术中还未有关于将硅酸锰作为模拟氧化酶用于催化氧化TMB的报道。本发明提供了一种利用硅酸锰纳米颗粒作为模拟氧化酶催化氧化TMB的方法，该方法pH响应迅速、氧化效果显著，在生物催化氧化、医学免疫检测等领域具有重要的应用前景。
发明内容
本发明的目的是提供一种利用硅酸锰纳米颗粒作为模拟氧化酶催化氧化TMB的方法，可在一定pH范围内迅速催化溶液中的氧气氧化TMB。
为实现上述目的，本发明所采用的技术方案步骤如下：
一种利用硅酸锰纳米颗粒作为模拟氧化酶催化氧化TMB的方法，具体包括以下步骤：
（1）将一定量的TMB溶于无水乙醇中形成溶液Ⅰ。
上述方案（1）特征在于，溶液Ⅰ中TMB的浓度为32 mg ml-1。
（2）取0.5 ml溶液Ⅰ分散于7.5 ml磷酸盐缓冲溶液中，形成溶液Ⅱ。
上述方案（2）特征在于，溶液Ⅱ中磷酸盐缓冲溶液pH范围为5.4-7.4。
（3）将硅酸锰纳米颗粒加入溶液Ⅱ中，室温、避光下震荡使其分散均匀后孵育5分钟得溶液Ⅲ，随后将溶液Ⅲ放入紫外分光光度计中观察其吸光度。
上述方案（3）特征在于，硅酸锰纳米颗粒的浓度为0.2 mg ml-1。
相比于其他模拟氧化酶，本发明的特点在于：
（1）硅酸锰纳米颗粒作为模拟氧化酶催化氧化TMB的方法，在较宽pH范围内保持模拟氧化酶活性，催化TMB显色时间短，催化效率高。
（2）硅酸锰纳米颗粒模拟氧化酶在酸性条件下保持高催化活性，具有更高的化学稳定性。
附图说明
图1是实施例1所制备的硅酸锰纳米颗粒在pH范围5.4-7.4时催化氧化TMB的紫外吸收光谱。a曲线为在pH=5.4下测得的紫外吸收曲线；b曲线为在pH=6.0下测得的紫外吸收曲线；c曲线为在pH=6.4下测得的紫外吸收曲线；d曲线为在pH=7.0下测得的紫外吸收曲线;e曲线为在pH=7.4下测得的紫外吸收曲线。
具体实施方式
以下结合实施例对本发明作进一步说明。
实施例1
取1.42g硅酸钠和0.94g氯化锰溶于40mL去离子水中，室温下搅拌10分钟，过滤、烘干即得棕灰色硅酸锰纳米颗粒。
将本实施例所得硅酸锰纳米颗粒用于催化TMB显色，具体步骤包括：将16 mg TMB溶于0.5 ml无水乙醇中形成混合溶液I，取上述溶液I 0.5 ml分散于7.5 ml不同pH值的磷酸盐缓冲液中（pH分别为7.4、7.0、6.4、6.0、5.4）形成溶液Ⅱ，将1.6 mg硅酸锰纳米颗粒加入上述溶液Ⅱ中，在室温下避光、震荡使其分散均匀后孵育5分钟得溶液Ⅲ，利用紫外分光光度计（扫描652nm处），观察所得溶液体系Ⅲ的吸光度变化。
图1为本实施例所得硅酸锰纳米颗粒在不同pH条件下对TMB的催化结果图，图中可以看出，当磷酸盐缓冲液pH值为7.4时，本实施例所得纳米硅酸锰不可以催化TMB显色；随着磷酸盐缓冲液pH值降低，本实施例所得纳米硅酸锰对TMB的催化效果越来越好，表现出一种pH响应的催化效果。
以上所述仅是本发明的优选实施方式，本技术领域的普通技术人员应当了解，本发明不受实施例限制，还可以做出若干修改和润饰，这些修改和润饰也在本发明要求的保护范围内。
Claims (4)
1.一种利用硅酸锰纳米颗粒作为模拟氧化酶催化氧化TMB的方法，其特征在于，包括如下步骤：
（1）将一定量的TMB溶于无水乙醇中形成溶液Ⅰ；
（2）取0.5 ml溶液Ⅰ分散于7.5 ml磷酸盐缓冲溶液中，形成溶液Ⅱ；
（3）将上述纳米硅酸锰模拟氧化酶加入溶液Ⅱ中，室温、避光下震荡使其分散均匀后孵育5分钟得溶液Ⅲ，随后将溶液Ⅲ放入紫外分光光度计中观察其吸光度。
2.根据权利要求1所述的方法，其特征在于，所述的溶液Ⅰ中TMB的浓度为32 mg ml-1。
3.根据权利要求1所述的方法，其特征在于，溶液Ⅱ中磷酸盐缓冲溶液的pH范围为5.4-7.4。
4.根据权利要求1所述的方法，其特征在于，纳米硅酸锰模拟氧化酶得浓度为0.2 mgml-1。
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| CN113603106A (zh) * | 2021-09-01 | 2021-11-05 | 南京大学 | 基于硅酸盐溶解动力学制备硅酸锰纳米酶的方法及应用 |
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| CN113603106A (zh) * | 2021-09-01 | 2021-11-05 | 南京大学 | 基于硅酸盐溶解动力学制备硅酸锰纳米酶的方法及应用 |
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