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一种以酶为中心的方法,用于构建具有长和可编程线性范围的电压计 l-酸盐生物传感器

Christopher J Matthews¹, Wayne M Patrick¹

¹Centre for Biodiscovery, School of Biological Sciences, Victoria University of Wellington, Wellington, New Zealand.

Protein science : a publication of the Protein Society

|July 29, 2023

概括

研究人员开发了一种用于生物传感器的新酶,以准确测量未稀释的果汁和葡萄酒中的l-酸盐. 这种酶提供了一个可调节的范围,改进了现有的酸盐检测方法用于食品和饮料分析.

科学领域:

生物化学和生物传感器技术
酶工程和定向进化的指导进化.
食品和饮料的分析.

背景情况:

L-酸盐对于食品和饮料,特别是葡萄酒的风味和酸度至关重要.
目前对l-酸盐的酶生物传感器具有有限的线性范围,需要样品稀释.
需要更加多功能和灵敏的l-酸盐检测方法.

研究的目的:

发现和设计新的酸盐氧化酶,以提高生物传感器性能.
开发一种l-酸盐生物传感器,具有可调节的线性范围和高灵敏度.
在现实世界的食品和饮料样本中验证生物传感器的功能.

主要方法:

数据库挖掘,基因合成和重组表达以识别和产生酶.
光谱测量测试以描述酶活性和适用于生物传感器的适用性.
使用新型酶开发和优化一个安培度生物传感器原型.

主要成果:

一个定制的生物催化剂,Ascaris suum malic酶与突变R181Q [AsME(R181Q) ],被工程.
在最初的配置中,ASME (R181Q) 生物传感器显示了超宽线性范围 (50-200 mM).
对于Mn2+的敏感性增加了五倍,对于酸盐的敏感性增加了六倍,产生了1-10mM的线性范围.
使用酸盐氧化酶减轻了酸盐的干扰,从而可以在未稀释的葡萄酒中进行准确的测量.

结论:

一种以酶为中心的方法成功产生了一种具有可适应灵敏度和线性范围的新型l-酸盐生物传感器.
开发的生物传感器在未稀释的葡萄酒样本中准确量化l-酸盐,性能优于以前的方法.
这项工作为先进的电化学生物传感器铺平了道路,为食品和饮料应用提供了增强的功能.

关键词:

阿斯卡里斯 (Ascaris suum malic) 是一种有机酶.生物电子学生物电子学电化学生物传感器电子介质器是一种电子介质器.酶的表征酶的表征用丝网打印的电极.葡萄酒制作葡萄酒制作

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