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基于蓝藻藻蓝蛋白的电化学生物传感器用于检测自由基过氧化氢

Cyanobacterial Phycocyanin-Based Electrochemical Biosensor for the Detection of the Free Radical Hydrogen Peroxide.

作者信息

Sivasankari Sivaprakasam, Vinoth Mani, Ravindran Abraham David

机构信息

Department of Microbiology, Periyar Arts College, Annamalai University, 607 001, Chidambaram, TN, India.

Department of Botany, College of Arts and Science, Periyar University, Sri Vijay Vidyalaya, Dharmapuri, 636 807, TN, India.

出版信息

Appl Biochem Biotechnol. 2024 Dec;196(12):8907-8924. doi: 10.1007/s12010-024-04978-4. Epub 2024 Jul 25.

Abstract

Cyanobacteria are photosynthetic prokaryotes that inhabit extreme environments by modifying their photosensitive chemoreceptors called cyanobacteriochromes (CBCRs) which are linear tetrapyrrole-linked phycobilin molecules. These light-sensitive phycobilin from Spirulina platensis is recognized as a potential photoreceptor tool in optogenetics for monitoring cellular morphogenesis. We prepared and extracted highly fluorescent cyanobacterial phycocyanin (C-PC) by irradiating the culture with ambient red light. The crude phycocyanin was subjected to ion exchange chromatography, and its purity was monitored using UV-visible, fluorescence, and FT-IR spectroscopy methods. In the conventional method, red light-induced C-PC exhibited strong antioxidant activity against HO, with 88.7% in vitro scavenging activity without requiring any other preservatives. Interestingly, this red light-acclimated phycocyanin was applied as a biosensing material for the detection of the free radical hydrogen peroxide (HO) using the enzyme horseradish peroxidase (HRP) as a mediator. The modified C-PC-HRP glassy carbon electrode (GCE) can detect HO from 0.1 to 1600 µM. The lowest possible detection limit of the electrode for HO was 19 nM. This electrode was used to detect free radical HO in blood serum samples. The microstructure of the lyophilized PC under SEM showed a flat crystal pattern, which enabled the immobilization of HRP on the electrode surface and electron transfer.

摘要

蓝藻是光合原核生物,它们通过修饰称为蓝藻光色素(CBCRs)的光敏化学感受器来栖息于极端环境,这些感受器是线性四吡咯连接的藻胆素分子。来自钝顶螺旋藻的这种光敏藻胆素被认为是光遗传学中用于监测细胞形态发生的一种潜在光感受器工具。我们通过用环境红光照射培养物来制备并提取高荧光蓝藻藻蓝蛋白(C-PC)。将粗藻蓝蛋白进行离子交换色谱分析,并使用紫外可见光谱、荧光光谱和傅里叶变换红外光谱方法监测其纯度。在传统方法中,红光诱导的C-PC对羟基自由基(HO)表现出很强的抗氧化活性,体外清除活性为88.7%,且无需任何其他防腐剂。有趣的是,这种经红光适应的藻蓝蛋白被用作生物传感材料,以辣根过氧化物酶(HRP)作为介质来检测自由基过氧化氢(HO)。修饰后的C-PC-HRP玻碳电极(GCE)能够检测0.1至1600 μM 的HO。该电极对HO的最低检测限为19 nM。此电极用于检测血清样本中的自由基HO。扫描电子显微镜下冻干藻蓝蛋白的微观结构显示出扁平的晶体图案,这使得HRP能够固定在电极表面并实现电子转移。

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