Rusling James F, Hvastkovs Eli G, Hull Dominic O, Schenkman John B
Department of Chemistry, University of Connecticut, Storrs, CT 06269, USA.
Chem Commun (Camb). 2008 Jan 14(2):141-54. doi: 10.1039/b709121b. Epub 2007 Aug 30.
This feature article summarizes recent applications of ultrathin films of enzymes and DNA assembled layer-by-layer (LbL). Using examples mainly from our own research, we focus on systems developed for biocatalysis and biosensors for toxicity screening. Enzyme-poly(L-lysine) (PLL) films, especially when stabilized by crosslinking, can be used for biocatalysis at unprecedented high temperatures or in acidic or basic solutions on electrodes or sub-micron sized beads. Such films have bright prospects for chiral synthesis and biofuel cells. Excellent bioactivity and retention of enzyme structure in these films facilitates their use in detailed kinetic studies. Biosensors and arrays employing DNA-enzyme films show great promise in predicting genotoxicity of new drug and chemical product candidates. These devices combine metabolic biocatalysis, reactive metabolite-DNA reactions, and DNA damage detection. Catalytic voltammetry or electrochemiluminescence (ECL) can be used for high throughput arrays utilizing multiple LbL "spots" of DNA, enzyme and metallopolymer. DNA-enzyme films can also be used to produce nucleobase adduct toxicity biomarkers for detection by LC-MS. These approaches provide valuable high throughput tools for drug and chemical product development and toxicity prediction.
这篇专题文章总结了酶和DNA逐层组装(LbL)超薄膜的近期应用。主要以我们自己的研究为例,我们重点关注为生物催化和毒性筛选生物传感器而开发的系统。酶-聚(L-赖氨酸)(PLL)薄膜,特别是通过交联稳定化后,可用于在前所未有的高温下或在电极或亚微米尺寸珠子上的酸性或碱性溶液中进行生物催化。此类薄膜在手性合成和生物燃料电池方面具有光明前景。这些薄膜中优异的生物活性和酶结构的保留便于其用于详细的动力学研究。采用DNA-酶薄膜的生物传感器和阵列在预测新药和化学产品候选物的遗传毒性方面显示出巨大潜力。这些装置结合了代谢生物催化、反应性代谢物-DNA反应以及DNA损伤检测。催化伏安法或电化学发光(ECL)可用于利用DNA、酶和金属聚合物的多个LbL“斑点”的高通量阵列。DNA-酶薄膜还可用于生产用于液相色谱-质谱检测的核碱基加合物毒性生物标志物。这些方法为药物和化学产品开发及毒性预测提供了有价值的高通量工具。