A microfluidic bioreactor based on hydrogel-entrapped E. coli:: Cell viability, lysis, and intracellular enzyme reactions
A microfluidic bioreactor based on hydrogel-entrapped E. coli:: Cell viability, lysis, and intracellular enzyme reactions
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DOI：
10.1021/ac0259717
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发表时间：
2003-01-01
影响因子：
7.4
通讯作者：
Crooks, RM
中科院分区：
文献类型：
--
作者：
Heo, J;Thomas, KJ;Crooks, RM
活菌E.将大肠杆菌细胞包埋在微流体系统内光聚合的水凝胶微片中。微流体通道和微贴片具有大约100-500 μ m的尺寸。水凝胶周围的溶液中存在的小分子，如染料和表面活性剂，能够扩散到凝胶中并遇到细胞，但细胞足够大以被保留。例如，十二烷基硫酸钠是能够穿透水凝胶并破坏细胞膜的裂解剂。将活细胞包埋在水凝胶中，然后裂解，可以提供一种方便的方法来制备生物催化剂，而不需要酶的提取和纯化。水凝胶固定化细胞能够在微流体通道内进行化学反应。具体而言，非荧光染料BCECF-AM能够穿透水凝胶和细菌膜，并通过内部细胞机制转化为荧光形式（BCECF）。这些结果表明，固定在微流体通道内的细胞可以作为小分子的传感器和进行反应的生物反应器。
Viable E. coli cells were entrapped in hydrogel micropatches photopolymerized within microfluidic systems. The microfluidic channels and the micropatches have sizes on the order of 100-500 mum. Small molecules, such as dyes and surfactants, present in the solution surrounding the hydrogel, are able to diffuse into the gel and encounter the cells, but the cells are sufficiently large to be retained. For example, sodium dodecyl sulfate is a lysis agent that is able to penetrate the hydrogel and disrupt the cellular membrane. Entrapment of viable cells within hydrogels, followed by lysis, could provide a convenient means for preparing biocatalysts without the need for enzyme extraction and purification. Hydrogel-immobilized cells are able to carry out chemical reactions within microfluidic channels. Specifically, a nonfluorescent dye, BCECF-AM, is able to penetrate both the hydrogel and the bacterial membrane and be converted into a fluorescent form (BCECF) by the interior cellular machinery. These results suggest that cells immobilized within microfluidic channels can act as sensors for small molecules and as bioreactors for carrying out reactions.