Jung Sukwon, MacConaghy Kelsey I, Guarnieri Michael T, Kaar Joel L, Stoykovich Mark P
Department of Chemical and Biological Engineering, University of Colorado, Boulder, Colorado 80303, United States.
Biosciences Center, National Renewable Energy Laboratory, Golden, Colorado 80401, United States.
ACS Appl Bio Mater. 2022 Mar 21;5(3):1252-1258. doi: 10.1021/acsabm.1c01267. Epub 2022 Feb 15.
We herein describe a highly versatile platform approach for the in situ and real-time screening of microbial biocatalysts for enhanced production of bioproducts using photonic crystal hydrogels. This approach was demonstrated by preparing optically diffracting films based on polymerized -isopropylacrylamide that contracted in the presence of alcohols and organic acids. The hydrogel films were prepared in a microwell plate format, which allows for high-throughput screening, and characterized optically using a microwell plate reader. While demonstrating the ability to detect a broad range of relevant alcohols and organic acids, we showed that the response of the films correlated strongly with the octanol-water partition coefficient (log ) of the analyte. Differences in the secretion of ethanol and succinic acid from strains of and , respectively, were further detected via optical characterization of the films. These differences, which in some cases were as low as ∼3 g/L, were confirmed by high-performance liquid chromatography, thereby demonstrating the sensitivity of this approach. Our findings highlight the potential utility of this multiplexed approach for the detection of small organic analytes in complex biological media, which overcomes a major challenge in conventional optical sensing methods.
我们在此描述了一种高度通用的平台方法,用于使用光子晶体水凝胶对微生物生物催化剂进行原位实时筛选,以提高生物产品的产量。通过制备基于聚合异丙基丙烯酰胺的光学衍射膜来证明这种方法,该膜在醇和有机酸存在下会收缩。水凝胶膜以微孔板形式制备,允许高通量筛选,并使用微孔板读数器进行光学表征。在展示检测多种相关醇和有机酸的能力时,我们表明膜的响应与分析物的辛醇 - 水分配系数(log )密切相关。通过膜的光学表征进一步检测了分别来自 和 菌株的乙醇和琥珀酸分泌差异。这些差异在某些情况下低至约3 g/L,通过高效液相色谱法得到证实,从而证明了该方法的灵敏度。我们的研究结果突出了这种多重方法在检测复杂生物介质中小有机分析物方面的潜在效用,这克服了传统光学传感方法中的一个主要挑战。