Recommanded Product: 96-13-9. The reaction of aromatic heterocyclic molecules with protons is called protonation. Aromatic heterocycles are more basic than benzene due to the participation of heteroatoms. Compound: 2,3-Dibromo-1-propanol, is researched, Molecular C3H6Br2O, CAS is 96-13-9, about A paper-based whole-cell screening assay for directed evolution-driven enzyme engineering. Author is Gul, Ijaz; Bogale, Tadesse Fantaye; Chen, Yong; Yang, Xian; Fang, Ruiqin; Feng, Juan; Gao, Hui; Tang, Lixia.
However, the need for a large reagent volume and sophisticated instrumentation hampers its broad implementation. In an attempt to address this problem, here we report a paper-based high-throughput screening approach that should find broad application in generating desired enzymes. As an example case, the dehalogenation reaction of the halohydrin dehalogenase was adopted for assay development. In addition to visual detection, quant. measurements were performed by measuring the color intensity of an image that was photographed by a smartphone and processed using ImageJ free software. The proposed method was first validated using a gold standard method and then applied to mutagenesis library screening with reduced consumption of reagents (i.e., = 10μl per assay) and a shorter assay time. We identified two active mutants (P135A and G137A) with improved activities toward four tested substrates. The assay not only consumes less reagents but also eliminates the need for expensive instrumentation. The proposed method demonstrates the potential of paper-based whole-cell screening coupled with digital image colorimetry as a promising approach for the discovery of industrially important enzymes. Key Points · A frugal method was developed for directed enzyme evolution. · Mutagenesis libraries were successfully screened on a paper platform. · Smartphone imaging was efficiently used to measure enzyme activities.
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