Manchester Synthetic Biology Research Centre for Fine and Speciality Chemicals (SYNBIOCHEM), Manchester Institute of Biotechnology, School of Chemistry, The University of Manchester , 131 Princess Street, M1 7DN, Manchester, United Kingdom.
Waters Corp., Stamford Avenue, Altrincham Road, SK9 4AX, Wilmslow, United Kingdom.
J Am Chem Soc. 2017 Feb 1;139(4):1408-1411. doi: 10.1021/jacs.6b12165. Epub 2017 Jan 19.
Screening of bacterial colonies to identify new biocatalytic activities is a widely adopted tool in biotechnology, but is constrained by the requirements for colorimetric or tag-based detection methods. Herein we report a label-free screening platform for biotransformations in live colonies using desorption electrospray ionization coupled with ion mobility mass spectrometry imaging (DiBT-IMMS). The screening method is demonstrated for both ammonia lyases and P450 monooxygenases expressed within live bacterial colonies and is shown to enable multiplexing of enzyme variants and substrate libraries simultaneously.
筛选具有新型生物催化活性的细菌菌落是生物技术中广泛采用的一种工具，但受到比色或基于标签的检测方法的限制。本文报道了一种使用解吸电喷雾电离结合离子淌度质谱成像（DiBT-IMMS）对活菌落中的生物转化进行无标记筛选的平台。该筛选方法在活细菌菌落中表达的氨裂解酶和 P450 单加氧酶中得到了验证，并显示出能够同时对酶变体和底物文库进行多重化。
