Real-Time Screening of Biocatalysts in Live Bacterial Colonies
Real-Time Screening of Biocatalysts in Live Bacterial Colonies
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DOI：
10.1021/jacs.6b12165
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发表时间：
2017-02-01
影响因子：
15
通讯作者：
Barran, Perdita E.
中科院分区：
文献类型：
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作者：
Yan, Cunyu;Parmeggiani, Fabio;Barran, Perdita E.
筛选细菌菌落以鉴定新的生物催化活性是生物技术中广泛采用的工具，但受到比色法或基于标签的检测方法的要求的限制。在这里，我们报道了一种利用解吸电喷雾电离结合离子迁移率质谱成像(DiBT-IMMS)的无标记筛选平台，用于活菌落中的生物转化。对活细菌菌落中表达的氨裂解酶和P450单加氧酶的筛选方法进行了验证，并证明该方法能够同时对酶变体和底物文库进行多路复用。
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.