MilliporeSigma

Mapping enzyme catalysis with metabolic biosensing.

Nature communications (2021-11-25)
Linfeng Xu, Kai-Chun Chang, Emory M Payne, Cyrus Modavi, Leqian Liu, Claire M Palmer, Nannan Tao, Hal S Alper, Robert T Kennedy, Dale S Cornett, Adam R Abate
ABSTRACT

Enzymes are represented across a vast space of protein sequences and structural forms and have activities that far exceed the best chemical catalysts; however, engineering them to have novel or enhanced activity is limited by technologies for sensing product formation. Here, we describe a general and scalable approach for characterizing enzyme activity that uses the metabolism of the host cell as a biosensor by which to infer product formation. Since different products consume different molecules in their synthesis, they perturb host metabolism in unique ways that can be measured by mass spectrometry. This provides a general way by which to sense product formation, to discover unexpected products and map the effects of mutagenesis.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
LB Agar Ampicillin-100, Plates, pre-poured LB agar plates with 100 μg/ml ampicillin