Screening method of bacterial enoyl acyl carrier protein (ACP) reductase inhibitor

Abstract

The invention discloses a screening method of a bacterial enoyl acyl carrier protein (ACP) reductase inhibitor. The screening method comprises the steps: expressing fabK of enterococcus faecalis in escherichia coli, inhibiting and not inhibiting FabI of recombinant escherichia coli through triclosan, and constructing a double-plate sensitive difference screening model for FabK; constructing a fabI gene deletion mutant and a fabV gene deletion mutant of pseudomonas aeruginosa, and constructing a three-plate sensitive difference screening model for FabI and FabV on the basis of wild bacteria; screening a combination of the two screening models, and measuring half inhibition concentration of an activity extract on enoyl ACP reductase and detecting different expression strains of the enoyl ACP reductase to the activity extract, thus realizing secondary screening on sensitivity difference of the activity extract. The screening method disclosed by the invention has the beneficial effects that the screening method not only improves screening efficiency and simplifies screening of a broad-spectrum inhibitor, but also avoids construction of various drug screening models for diversity of the enoyl ACP reductase; and simultaneously, the method solves problems of high false positive and low screening efficiency.

Description

technical field [0001] The invention relates to a screening method for inhibitors, in particular to a screening method for bacterial enoyl ACP reductase inhibitors, and belongs to the technical field of molecular biology. Background technique [0002] Because bacteria have developed resistance to existing antibiotics, they still seriously threaten human health, and even super-resistant bacteria that have caused global panic have emerged. Although the scientific community has been very enthusiastic about finding new antibiotics that act on new targets, these efforts have had limited results since the 1960s. Although hundreds of essential bacterial proteins have been identified as potential drug targets, only a few of them have become targets for clinical therapeutics. Over the past few decades, effective antibiotics have been developed by chemically modifying the backbone of existing antibiotics to enhance their activity; however, it has become increasingly difficult to deve...

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Problems solved by technology

However, the screening efficiency of this method is relatively low

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