期刊
FRONTIERS IN MICROBIOLOGY
卷 11, 期 -, 页码 -出版社
FRONTIERS MEDIA SA
DOI: 10.3389/fmicb.2020.00760
关键词
Pseudomonas aeruginosa; biofilm; PslG; trypsin; protease
类别
资金
- National Natural Science Foundation of China [31470732]
A glycosyl hydrolase produced by Pseudomonas aeruginosa, PslG, has become a promising candidate for biofilm treatment because of its ability to inhibit and disperse biofilms by disrupting exopolysaccharide matrix at nanomolar concentrations. However, as a protein, PslG used for treatment may be degraded by the ubiquitous proteases (of which trypsin-like serine proteases are a major group) secreted by human cells. This would lead to an insufficient effective concentration of PslG. Here, based on the result of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and structural analysis, we generate a PslG mutant (K286A/K433S) with greatly enhanced trypsin resistance. This measure raises IC50 (the concentration of trypsin that can degrade 50% of protein in 30 min at 37 degrees C) from 0.028 mg mL(-1) of the wild-type PslG to 0.283 mg mL(-1) of PslG(K286A/K433S). In addition, biofilm inhibition assay shows that PslG(K286A/K433S) is much more efficient than wild-type PslG in the presence of trypsin. This indicates that PslG(K286A/K433S) is a better biofilm inhibitor than wild-type PslG in clinical use where trypsin-like proteases widely exist.
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