A Rational Designed PslG With Normal Biofilm Hydrolysis and Enhanced Resistance to Trypsin-Like Protease Digestion
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 degr...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2020-05-01
|
| Series: | Frontiers in Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2020.00760/full |
| _version_ | 1818901855855443968 |
|---|---|
| author | Tiantian Su Jing He Ningna Li Shiheng Liu Sujuan Xu Lichuan Gu |
| author_facet | Tiantian Su Jing He Ningna Li Shiheng Liu Sujuan Xu Lichuan Gu |
| author_sort | Tiantian Su |
| collection | DOAJ |
| description | 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°C) from 0.028 mg mL–1 of the wild-type PslG to 0.283 mg mL–1 of PslGK286A/K433S. In addition, biofilm inhibition assay shows that PslGK286A/K433S is much more efficient than wild-type PslG in the presence of trypsin. This indicates that PslGK286A/K433S is a better biofilm inhibitor than wild-type PslG in clinical use where trypsin-like proteases widely exist. |
| first_indexed | 2024-12-19T20:26:23Z |
| format | Article |
| id | doaj.art-2e6f55f8278e4b1b9baf8654a055e2ae |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-19T20:26:23Z |
| publishDate | 2020-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-2e6f55f8278e4b1b9baf8654a055e2ae2022-12-21T20:06:49ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2020-05-011110.3389/fmicb.2020.00760517063A Rational Designed PslG With Normal Biofilm Hydrolysis and Enhanced Resistance to Trypsin-Like Protease DigestionTiantian SuJing HeNingna LiShiheng LiuSujuan XuLichuan GuA 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°C) from 0.028 mg mL–1 of the wild-type PslG to 0.283 mg mL–1 of PslGK286A/K433S. In addition, biofilm inhibition assay shows that PslGK286A/K433S is much more efficient than wild-type PslG in the presence of trypsin. This indicates that PslGK286A/K433S is a better biofilm inhibitor than wild-type PslG in clinical use where trypsin-like proteases widely exist.https://www.frontiersin.org/article/10.3389/fmicb.2020.00760/fullPseudomonas aeruginosabiofilmPslGtrypsinprotease |
| spellingShingle | Tiantian Su Jing He Ningna Li Shiheng Liu Sujuan Xu Lichuan Gu A Rational Designed PslG With Normal Biofilm Hydrolysis and Enhanced Resistance to Trypsin-Like Protease Digestion Frontiers in Microbiology Pseudomonas aeruginosa biofilm PslG trypsin protease |
| title | A Rational Designed PslG With Normal Biofilm Hydrolysis and Enhanced Resistance to Trypsin-Like Protease Digestion |
| title_full | A Rational Designed PslG With Normal Biofilm Hydrolysis and Enhanced Resistance to Trypsin-Like Protease Digestion |
| title_fullStr | A Rational Designed PslG With Normal Biofilm Hydrolysis and Enhanced Resistance to Trypsin-Like Protease Digestion |
| title_full_unstemmed | A Rational Designed PslG With Normal Biofilm Hydrolysis and Enhanced Resistance to Trypsin-Like Protease Digestion |
| title_short | A Rational Designed PslG With Normal Biofilm Hydrolysis and Enhanced Resistance to Trypsin-Like Protease Digestion |
| title_sort | rational designed pslg with normal biofilm hydrolysis and enhanced resistance to trypsin like protease digestion |
| topic | Pseudomonas aeruginosa biofilm PslG trypsin protease |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2020.00760/full |
| work_keys_str_mv | AT tiantiansu arationaldesignedpslgwithnormalbiofilmhydrolysisandenhancedresistancetotrypsinlikeproteasedigestion AT jinghe arationaldesignedpslgwithnormalbiofilmhydrolysisandenhancedresistancetotrypsinlikeproteasedigestion AT ningnali arationaldesignedpslgwithnormalbiofilmhydrolysisandenhancedresistancetotrypsinlikeproteasedigestion AT shihengliu arationaldesignedpslgwithnormalbiofilmhydrolysisandenhancedresistancetotrypsinlikeproteasedigestion AT sujuanxu arationaldesignedpslgwithnormalbiofilmhydrolysisandenhancedresistancetotrypsinlikeproteasedigestion AT lichuangu arationaldesignedpslgwithnormalbiofilmhydrolysisandenhancedresistancetotrypsinlikeproteasedigestion AT tiantiansu rationaldesignedpslgwithnormalbiofilmhydrolysisandenhancedresistancetotrypsinlikeproteasedigestion AT jinghe rationaldesignedpslgwithnormalbiofilmhydrolysisandenhancedresistancetotrypsinlikeproteasedigestion AT ningnali rationaldesignedpslgwithnormalbiofilmhydrolysisandenhancedresistancetotrypsinlikeproteasedigestion AT shihengliu rationaldesignedpslgwithnormalbiofilmhydrolysisandenhancedresistancetotrypsinlikeproteasedigestion AT sujuanxu rationaldesignedpslgwithnormalbiofilmhydrolysisandenhancedresistancetotrypsinlikeproteasedigestion AT lichuangu rationaldesignedpslgwithnormalbiofilmhydrolysisandenhancedresistancetotrypsinlikeproteasedigestion |