Halogenation of Peptides and Proteins Using Engineered Tryptophan Halogenase Enzymes
Halogenation of bioactive peptides via incorporation of non-natural amino acid derivatives during chemical synthesis is a common strategy to enhance functionality. Bacterial tyrptophan halogenases efficiently catalyze regiospecific halogenation of the free amino acid tryptophan, both in vitro and in...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-12-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/12/12/1841 |
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| author | Barindra Sana Ding Ke Eunice Hui Yen Li Timothy Ho Jayasree Seayad Hung A. Duong Farid J. Ghadessy |
| author_facet | Barindra Sana Ding Ke Eunice Hui Yen Li Timothy Ho Jayasree Seayad Hung A. Duong Farid J. Ghadessy |
| author_sort | Barindra Sana |
| collection | DOAJ |
| description | Halogenation of bioactive peptides via incorporation of non-natural amino acid derivatives during chemical synthesis is a common strategy to enhance functionality. Bacterial tyrptophan halogenases efficiently catalyze regiospecific halogenation of the free amino acid tryptophan, both in vitro and in vivo. Expansion of their substrate scope to peptides and proteins would facilitate highly-regulated post-synthesis/expression halogenation. Here, we demonstrate novel in vitro halogenation (chlorination and bromination) of peptides by select halogenase enzymes and identify the C-terminal (G/S)GW motif as a preferred substrate. In a first proof-of-principle experiment, we also demonstrate chemo-catalyzed derivatization of an enzymatically chlorinated peptide, albeit with low efficiency. We further rationally derive PyrH halogenase mutants showing improved halogenation of the (G/S)GW motif, both as a free peptide and when genetically fused to model proteins with efficiencies up to 90%. |
| first_indexed | 2024-03-09T17:16:47Z |
| format | Article |
| id | doaj.art-f48b338e0b4a4a07b8e703bd51df862a |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-09T17:16:47Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-f48b338e0b4a4a07b8e703bd51df862a2023-11-24T13:34:18ZengMDPI AGBiomolecules2218-273X2022-12-011212184110.3390/biom12121841Halogenation of Peptides and Proteins Using Engineered Tryptophan Halogenase EnzymesBarindra Sana0Ding Ke1Eunice Hui Yen Li2Timothy Ho3Jayasree Seayad4Hung A. Duong5Farid J. Ghadessy6Disease Intervention Technology Laboratory, Institute of Molecular and Cellular Biology, Agency for Science, Technology and Research (A*STAR), 8A Biomedical Grove, #06-04/05 Neuros/Immunos, Singapore 138648, SingaporeDisease Intervention Technology Laboratory, Institute of Molecular and Cellular Biology, Agency for Science, Technology and Research (A*STAR), 8A Biomedical Grove, #06-04/05 Neuros/Immunos, Singapore 138648, SingaporeInstitute of Sustainability for Chemicals, Energy and Environment, A*STAR, 8 Biomedical Grove, Neuros, #07-01, Singapore 138665, SingaporeInstitute of Sustainability for Chemicals, Energy and Environment, A*STAR, 8 Biomedical Grove, Neuros, #07-01, Singapore 138665, SingaporeInstitute of Sustainability for Chemicals, Energy and Environment, A*STAR, 8 Biomedical Grove, Neuros, #07-01, Singapore 138665, SingaporeInstitute of Sustainability for Chemicals, Energy and Environment, A*STAR, 8 Biomedical Grove, Neuros, #07-01, Singapore 138665, SingaporeDisease Intervention Technology Laboratory, Institute of Molecular and Cellular Biology, Agency for Science, Technology and Research (A*STAR), 8A Biomedical Grove, #06-04/05 Neuros/Immunos, Singapore 138648, SingaporeHalogenation of bioactive peptides via incorporation of non-natural amino acid derivatives during chemical synthesis is a common strategy to enhance functionality. Bacterial tyrptophan halogenases efficiently catalyze regiospecific halogenation of the free amino acid tryptophan, both in vitro and in vivo. Expansion of their substrate scope to peptides and proteins would facilitate highly-regulated post-synthesis/expression halogenation. Here, we demonstrate novel in vitro halogenation (chlorination and bromination) of peptides by select halogenase enzymes and identify the C-terminal (G/S)GW motif as a preferred substrate. In a first proof-of-principle experiment, we also demonstrate chemo-catalyzed derivatization of an enzymatically chlorinated peptide, albeit with low efficiency. We further rationally derive PyrH halogenase mutants showing improved halogenation of the (G/S)GW motif, both as a free peptide and when genetically fused to model proteins with efficiencies up to 90%.https://www.mdpi.com/2218-273X/12/12/1841halogenationproteinpeptidePyrHenzyme engineering |
| spellingShingle | Barindra Sana Ding Ke Eunice Hui Yen Li Timothy Ho Jayasree Seayad Hung A. Duong Farid J. Ghadessy Halogenation of Peptides and Proteins Using Engineered Tryptophan Halogenase Enzymes Biomolecules halogenation protein peptide PyrH enzyme engineering |
| title | Halogenation of Peptides and Proteins Using Engineered Tryptophan Halogenase Enzymes |
| title_full | Halogenation of Peptides and Proteins Using Engineered Tryptophan Halogenase Enzymes |
| title_fullStr | Halogenation of Peptides and Proteins Using Engineered Tryptophan Halogenase Enzymes |
| title_full_unstemmed | Halogenation of Peptides and Proteins Using Engineered Tryptophan Halogenase Enzymes |
| title_short | Halogenation of Peptides and Proteins Using Engineered Tryptophan Halogenase Enzymes |
| title_sort | halogenation of peptides and proteins using engineered tryptophan halogenase enzymes |
| topic | halogenation protein peptide PyrH enzyme engineering |
| url | https://www.mdpi.com/2218-273X/12/12/1841 |
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