Towards a general approach for tailoring the hydrophobic binding site of phenylalanine ammonia-lyases
Abstract Unnatural substituted amino acids play an important role as chiral building blocks, especially for pharmaceutical industry, where the synthesis of chiral biologically active molecules still represents an open challenge. Recently, modification of the hydrophobic binding pocket of phenylalani...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-06-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-14585-0 |
| _version_ | 1818249850815250432 |
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| author | Souad Diana Tork Mădălina Elena Moisă Lilla Cserepes Alina Filip Levente Csaba Nagy Florin Dan Irimie László Csaba Bencze |
| author_facet | Souad Diana Tork Mădălina Elena Moisă Lilla Cserepes Alina Filip Levente Csaba Nagy Florin Dan Irimie László Csaba Bencze |
| author_sort | Souad Diana Tork |
| collection | DOAJ |
| description | Abstract Unnatural substituted amino acids play an important role as chiral building blocks, especially for pharmaceutical industry, where the synthesis of chiral biologically active molecules still represents an open challenge. Recently, modification of the hydrophobic binding pocket of phenylalanine ammonia-lyase from Petroselinum crispum (PcPAL) resulted in specifically tailored PcPAL variants, contributing to a rational design template for PAL-activity enhancements towards the differently substituted substrate analogues. Within this study we tested the general applicability of this rational design model in case of PALs, of different sources, such as from Arabidopsis thaliana (AtPAL) and Rhodosporidium toruloides (RtPAL). With some exceptions, the results support that the positions of substrate specificity modulating residues are conserved among PALs, thus the mutation with beneficial effect for PAL-activity enhancement can be predicted using the established rational design model. Accordingly, the study supports that tailoring PALs of different origins and different substrate scope, can be performed through a general method. Moreover, the fact that AtPAL variants I461V, L133A and L257V, all outperformed in terms of catalytic efficiency the corresponding, previously reported, highly efficient PcPAL variants, of identical catalytic site, suggests that not only catalytic site differences influence the PAL-activity, thus for the selection of the optimal PAL-biocatalysts for a targeted process, screening of PALs from different origins, should be included. |
| first_indexed | 2024-12-12T15:43:03Z |
| format | Article |
| id | doaj.art-a1ac952428c94f1ea4e20017a24a8f9e |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-12T15:43:03Z |
| publishDate | 2022-06-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-a1ac952428c94f1ea4e20017a24a8f9e2022-12-22T00:19:51ZengNature PortfolioScientific Reports2045-23222022-06-0112111210.1038/s41598-022-14585-0Towards a general approach for tailoring the hydrophobic binding site of phenylalanine ammonia-lyasesSouad Diana Tork0Mădălina Elena Moisă1Lilla Cserepes2Alina Filip3Levente Csaba Nagy4Florin Dan Irimie5László Csaba Bencze6Enzymology and Applied Biocatalysis Research Center, Faculty of Chemistry and Chemical Engineering, Babeș-Bolyai UniversityEnzymology and Applied Biocatalysis Research Center, Faculty of Chemistry and Chemical Engineering, Babeș-Bolyai UniversityEnzymology and Applied Biocatalysis Research Center, Faculty of Chemistry and Chemical Engineering, Babeș-Bolyai UniversityEnzymology and Applied Biocatalysis Research Center, Faculty of Chemistry and Chemical Engineering, Babeș-Bolyai UniversityEnzymology and Applied Biocatalysis Research Center, Faculty of Chemistry and Chemical Engineering, Babeș-Bolyai UniversityEnzymology and Applied Biocatalysis Research Center, Faculty of Chemistry and Chemical Engineering, Babeș-Bolyai UniversityEnzymology and Applied Biocatalysis Research Center, Faculty of Chemistry and Chemical Engineering, Babeș-Bolyai UniversityAbstract Unnatural substituted amino acids play an important role as chiral building blocks, especially for pharmaceutical industry, where the synthesis of chiral biologically active molecules still represents an open challenge. Recently, modification of the hydrophobic binding pocket of phenylalanine ammonia-lyase from Petroselinum crispum (PcPAL) resulted in specifically tailored PcPAL variants, contributing to a rational design template for PAL-activity enhancements towards the differently substituted substrate analogues. Within this study we tested the general applicability of this rational design model in case of PALs, of different sources, such as from Arabidopsis thaliana (AtPAL) and Rhodosporidium toruloides (RtPAL). With some exceptions, the results support that the positions of substrate specificity modulating residues are conserved among PALs, thus the mutation with beneficial effect for PAL-activity enhancement can be predicted using the established rational design model. Accordingly, the study supports that tailoring PALs of different origins and different substrate scope, can be performed through a general method. Moreover, the fact that AtPAL variants I461V, L133A and L257V, all outperformed in terms of catalytic efficiency the corresponding, previously reported, highly efficient PcPAL variants, of identical catalytic site, suggests that not only catalytic site differences influence the PAL-activity, thus for the selection of the optimal PAL-biocatalysts for a targeted process, screening of PALs from different origins, should be included.https://doi.org/10.1038/s41598-022-14585-0 |
| spellingShingle | Souad Diana Tork Mădălina Elena Moisă Lilla Cserepes Alina Filip Levente Csaba Nagy Florin Dan Irimie László Csaba Bencze Towards a general approach for tailoring the hydrophobic binding site of phenylalanine ammonia-lyases Scientific Reports |
| title | Towards a general approach for tailoring the hydrophobic binding site of phenylalanine ammonia-lyases |
| title_full | Towards a general approach for tailoring the hydrophobic binding site of phenylalanine ammonia-lyases |
| title_fullStr | Towards a general approach for tailoring the hydrophobic binding site of phenylalanine ammonia-lyases |
| title_full_unstemmed | Towards a general approach for tailoring the hydrophobic binding site of phenylalanine ammonia-lyases |
| title_short | Towards a general approach for tailoring the hydrophobic binding site of phenylalanine ammonia-lyases |
| title_sort | towards a general approach for tailoring the hydrophobic binding site of phenylalanine ammonia lyases |
| url | https://doi.org/10.1038/s41598-022-14585-0 |
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