Academic Journal
Design of Trypanosoma rangeli sialidase mutants with improved trans-sialidase activity
| العنوان: | Design of Trypanosoma rangeli sialidase mutants with improved trans-sialidase activity |
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| المؤلفون: | Nyffenegger, Christian, Nordvang, Rune Thorbjørn, Jers, Carsten, Meyer, Anne S., Mikkelsen, Jørn Dalgaard |
| المصدر: | Nyffenegger , C , Nordvang , R T , Jers , C , Meyer , A S & Mikkelsen , J D 2017 , ' Design of Trypanosoma rangeli sialidase mutants with improved trans-sialidase activity ' , PLOS ONE , vol. 12 , no. 2 , e0171585 . https://doi.org/10.1371/journal.pone.0171585 |
| سنة النشر: | 2017 |
| المجموعة: | Technical University of Denmark: DTU Orbit / Danmarks Tekniske Universitet |
| الوصف: | A sialidase (EC 3.2.1.18) from the non-pathogenic Trypanosoma rangeli, TrSA, has been shown to exert trans-sialidase activity after mutation of five specific amino acids in the active site (M96V, A98P, S120Y, G249Y, Q284P) to form the so-called TrSA 5mut enzyme. By computational and hypothesis driven approaches additional mutations enhancing the trans-sialidase activity have been suggested. In the present work, we made a systematic combination of these mutations leading to seven new variants of the T. rangeli sialidase, having 6-16 targeted amino acid mutations. The resulting enzyme variants were analyzed via kinetics for their ability to carry out trans-sialidase reaction using CGMP and D-lactose as substrates. The sialidase variants with 15 and 16 mutations, respectively, exhibited significantly improved trans-sialidase activity for D-lactose sialylation. Our results corroborate, that computational studies of trans-glycosylation can be a valuable input in the design of novel trans-glycosidases, but also highlight the importance of experimental validation in order to assess the performance. In conclusion, two of the seven mutants displayed a dramatic switch in specificity from hydrolysis towards trans-sialylation and constitute the most potent trans-sialidase mutants of TrSA described in literature to date. |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| Relation: | https://orbit.dtu.dk/en/publications/dccb289b-eab2-46e4-974d-9f50151d5a58 |
| DOI: | 10.1371/journal.pone.0171585 |
| الاتاحة: | https://orbit.dtu.dk/en/publications/dccb289b-eab2-46e4-974d-9f50151d5a58 https://doi.org/10.1371/journal.pone.0171585 https://backend.orbit.dtu.dk/ws/files/128949524/pone.0171585.pdf |
| Rights: | info:eu-repo/semantics/openAccess |
| رقم الانضمام: | edsbas.E7D76B6C |
| قاعدة البيانات: | BASE |
| DOI: | 10.1371/journal.pone.0171585 |
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