Yasaputera, S (2016) The role of Ser497 in Phosphopantetheinylationof Tcp11, an LHydroxyphenylglycine activating module. Research Project 1 (minor thesis), Molecular Biology and Biotechnology (2016-2019).
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Abstract
Nonribosomal peptides synthetases (NRPS) are multimodular biocatalysts that build diverse pool of biologically active natural compounds. Nonribosomal peptides (NRPs) are commonly used as antibiotics, antitumor, antifungal or as immunosuppressant. Within multimodular structure of NRPSs, each module comprises of several domains, such as adenylation (A), thiolation (PCP), condensation (C), and thioesterase (TE) domain. One of the important domains is thiolation (PCP) domain, as a strictly conserved serine residue (Ser497) acts in the active site to allow Ppant-arm formation. During this project, another two serine residues (Ser499 and Ser500), located downstream of the conserved Ser497 in PCP-domain of L-Hydroxyphenylglycine activating module (Tcp11) were included in the study. In order to investigate the role of Ser497, the neighboring Ser499 and Ser500 in Ppant-arm formation of Tcp11, a mutagenesis strategy was employed. Therefore single mutations were performed of the serine residues into alanine. Additionally, a triple serine knockout was prepared where all three serine residues were exchanged to alanine (S497A/S499A/S500A). Subsequently, the wild type and the mutants were expressed in E.coli and their adenylation and thiolation activity were tested.
| Item Type: | Thesis (Research Project 1 (minor thesis)) |
|---|---|
| Degree programme: | Molecular Biology and Biotechnology (2016-2019) |
| Thesis type: | Research Project 1 (minor thesis) |
| Language: | English |
| Date Deposited: | 15 Feb 2018 08:26 |
| Last Modified: | 15 Feb 2018 08:26 |
| URI: | https://fse.studenttheses.ub.rug.nl/id/eprint/14736 |
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