Butelase 1 : the fastest known ligase for protein cyclization and ligation

Peptide cyclization confers increased conformational stability and rigidity against metabolic and thermal denaturation. Encouraged by potential usage of cyclic peptides in the therapeutic applications, methods of protein cyclization and ligation have been developed such as native chemical ligation,...

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Main Author: Chen, Shu Bee
Other Authors: Jimmy Pingkwan Tam @ James P Tam
Format: Final Year Project
Language:English
Published: 2014
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Online Access:http://hdl.handle.net/10356/61748
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-617482023-02-28T18:05:47Z Butelase 1 : the fastest known ligase for protein cyclization and ligation Chen, Shu Bee Jimmy Pingkwan Tam @ James P Tam School of Biological Sciences DRNTU::Science Peptide cyclization confers increased conformational stability and rigidity against metabolic and thermal denaturation. Encouraged by potential usage of cyclic peptides in the therapeutic applications, methods of protein cyclization and ligation have been developed such as native chemical ligation, intein-, PatG- and sortase A- mediated transpeptidase activity. However, with various kinds of limitations in restricting the practical applications, a natural occurring ligase, butelase 1 has been discovered and isolated from Clitoria ternatea recently. Here, I perform the enzyme kinetics characterization of butelase 1 for cyclization of various peptides of plant and animal origin and explore the potential applications in mediating intermolecular ligation. With the kcat value up to 4 s-1 and the catalytic efficiency of 71,384 M-1s-1, butelase 1 is the fastest known ligase. Butelase 1, an Asn-specific ligase, cyclizes various bioactive peptides quantitatively in a head-to-tail manner with conversion yield >95% and displays broad substrate specificity. Moreover, it is useful for introducing a functional group such as biotin and fluorophore to proteins by using GFP, monoclonal antibody and darpin as model proteins. The results show that butelase 1 would play an important role in the future bioengineering and pharmaceutical applications. Bachelor of Science in Biomedical Sciences 2014-09-12T02:51:04Z 2014-09-12T02:51:04Z 2014 2014 Final Year Project (FYP) http://hdl.handle.net/10356/61748 en Nanyang Technological University 33 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science
spellingShingle DRNTU::Science
Chen, Shu Bee
Butelase 1 : the fastest known ligase for protein cyclization and ligation
description Peptide cyclization confers increased conformational stability and rigidity against metabolic and thermal denaturation. Encouraged by potential usage of cyclic peptides in the therapeutic applications, methods of protein cyclization and ligation have been developed such as native chemical ligation, intein-, PatG- and sortase A- mediated transpeptidase activity. However, with various kinds of limitations in restricting the practical applications, a natural occurring ligase, butelase 1 has been discovered and isolated from Clitoria ternatea recently. Here, I perform the enzyme kinetics characterization of butelase 1 for cyclization of various peptides of plant and animal origin and explore the potential applications in mediating intermolecular ligation. With the kcat value up to 4 s-1 and the catalytic efficiency of 71,384 M-1s-1, butelase 1 is the fastest known ligase. Butelase 1, an Asn-specific ligase, cyclizes various bioactive peptides quantitatively in a head-to-tail manner with conversion yield >95% and displays broad substrate specificity. Moreover, it is useful for introducing a functional group such as biotin and fluorophore to proteins by using GFP, monoclonal antibody and darpin as model proteins. The results show that butelase 1 would play an important role in the future bioengineering and pharmaceutical applications.
author2 Jimmy Pingkwan Tam @ James P Tam
author_facet Jimmy Pingkwan Tam @ James P Tam
Chen, Shu Bee
format Final Year Project
author Chen, Shu Bee
author_sort Chen, Shu Bee
title Butelase 1 : the fastest known ligase for protein cyclization and ligation
title_short Butelase 1 : the fastest known ligase for protein cyclization and ligation
title_full Butelase 1 : the fastest known ligase for protein cyclization and ligation
title_fullStr Butelase 1 : the fastest known ligase for protein cyclization and ligation
title_full_unstemmed Butelase 1 : the fastest known ligase for protein cyclization and ligation
title_sort butelase 1 : the fastest known ligase for protein cyclization and ligation
publishDate 2014
url http://hdl.handle.net/10356/61748
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