Recombinant expression and purification of squid suckerin-spider silk fusion proteins for formation of thermosensitive hydrogel and biomimetic production of silk fibers

The suckerin from giant squid Dosidicus gigas and the spider silk are both protein-based biomaterials with remarkable mechanical properties, and thus they have great potential for the engineering of new biomimetic materials, such as fibers and thermosensitive hydrogels for biomedical applications. H...

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Main Author: Koh, Kenrick Yee Hong
Other Authors: Ali Gilles Tchenguise Miserez
Format: Final Year Project
Language:English
Published: 2018
Subjects:
Online Access:http://hdl.handle.net/10356/73767
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-737672023-03-04T15:36:54Z Recombinant expression and purification of squid suckerin-spider silk fusion proteins for formation of thermosensitive hydrogel and biomimetic production of silk fibers Koh, Kenrick Yee Hong Ali Gilles Tchenguise Miserez School of Materials Science and Engineering DRNTU::Engineering::Materials The suckerin from giant squid Dosidicus gigas and the spider silk are both protein-based biomaterials with remarkable mechanical properties, and thus they have great potential for the engineering of new biomimetic materials, such as fibers and thermosensitive hydrogels for biomedical applications. However, development of these applications requires robust recombinant expression systems that allow for production of high amount of spider silk and suckerin proteins. So far, efficient recombinant production of these proteins has been difficult to achieve due to their primary sequence characteristics i.e. high hydrophobicity and modular repeats that decrease their solubility. Recent studies have showed that these issues can be overcome by using a chimeric spider silk protein and shortening the repetitive sequences. Using this approach, we designed novel spider silk-squid suckerin fusion proteins. We successfully expressed and purified these proteins, and achieved a high production-yield of up to 154mg/L. Moreover, we demonstrated that the fusion proteins are capable of thermosensitive gelation to form hydrogels with tunable properties, making them an attractive application for biomedical scaffolds. We also attempted to biomimetically create mechanically stronger fibers using the fusion proteins. Bachelor of Engineering (Materials Engineering) 2018-04-09T05:12:30Z 2018-04-09T05:12:30Z 2018 Final Year Project (FYP) http://hdl.handle.net/10356/73767 en Nanyang Technological University 59 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::Engineering::Materials
spellingShingle DRNTU::Engineering::Materials
Koh, Kenrick Yee Hong
Recombinant expression and purification of squid suckerin-spider silk fusion proteins for formation of thermosensitive hydrogel and biomimetic production of silk fibers
description The suckerin from giant squid Dosidicus gigas and the spider silk are both protein-based biomaterials with remarkable mechanical properties, and thus they have great potential for the engineering of new biomimetic materials, such as fibers and thermosensitive hydrogels for biomedical applications. However, development of these applications requires robust recombinant expression systems that allow for production of high amount of spider silk and suckerin proteins. So far, efficient recombinant production of these proteins has been difficult to achieve due to their primary sequence characteristics i.e. high hydrophobicity and modular repeats that decrease their solubility. Recent studies have showed that these issues can be overcome by using a chimeric spider silk protein and shortening the repetitive sequences. Using this approach, we designed novel spider silk-squid suckerin fusion proteins. We successfully expressed and purified these proteins, and achieved a high production-yield of up to 154mg/L. Moreover, we demonstrated that the fusion proteins are capable of thermosensitive gelation to form hydrogels with tunable properties, making them an attractive application for biomedical scaffolds. We also attempted to biomimetically create mechanically stronger fibers using the fusion proteins.
author2 Ali Gilles Tchenguise Miserez
author_facet Ali Gilles Tchenguise Miserez
Koh, Kenrick Yee Hong
format Final Year Project
author Koh, Kenrick Yee Hong
author_sort Koh, Kenrick Yee Hong
title Recombinant expression and purification of squid suckerin-spider silk fusion proteins for formation of thermosensitive hydrogel and biomimetic production of silk fibers
title_short Recombinant expression and purification of squid suckerin-spider silk fusion proteins for formation of thermosensitive hydrogel and biomimetic production of silk fibers
title_full Recombinant expression and purification of squid suckerin-spider silk fusion proteins for formation of thermosensitive hydrogel and biomimetic production of silk fibers
title_fullStr Recombinant expression and purification of squid suckerin-spider silk fusion proteins for formation of thermosensitive hydrogel and biomimetic production of silk fibers
title_full_unstemmed Recombinant expression and purification of squid suckerin-spider silk fusion proteins for formation of thermosensitive hydrogel and biomimetic production of silk fibers
title_sort recombinant expression and purification of squid suckerin-spider silk fusion proteins for formation of thermosensitive hydrogel and biomimetic production of silk fibers
publishDate 2018
url http://hdl.handle.net/10356/73767
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