Selection of potentially optimized trypsin variant with a more hydrophobic cluster active site via a phage display approach

Protein optimization can generally be obtained by rational enzyme design and directed evolution. Phage display has been used as a method for directed evolution. Trypsin has been studied as a biocatalyst in peptide synthesis despite its usual function in hydrolyzing peptide bonds. The objective of th...

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Main Authors:	Abdul Kudos, Muhammad Badri, Gissel, Sabrina, Bordusa, Frank
Format:	Conference or Workshop Item
Language:	English English
Published:	2015
Subjects:	TP248.13 Biotechnology
Online Access:	http://irep.iium.edu.my/48397/ http://irep.iium.edu.my/48397/1/Badri%20KOP.pdf http://irep.iium.edu.my/48397/2/Poster_badri_ACB2015.pdf

id	iium-48397
recordtype	eprints
spelling	iium-483972017-06-01T08:12:33Z http://irep.iium.edu.my/48397/ Selection of potentially optimized trypsin variant with a more hydrophobic cluster active site via a phage display approach Abdul Kudos, Muhammad Badri Gissel, Sabrina Bordusa, Frank TP248.13 Biotechnology Protein optimization can generally be obtained by rational enzyme design and directed evolution. Phage display has been used as a method for directed evolution. Trypsin has been studied as a biocatalyst in peptide synthesis despite its usual function in hydrolyzing peptide bonds. The objective of this study is to identify trypsin variants with a more hydrophobic cluster around its active site, potentially yielding higher peptide synthesis products rather than its hydrolysis competitive counterpart. A trypsin variant library displayed by phages with randomization in 5 amino acids located around the active site of the trypsin variant K60E/N143H/E151H/D189K is selected based on a designed immobilization-elution-assay in regard the transamidation reaction model. A preselection stage of recombinant phages was done with MyCUT tag, binding to the immobilized anti-c-myc antibodies. 2 trypsin variants were successfully expressed, purified, characterized and sequenced. Both of them gave a higher accumulative hydrophobicity relative index. This study has shown that selection of a potentially optimized trypsin variant with a more hydrophobic cluster at active site to reduce the hydrolysis products is possible by using the phage display approach. 2015 Conference or Workshop Item NonPeerReviewed application/pdf en http://irep.iium.edu.my/48397/1/Badri%20KOP.pdf application/pdf en http://irep.iium.edu.my/48397/2/Poster_badri_ACB2015.pdf Abdul Kudos, Muhammad Badri and Gissel, Sabrina and Bordusa, Frank (2015) Selection of potentially optimized trypsin variant with a more hydrophobic cluster active site via a phage display approach. In: ASIAN Congress on Biotechnology 2015, 15-19 Nov 2015, Kuala Lumpur. (Unpublished)
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English English
topic	TP248.13 Biotechnology
spellingShingle	TP248.13 Biotechnology Abdul Kudos, Muhammad Badri Gissel, Sabrina Bordusa, Frank Selection of potentially optimized trypsin variant with a more hydrophobic cluster active site via a phage display approach
description	Protein optimization can generally be obtained by rational enzyme design and directed evolution. Phage display has been used as a method for directed evolution. Trypsin has been studied as a biocatalyst in peptide synthesis despite its usual function in hydrolyzing peptide bonds. The objective of this study is to identify trypsin variants with a more hydrophobic cluster around its active site, potentially yielding higher peptide synthesis products rather than its hydrolysis competitive counterpart. A trypsin variant library displayed by phages with randomization in 5 amino acids located around the active site of the trypsin variant K60E/N143H/E151H/D189K is selected based on a designed immobilization-elution-assay in regard the transamidation reaction model. A preselection stage of recombinant phages was done with MyCUT tag, binding to the immobilized anti-c-myc antibodies. 2 trypsin variants were successfully expressed, purified, characterized and sequenced. Both of them gave a higher accumulative hydrophobicity relative index. This study has shown that selection of a potentially optimized trypsin variant with a more hydrophobic cluster at active site to reduce the hydrolysis products is possible by using the phage display approach.
format	Conference or Workshop Item
author	Abdul Kudos, Muhammad Badri Gissel, Sabrina Bordusa, Frank
author_facet	Abdul Kudos, Muhammad Badri Gissel, Sabrina Bordusa, Frank
author_sort	Abdul Kudos, Muhammad Badri
title	Selection of potentially optimized trypsin variant with a more hydrophobic cluster active site via a phage display approach
title_short	Selection of potentially optimized trypsin variant with a more hydrophobic cluster active site via a phage display approach
title_full	Selection of potentially optimized trypsin variant with a more hydrophobic cluster active site via a phage display approach
title_fullStr	Selection of potentially optimized trypsin variant with a more hydrophobic cluster active site via a phage display approach
title_full_unstemmed	Selection of potentially optimized trypsin variant with a more hydrophobic cluster active site via a phage display approach
title_sort	selection of potentially optimized trypsin variant with a more hydrophobic cluster active site via a phage display approach
publishDate	2015
url	http://irep.iium.edu.my/48397/ http://irep.iium.edu.my/48397/1/Badri%20KOP.pdf http://irep.iium.edu.my/48397/2/Poster_badri_ACB2015.pdf
first_indexed	2023-09-18T21:08:40Z
last_indexed	2023-09-18T21:08:40Z
_version_	1777411104753844224

Selection of potentially optimized trypsin variant with a more hydrophobic cluster active site via a phage display approach

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