Screening of β-glucoside kinase directed evolution library to form a sulfur-phosphorus bond

Bellag, Oualid Abdelkader and Ben Belgacem, Farah Fadwa and Noor Batcha, Ibrahim Ali and Mohd Salleh, Hamzah (2016) Screening of β-glucoside kinase directed evolution library to form a sulfur-phosphorus bond. In: 4th International Conference on Biotechnology Engineering 2016 (ICBioE 2016), 25th-27th July 2016, Kuala Lumpur.

PDF - Published Version Restricted to Repository staff only Download (1MB) | Request a copy

Abstract

Glycoscience is a field that explores the structures and functions of glycans promising great advances in an array of applications. The objective of this research is to form a sulfur-phosphorus bond at the position 6’ of the sugar involved in many biological processes. A directed evolution strategy was adopted with error-prone PCR technique to create random mutagenesis of recombinant plasmid library of the β-glucoside kinase (βglk) recloned into a pTrcHisB plasmid and transformed into Escherichia coli. Following the creation of libraries, high throughput screening by 96-well microplate using fluorescent activated cell sorting (FACS) techniques were carried out. Positive clones were selected, sequenced for identification and enzyme expression and characterization were carried out.

Item Type:	Conference or Workshop Item (Invited Papers)
Additional Information:	3704/51693
Uncontrolled Keywords:	Directed Evolution; error-prone PCR; Fluorescent Activated Cell Sorting (FACS); β-glucoside kinase (βglk)
Subjects:	T Technology > TP Chemical technology > TP248.13 Biotechnology
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Engineering
Depositing User:	Prof. Dr. Ibrahim Ali Noorbatcha
Date Deposited:	19 Aug 2016 10:05
Last Modified:	19 Aug 2016 10:05
URI:	http://irep.iium.edu.my/id/eprint/51693

Actions (login required)

View Item