Screening of β-glucoside kinase directed evolution library to form a sulfur-phosphorus bond
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 strateg...
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| Main Authors: | , , , |
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| Format: | Conference or Workshop Item |
| Language: | English |
| Published: |
Kulliyah of Engineering, International Islamic University Malaysia
2016
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/51693/1/51693.pdf http://irep.iium.edu.my/51693/ http://www.iium.edu.my/icbioe/2016/index.php/78-the-4th-international-conference-on-biotechnology-engineering |
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| Summary: | 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. |
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