The kinetic growth study of escherichia coli in production recombinant PLA2 / Zainab Abd Rahman
Recombinant E. coli is seen here as an alternative toward production of phospholipase A2 enzymes. The recombinant PLA2 of pBADTOPO-pla2 5 and pBADTOPO-pla2 8 were successfully cloned and sequence in previous study. The clones were recognized using SDS-PAGE and the result found that the recombinant P...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2013
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| Subjects: | |
| Online Access: | https://ir.uitm.edu.my/id/eprint/109583/1/109583.PDF https://ir.uitm.edu.my/id/eprint/109583/ |
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| Summary: | Recombinant E. coli is seen here as an alternative toward production of phospholipase A2 enzymes. The recombinant PLA2 of pBADTOPO-pla2 5 and pBADTOPO-pla2 8 were successfully cloned and sequence in previous study. The clones were recognized using SDS-PAGE and the result found that the recombinant PLA2 was successfully expressed as a soluble protein by arabinose induction. Study on kinetics growth of recombinant PLA2 creates more understanding on their biological activity in which can be used for future optimization in fermentation process. The kinetics growth of recombinant PLA2 was study in shake flask culture and their growth pattern was observed through cells viability, Bradford protein assays and dry cell weight. The culture was grown using suitable medium with suitable growth condition. The catalytic activity of recombinant pBADTOPO-pla2 5 and pBADTOPO-pla2 8 was then tested through sPLA2 assay and found to display enzymatic activity. Results from kinetics study reveal that the recombinant proteins shown normal growth pattern with high protein production at log phase. |
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