CLONING AND HETEROLOGOUS EXPRESSION OF THE MEVALONATE PATHWAY FOR ARTEMISININ PRECURSOR PRODUCTION IN ESCHERICHIA COLI
The World Health Organization (WHO) recommends the artemisinin as the main treatment for malaria infection by Plasmodium falciparum. This compound is used together with one or more drugs in the artemisinin combination therapy (ACTs). Semi-synthetic production is a potential alternative to produce...
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| Format: | Theses |
| Language: | Indonesia |
| Online Access: | https://digilib.itb.ac.id/gdl/view/71133 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | The World Health Organization (WHO) recommends the artemisinin as the main
treatment for malaria infection by Plasmodium falciparum. This compound is used
together with one or more drugs in the artemisinin combination therapy (ACTs).
Semi-synthetic production is a potential alternative to produce artemisinin. In a
previous study, an attempt was made to increase amorphadiene production through
heterologous expression of the mevalonate pathway in B. subtilis. However, the
upper mevalonate pathway still does not work optimally. Thus, optimization is
needed to increase the expression of the upper pathway as a supply of mevalonate
intermediate products. In this study three key genes composing the mevalonate
pathway (atoB, mvaS, mvaA) were cloned into the pDR111 plasmid using the
circular polymerase extension cloning (CPEC) method. Based on the results of
restriction analysis and sequencing, cloning was successfully carried out. The
results of the SDS-PAGE expression analysis showed that there was protein
expression at the size of the target proteins atoB (40.35 kDa), mvaS (43.22 kDa)
and mvaA (46.18 kDa). Metabolite analysis by LC-MS/MS showed the presence of
mevalonate metabolites formed in cultures containing recombinant plasmids.
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