Malaria, the most severe parasitic infection in the world, caused more than one million deaths annually (Liu C et al., 2006; Bloland, P.B., 2001; World Health Organization, 2006). Artemisinin, extracted from a Chinese medicinal plant Artemisia annua L. is highly effective against multidrug-resistant Plasmodium spp. (Luo & Shen, 1987). Figure 1 shows the chemical structure of artemisinin (Liu C, 2006).
Fig 1. Chemical structure of artemisinin (Liu C, 2006)
1.1 Biosynthesis of Artemisinin
One the one hand, it is really difficult to predict the eventual supply because of the weather and overall harvest (Mutabingwa, T.K., 2005). On the other hand, it is unrealistic to use chemical methods to synthesis artemisinin because of the complexity and low yield (Liu C, 2006). The total chemical synthesis of artemisinin can be replaced by the combination of the both chemical synthesis and biosynthetic pathway (Liu et al. 2005). The chemical synthesis is to generate artemisinin from dihydroartemisinic acid (Liu C, 2006).
1.2 Engineering Cell
E. coli is one of the earliest engineered cells. There are great many advantages of E. coli used as engineered cells, such as clear genetic background, fast-growing, short growth cycle, suitable for expression of various gene products and low cost. All of these advantages become the reasons for E. coli to become the preferred host strain in artemisinin synthetic biology research (Kong, J.Q., 2013).
1.3 Feedstock for the process
Westfall (2012) proved that the production of amorpha -4, 11-diene significantly increased while mix ethanol and glucose as the carbon source for the culture medium. In addition, the restrictions on the glucose and phosphate, can effectively increase the biomass e...
... middle of paper ...
...ynthetase, isocitrate dehydrogenase and oxoglutarate dehydrogenase complex is the rate-limiting enzyme in this process and they can control the pyruvate generation. We can also decrease the metabolic side reaction through these two aspects are the necessary conditions to obtain high yields of pyruvate.
Figure 8. TCA/Citric acid cycle (Taguchi, T., 2007)
The third step is the artemisinin biosynthesis. During this period, in order to maintain the stability of the plasmids and obtain more purpose product, we need to culture E .coli to the purpose number and then use Isopropyl β-D-Thiogalactoside (IPTG). IPTG is a structural mimic of lactose, which can bind to the lac repressor and induces a similar conformational change. Then, lac repressor can no longer bind the operator and RNA polymerase of E .coli begins transcribing (Gilbert, W. & Müller-Hill, B., 1966).
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