Team:SDU-Denmark/Tour30

The production system

The micro-road to rubber

Natural rubber is a chain of several isoprene molecules. The rubber tree has a prenyltransferase PrenyltransferaseHRT2 enzyme, which links molecules of IPP Isopentenyl pyrophosphate into the isoprene chain natural rubber. To start the extension of the chain, the prenyltransferase needs a DMAPP Dimethylallyl pyrophosphate molecule. Thus, in order to produce rubber, the bacteria needs the prenyltranferase, IPP, and DMAPP.

The basic idea is to convert sugar (glucose) into natural rubber through the glycolysis to yield pyruvate and GAP. Glyceraldehyde-3-phosphate These are further converted into IPP Isopentenyl pyrophosphate and DMAPP Dimethylallyl pyrophosphate through the MEP pathway. A DMAPP and several IPP molecules can then be linked together into natural rubber through the action of the prenyltransferase HRT2 from the rubber tree. IPP and DMAPP are molecules naturally present in E. coli, produced by the bacteria through the MEP pathway. Methylerythritol phosphate pathway The MEP pathway converts molecules of GAP Glyceraldehyde-3-phosphate and pyruvate - produced by the degradation of glucose (glycolysis) - into IPP and DMAPP. To increase the amount of IPP and DMAPP available for rubber production, we optimized the MEP pathway by elevating the levels of rate limiting enzymes in the pathway.

Through gene manipulation, we introduced the prenyltransferase gene from the rubber tree to the genetic code of E. coli. This enables the bacteria to express the prenyltransferase enzyme. Consequently the bacteria should be able to convert sugar into natural rubber through; the glycolysis, the MEP pathway, and the prenyltransferase activity.

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