ENGINEERING OF DEOXYXYLULOSE PHOSPHATE PATHWAY FOR ISOPRENOID PRODUCTION

Abstract

Metabolic engineering of the deoxyxylulose phosphate (DXP) pathway in microbes has been demonstrated to be an efficient and sustainable strategy to produce isoprenoid compounds from natural abundant carbon sources, which potentially could alleviate the shortage in supply of many isoprenoid pharmaceuticals. So far, the mechanisms underlying the successful metabolic engineering of the DXP pathway are not fully understood, leading to numerous trial and error approaches in enhancing the production of these metabolites. In this thesis, essential analytical tools were developed and applied to interrogate the biochemical mechanisms, which eventually led to the invention of many novel rational approaches for better engineering of the DXP pathway. In this thesis, three major projects were presented: 1) Development of an accurate transcription quantification method in metabolic engineered Escherichia coli; 2) Invention of a mass spectrometry based method for quantifying the DXP metabolites; 3) Investigation of protein insolubility of the DXP enzymes.