Abstract
One of the most abundant phenolic compounds traced in olive tissues is hydroxytyrosol (HT), a molecule that has been attributed with a pile of beneficial effects, well documented by many epidemiological studies and thus adding value to products containing it. Strong antioxidant capacity and protection from cancer are only some of its exceptional features making it ideal as a potential supplement or preservative to be employed in the nutraceutical, agrochemical, cosmeceutical, and food industry. The HT biosynthetic pathway in plants (e.g. olive fruit tissues) is not well apprehended yet. In this contribution we employed a metabolic engineering strategy by constructing a dual pathway introduced in Escherichia coli and proofing its significant functionality leading it to produce HT. Our primary target was to investigate whether such a metabolic engineering approach could benefit the metabolic flow of tyrosine introduced to the conceived dual pathway, leading to the maximalization of the HT productivity. Various gene combinations derived from plants or bacteria were used to form a newly inspired, artificial biosynthetic dual pathway managing to redirect the carbon flow towards the production of HT directly from glucose. Various biosynthetic bottlenecks faced due to feaB gene function, resolved through the overexpression of a functional aldehyde reductase. Currently, we have achieved equimolar concentration of HT to tyrosine as precursor when overproduced straight from glucose, reaching the level of 1.76 mM (270.8 mg/L) analyzed by LC-HRMS. This work realizes the existing bottlenecks of the metabolic engineering process that was dependent on the utilized host strain, growth medium as well as to other factors studied in this work.
Highlights
Hydroxytyrosol (3,4-Dihydroxyphenylethanol; HT) is a mono-phenolic compound traced in olive fruits [1] and tissues [2], in extracted olive oil [3], or even in olive mills waste waters [4]
According to Juminaga et al [29], six of these genes were necessary for the biosynthesis of shikimic acid and five of them required for the conversion of shikimic acid into tyrosine
We chose a rational approach for its production [16] where the primary metabolism was engineered in order to boost tyrosine production that is subsequently utilized for the biosynthesis of HT through the engineered secondary metabolic machinery
Summary
Hydroxytyrosol (3,4-Dihydroxyphenylethanol; HT) is a mono-phenolic compound traced in olive fruits [1] and tissues [2], in extracted olive oil [3], or even in olive mills waste waters [4]. It shows a broad spectrum of biological properties due to its strong antioxidant and radical-. Metabolic engineering of Escherichia coli to produce hydroxytyrosol design, data collection and analysis, decision to publish, or preparation of the manuscript
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