In vitro synthetic enzymatic biosystems comprised of more than five enzymes in one pot is an emerging disruptive biomanufacturing platform that can implement complicated biological reactions, surpassing the whole cells’ constraints in terms of product yield, volumetric productivity, product titer, tolerance of toxic compounds, and so on [1]. In addition to the production of hydrogen or electricity from sugars via these in vitro synthetic pathways, we apply this platform to produce a high-value nutraceutical -- inositol from less costly substrates – starch and cellulose.<br/><br/>Inositol is a member of vitamin family B and widely used in the pharmaceutical, food, feed, and cosmetic industries. Also, it is a precursor of glucaric acid after two-step enzymatic biocatalysis. Now it is produced mainly via acid hydrolysis of phytic acid. Its high price and limited supplies prevent its wide applications, especially as a feed additive and a precursor of glucaric acid, one of top DOE renewable chemicals. Here we design ATP-free non-natural enzymatic pathways (containing more than five enzymes) to produce high-yield inositol from starch, cellulose, and glucose. High-title of inositol (e.g. 80 g/L) has been produced after 72-hour reaction. It is expected that inositol production costs will be decreased by 5-10 fold to $1/kg of inositol by using thermostable enzymes. The success of industrial inositol production would establish in vitro synthetic enzymatic biosystems as one key platform of biomanufacturing 4.0.<br/>

Biosynthesis of high-value inositol via in vitro synthetic enzymatic pathways

251st National Meeting (2016)

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