Production of glucaric acid from a synthetic pathway in recombinant Escherichia coli.
A synthetic pathway has been constructed for the production of glucuronic and glucaric acids from glucose in Escherichia coli. Coexpression of the genes encoding myo-inositol-1-phosphate synthase (Ino1) from Saccharomyces cerevisiae and myo-inositol oxygenase (MIOX) from mice led to production of glucuronic acid through the intermediate myo-inositol. Glucuronic acid concentrations up to 0.3 g/liter were measured in the culture broth. The activity of MIOX was rate limiting, resulting in the accumulation of both myo-inositol and glucuronic acid as final products, in approximately equal concentrations. Inclusion of a third enzyme, uronate dehydrogenase (Udh) from Pseudomonas syringae, facilitated the conversion of glucuronic acid to glucaric acid. The activity of this recombinant enzyme was more than 2 orders of magnitude higher than that of Ino1 and MIOX and increased overall flux through the pathway such that glucaric acid concentrations in excess of 1 g/liter were observed. This represents a novel microbial system for the biological production of glucaric acid, a "top value-added chemical" from biomass.
Moon
Tae Seok
TS
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, USA.
Yoon
Sang-Hwal
SH
Lanza
Amanda M
AM
Roy-Mayhew
Joseph D
JD
Prather
Kristala L Jones
KL
D000445:Aldehyde Oxidoreductases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005937:Glucaric Acid / Q000378:metabolism*; D007294:Inositol / Q000378:metabolism; D050562:Inositol Oxygenase / Q000235:genetics / Q000378:metabolism; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D051379:Mice; D007296:Myo-Inositol-1-Phosphate Synthase / Q000235:genetics / Q000378:metabolism; D044224:Pseudomonas syringae / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics
D011994:Recombinant Proteins; D007294:Inositol; C023280:uronate dehydrogenase; D050562:Inositol Oxygenase; D000445:Aldehyde Oxidoreductases; D007296:Myo-Inositol-1-Phosphate Synthase; D005937:Glucaric Acid