title: Production of mesaconate in Escherichia coli by engineered glutamate mutase pathway.
abstract: Mesaconate is an intermediate in the glutamate degradation pathway of microorganisms such as Clostridium tetanomorphum. However, metabolic engineering to produce mesaconate has not been reported previously. In this work, two enzymes involved in mesaconate production, glutamate mutase and 3-methylaspartate ammonia lyase from C. tetanomorphum, were recombinantly expressed in Escherichia coli. To improve mesaconate production, reactivatase of glutamate mutase was discovered and adenosylcobalamin availability was increased. In addition, glutamate mutase was engineered to improve the in vivo activity. These efforts led to efficient mesaconate production at a titer of 7.81 g/L in shake flask with glutamate feeding. Then a full biosynthetic pathway was constructed to produce mesaconate at a titer of 6.96 g/L directly from glucose. In summary, we have engineered an efficient system in E. coli for the biosynthesis of mesaconate.
authors.affiliation: Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Jingyu
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, MN 55455, USA. Electronic address: kzhang@umn.edu.
authors.forename: Kechun
authors.initials: K
authors.lastname: Zhang
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D048012:Clostridium tetanomorphum / Q000201:enzymology / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005650:Fumarates / Q000378:metabolism*; D019751:Intramolecular Transferases / Q000096:biosynthesis* / Q000235:genetics; D008298:Maleates / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D005650:Fumarates; D008298:Maleates; C073341:citraconic acid; D019751:Intramolecular Transferases; C021570:methylaspartate mutase
keywords: 3-Methylaspartate ammonia lyase; Adenosylcobalamin; Escherichia coli; Glutamate mutase; Mesaconate