Construction of an Efficient Escherichia Coli Whole-Cell Biocatalyst for D-mannitol Production

J Biosci Bioeng. 2014 Dec;118(6):628-31. doi: 10.1016/j.jbiosc.2014.05.004. Epub 2014 Jun 5.

Abstract

Mannitol is a six carbon sugar alcohol that finds applications in the pharmaceutical and food industries. A novel Escherichia coli strain capable of converting D-glucose to D-mannitol has been constructed, wherein native mannitol-1-phosphate dehydrogenase (MtlD) and codon-optimized Eimeria tenella mannitol-1-phosphatase (M1Pase) have been overexpressed. Codon-optimized Pseudomonas stutzeri phosphite dehydrogenase (PtxD) was overexpressed for cofactor (NADH) regeneration with the concomitant oxidation of phosphite to phosphate. Whole-cell biotransformation using resting cells in a medium containing D-glucose and equimolar sodium phosphite resulted in d-mannitol yield of 87 mol%. Thus, production of an industrially relevant biochemical without using complex media components and elaborate process control mechanisms has been demonstrated.

Keywords: Biotransformation; Cofactor regeneration; Resting cells; Whole-cell biocatalyst; d-Mannitol production.

MeSH terms

  • Biocatalysis*
  • Biosynthetic Pathways / genetics
  • Biotransformation
  • Codon / genetics
  • Eimeria tenella / enzymology
  • Escherichia coli / cytology*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Fructosephosphates / metabolism
  • Genetic Engineering
  • Glucose / metabolism
  • Mannitol / chemistry
  • Mannitol / metabolism*
  • NAD / metabolism
  • NADH, NADPH Oxidoreductases / genetics
  • NADH, NADPH Oxidoreductases / metabolism
  • Oxidation-Reduction
  • Phosphites / metabolism
  • Phosphoric Monoester Hydrolases / genetics
  • Phosphoric Monoester Hydrolases / metabolism
  • Pseudomonas stutzeri / enzymology
  • Sugar Alcohol Dehydrogenases / metabolism

Substances

  • Codon
  • Fructosephosphates
  • Phosphites
  • NAD
  • Mannitol
  • fructose-6-phosphate
  • Sugar Alcohol Dehydrogenases
  • mannitol-1-phosphate dehydrogenase
  • NADH, NADPH Oxidoreductases
  • NAD phosphite oxidoreductase
  • Phosphoric Monoester Hydrolases
  • mannitol-1-phosphatase
  • Glucose