Biosynthesis of the Nylon 12 Monomer, ω-Aminododecanoic Acid with Novel CYP153A, AlkJ, and ω-TA Enzymes

Biotechnol J. 2018 Apr;13(4):e1700562. doi: 10.1002/biot.201700562. Epub 2018 Jan 10.

Abstract

Bioplastics are derived from renewable biomass sources, such as vegetable oils, cellulose, and starches. An important and high-performance member of the bioplastic family is Nylon 12. The biosynthesis of ω-amino dodecanoic acid (ω-AmDDA), the monomer of Nylon 12 from vegetable oil derivatives is considered as an alternative to petroleum-based monomer synthesis. In this study, for the production of ω-AmDDA from dodecanoic acid (DDA), the cascade of novel P450 (CYP153A), alcohol dehydrogenase (AlkJ), and ω-transaminase (ω-TA) is developed. The regioselective ω-hydroxylation of 1 mM DDA with near complete conversion (>99%) is achieved using a whole-cell biocatalyst co-expressing CYP153A, ferredoxin reductase and ferredoxin. When the consecutive biotransformation of ω-hydroxy dodecanoic acid (ω-OHDDA) is carried out using a whole-cell biocatalyst co-expressing AlkJ and ω-TA, 1.8 mM ω-OHDDA is converted into ω-AmDDA with 87% conversion in 3 h. Finally, when a one-pot reaction is carried out with 2 mM DDA using both whole-cell systems, 0.6 mM ω-AmDDA is produced after a 5 h reaction. The results demonstrated the scope of the potential cascade reaction of novel CYP153A, AlkJ, and ω-TA for the production of industrially important bioplastic monomers, amino fatty acids, from FFAs.

Keywords: CYP153A; Nylon 12; bioplastics; dodecanoic acid; ω-aminododecanoic acid.

MeSH terms

  • Alcohol Dehydrogenase / genetics
  • Alcohol Dehydrogenase / metabolism*
  • Amino Acids / biosynthesis*
  • Cloning, Molecular
  • Cytochrome P-450 Enzyme System / genetics
  • Cytochrome P-450 Enzyme System / metabolism*
  • Ferredoxins / metabolism
  • Lauric Acids / metabolism
  • Metabolic Engineering
  • Mycobacterium / enzymology
  • Mycobacterium / genetics
  • Recombinant Proteins / metabolism
  • Sulfite Reductase (Ferredoxin) / metabolism
  • Transaminases / genetics
  • Transaminases / metabolism*

Substances

  • Amino Acids
  • Ferredoxins
  • Lauric Acids
  • Recombinant Proteins
  • lauric acid
  • Cytochrome P-450 Enzyme System
  • omega-aminoenantic acid
  • Alcohol Dehydrogenase
  • Sulfite Reductase (Ferredoxin)
  • Transaminases