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Open Access Publications from the University of California

Biochemical Characterization of β-Amino Acid Incorporation in Fluvirucin B2 Biosynthesis.

  • Author(s): Barajas, Jesus F;
  • Zargar, Amin;
  • Pang, Bo;
  • Benites, Veronica T;
  • Gin, Jennifer;
  • Baidoo, Edward EK;
  • Petzold, Christopher J;
  • Hillson, Nathan J;
  • Keasling, Jay D
  • et al.

Naturally occurring lactams, such as the polyketide-derived macrolactams, provide a diverse class of natural products that could enhance existing chemically produced lactams. Although β-amino acid loading in the fluvirucin B2 polyketide pathway was proposed by a previously identified putative biosynthetic gene cluster, biochemical characterization of the complete loading enzymes has not been described. Here we elucidate the complete biosynthetic pathway of the β-amino acid loading pathway in fluvirucin B2 biosynthesis. We demonstrate the promiscuity of the loading pathway to utilize a range of amino acids and further illustrate the ability to introduce non-native acyl transferases to selectively transfer β-amino acids onto a polyketide synthase (PKS) loading platform. The results presented here provide a detailed biochemical description of β-amino acid selection and will further aid in future efforts to develop engineered lactam-producing PKS platforms.

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