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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.
Abstract

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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