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Biochemical Characterization of β-Amino Acid Incorporation in FluvirucinB2Biosynthesis

  • Author(s): Barajas, JF
  • Zargar, A
  • Pang, B
  • Benites, VT
  • Gin, J
  • Baidoo, EEK
  • Petzold, CJ
  • Hillson, NJ
  • Keasling, JD
  • et al.
Abstract

© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. 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 fluvirucinB2polyketide 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 fluvirucinB2biosynthesis. 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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