1. Biochemical Characterization of β‐Amino Acid Incorporation in Fluvirucin B 2 Biosynthesis
- Author
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Nathan J. Hillson, Jesus F. Barajas, Christopher J. Petzold, Veronica T. Benites, Jennifer W. Gin, Edward E. K. Baidoo, Amin Zargar, Bo Pang, and Jay D. Keasling
- Subjects
0301 basic medicine ,Fluvirucin B2 ,Lactams ,Biosynthesis ,Biochemistry ,Fluvirucin ,03 medical and health sciences ,chemistry.chemical_compound ,Polyketide ,Transferases ,Polyketide synthase ,Gene cluster ,polycyclic compounds ,Molecular Biology ,chemistry.chemical_classification ,biology ,Organic Chemistry ,Acid load ,Amino acid ,030104 developmental biology ,Enzyme ,chemistry ,Polyketides ,biology.protein ,Molecular Medicine - 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.
- Published
- 2018