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DNA assembly techniques for next-generation combinatorial biosynthesis of natural products

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Journal of Industrial Microbiology & Biotechnology

Abstract

Natural product scaffolds remain important leads for pharmaceutical development. However, transforming a natural product into a drug entity often requires derivatization to enhance the compound’s therapeutic properties. A powerful method by which to perform this derivatization is combinatorial biosynthesis, the manipulation of the genes in the corresponding pathway to divert synthesis towards novel derivatives. While these manipulations have traditionally been carried out via restriction digestion/ligation-based cloning, the shortcomings of such techniques limit their throughput and thus the scope of corresponding combinatorial biosynthesis experiments. In the burgeoning field of synthetic biology, the demand for facile DNA assembly techniques has promoted the development of a host of novel DNA assembly strategies. Here we describe the advantages of these recently developed tools for rapid, efficient synthesis of large DNA constructs. We also discuss their potential to facilitate the simultaneous assembly of complete libraries of natural product biosynthetic pathways, ushering in the next generation of combinatorial biosynthesis.

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Acknowledgments

We thank the National Institutes of Health (GM077596), the National Academies Keck Futures Initiative on Synthetic Biology, and the Energy Biosciences Institute for financial support in our development and application of DNA assembly technologies. REC acknowledges the Drickamer Fellowship support from the Department of Chemical and Biomolecular Engineering at the University of Illinois.

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Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA

    Ryan E. Cobb, Jonathan C. Ning & Huimin Zhao

  2. Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA

    Ryan E. Cobb & Huimin Zhao

  3. Departments of Chemistry, Biochemistry, and Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA

    Huimin Zhao

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  1. Ryan E. Cobb
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  2. Jonathan C. Ning
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Correspondence to Huimin Zhao.

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Cobb, R.E., Ning, J.C. & Zhao, H. DNA assembly techniques for next-generation combinatorial biosynthesis of natural products. J Ind Microbiol Biotechnol 41, 469–477 (2014). https://doi.org/10.1007/s10295-013-1358-3

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  • DOI: https://doi.org/10.1007/s10295-013-1358-3

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