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Engineering Natural Product Biosynthesis pp 223–238Cite as

Understanding and Manipulating Assembly Line Biosynthesis by Heterologous Expression in Streptomyces

Part of the Methods in Molecular Biology book series (MIMB,volume 2489)

Abstract

Assembly line enzymes, including polyketide synthases and nonribosomal peptide synthetases, play central roles in the construction of complex natural products. Due to the sequential biochemistry processed in each domain, the domain architecture of the assembly line enzymes strictly correlates with the product molecule. This colinearity makes assembly line enzymes an ideal target for rational reprogramming. Although many of the past engineering attempts suffered from decreased product yield, recent advancements in the bioinformatic analysis and engineering design now provide new opportunity to work on these modular megaenzymes. This chapter describes the methods for analyzing and engineering the assembly line enzymes, including module and domain analysis needed for designing the engineering of assembly line biosynthesis, and the expression vector construction with an example of two-vector heterologous expression system in Streptomyces.

Key words

  • Assembly line engineering
  • Heterologous expression
  • Bioinformatic analysis
  • Streptomyces
  • Biosynthetic pathway

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Acknowledgments

This work was supported by National Natural Science Foundation for Young Scientists of China (82003631) and National Natural Science Foundation of China General Program (22177092) to L.Z., Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan (JSPS KAKENHI Grant Number JP16H06443, JP 20KK0173, and JP20H00490), Japan Science and Technology Agency (JST SICORP Grant No. JPMJSC1701), the New Energy and Industrial Technology Development Organization (NEDO, Grant Number JPNP20011), and AMED (Grant Number JP21ak0101164) to I.A., and JP17H04763, JP19H04641, JP21H02636, UTEC-UTokyo FSI Research Grant Program, Kato Memorial Bioscience Foundation, and The Asahi Glass Foundation, to T.A.

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Authors and Affiliations

  1. Key Laboratory of Precise Synthesis of Functional Molecules of Zhejiang Province, School of Science, Westlake University, Hangzhou, Zhejiang Province, China

    Lihan Zhang

  2. Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan

    Takayoshi Awakawa & Ikuro Abe

  3. Collaborative Research Institute for Innovative Microbiology, The University of Tokyo, Tokyo, Japan

    Takayoshi Awakawa & Ikuro Abe

Authors
  1. Lihan Zhang
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  2. Takayoshi Awakawa
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  3. Ikuro Abe
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Corresponding authors

Correspondence to Lihan Zhang or Ikuro Abe .

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Editors and Affiliations

  1. Department of Chemistry & BioDiscovery Institute, University of North Texas, Denton, USA

    Dr. Elizabeth Skellam

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Zhang, L., Awakawa, T., Abe, I. (2022). Understanding and Manipulating Assembly Line Biosynthesis by Heterologous Expression in Streptomyces. In: Skellam, E. (eds) Engineering Natural Product Biosynthesis. Methods in Molecular Biology, vol 2489. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2273-5_12

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  • DOI: https://doi.org/10.1007/978-1-0716-2273-5_12

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