Use of directed enzyme evolution to create novel biosynthetic pathways for production of rare or non-natural carotenoids

Maiko Furubayashi*, Daisuke Umeno

*この研究の対応する著者

研究成果: Chapter

1 被引用数 (Scopus)

抄録

In recent years, advances in bioengineering and synthetic biology techniques have been used to create carotenoid diversity in the laboratory. In this chapter, we describe the step-by-step method to perform directed evolution of carotenoid biosynthetic enzymes. We first explain how to establish an efficient Escherichia coli colony-based screening, including a detailed description of plasmid DNA construction design as well as tips and tricks to handle and manipulate cells to produce stable colonies. As an example for the directed evolution experiment, we engineer a bacterial phytoene desaturase CrtI to obtain a C50-phytoene desaturase, which catalyzes formation of a non-natural long-chain carotenoid. The method described in this chapter can be applied to many carotenoid biosynthetic enzymes, whose numbers have been rapidly expanding with recent advances in genomics. The use of directed evolution for carotenoid enzymes will contribute not only to the discovery of novel carotenoids but also to a deeper understanding of the creation and evolution of carotenoid biosynthetic pathways in nature.

本文言語English
ホスト出版物のタイトルCarotenoids
ホスト出版物のサブタイトルCarotenoid and apocarotenoid biosynthesis metabolic engineering and synthetic biology
編集者Eleanore T. Wurtzel, Eleanore T. Wurtzel
出版社Academic Press Inc.
ページ351-382
ページ数32
ISBN(印刷版)9780323913539
DOI
出版ステータスPublished - 2022 1月

出版物シリーズ

名前Methods in Enzymology
671
ISSN(印刷版)0076-6879
ISSN(電子版)1557-7988

ASJC Scopus subject areas

  • 生化学
  • 分子生物学

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