Evolution of the C30 carotenoid synthase CrtM for function in a C40 pathway

Daisuke Umeno*, Alexander V. Tobias, Frances H. Arnold

*この研究の対応する著者

研究成果: Article査読

60 被引用数 (Scopus)

抄録

The C30 carotene synthase CrtM from Staphylococcus aureus and the C40 carotene synthase CrtB from Erwinia uredovora were swapped into their respective foreign C40 and C30 biosynthetic pathways (heterologously expressed in Escherichia coli) and evaluated for function. Each displayed negligible ability to synthesize the natural carotenoid product of the other. After one round of mutagenesis and screening, we isolated 116 variants of CrtM able to synthesize C40 carotenoids. In contrast, we failed to find a single variant of CrtB with detectable C30 activity. Subsequent analysis revealed that the best CrtM mutants performed comparably to CrtB in an in vivo C40 pathway. These mutants showed significant variation in performance in their original C30 pathway, indicating the emergence of enzymes with broadened substrate specificity as well as those with shifted specificity. We discovered that Phe 26 alone determines the specificity of CrtM. The plasticity of CrtM with respect to its substrate and product range highlights the potential for creating further new carotenoid backbone structures.

本文言語English
ページ(範囲)6690-6699
ページ数10
ジャーナルJournal of Bacteriology
184
23
DOI
出版ステータスPublished - 2002 12
外部発表はい

ASJC Scopus subject areas

  • 微生物学
  • 分子生物学

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