Evolutionary analysis of the functional plasticity of Staphylococcus aureus C30 carotenoid synthase

Maiko Furubayashi, Kyoichi Saito, Daisuke Umeno

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

Most natural carotenoids have 40-carbon (C40) backbones, while some bacteria produce carotenoids with C30 backbones. Carotenoid backbone synthases, the enzyme that catalyze the first committed step in carotenoid biosynthesis, are known to be highly specific. Previously, using C30 backbone synthase (diapophytoene synthase, CrtM) from Staphylococcus aureus, we reported two size-shifting mutations, F26A and W38A, which confer C40 synthase activity at the cost of the original C30 synthase activity. In this study, we performed a directed evolution of the C40-specialist variant CrtMF26A in search of mutations that restore the original C30 synthase function. Examination of the resultant mutants, together with the site-directed mutagenesis study identified three new mutations (H12A, D27A and I240F) that affect the size specificity of this enzyme. After re-defining the reading frame, we obtained CrtM variants that are highly active in C30 and C40 carotenoid synthesis.

Original languageEnglish
Pages (from-to)431-436
Number of pages6
JournalJournal of Bioscience and Bioengineering
Volume117
Issue number4
DOIs
Publication statusPublished - 2014 Apr
Externally publishedYes

Keywords

  • Carotene synthase
  • Carotenoid
  • Directed evolution
  • Screening
  • Size specificity

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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