Construction of a pathway to C50-ε-carotene

PLoS One. 2019 May 14;14(5):e0216729. doi: 10.1371/journal.pone.0216729. eCollection 2019.

Abstract

Substrate tolerance of bacterial cyclases has been demonstrated in various contexts, but little is known about that of plant cyclases. Here, we tested two plant ε-cyclases to convert C50-lycopene, which we previously established by rounds of directed evolution. Unlike bacterial β-cyclases, two-end cyclase from lettuce exhibited complete specificity against this molecule, indicating that this enzyme has some mechanism that exerts size-specificity. Arabidopsis one-end cyclase At-y2 showed detectable activity to C50-lycopene. Interestingly, we found that it functions as a two-end cyclase in a C50 context. Based on this observation, a possible model for substrate discrimination of this enzyme is proposed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / enzymology
  • Arabidopsis / genetics
  • Carotenoids / chemistry*
  • Carotenoids / metabolism*
  • Lyases / genetics*
  • Lyases / metabolism*
  • Metabolic Engineering*
  • Substrate Specificity

Substances

  • Carotenoids
  • Lyases

Grants and funding

This work was supported by JSPS KAKENHI Grants 15H04189, 15K14228, and 16K21725, the Hamaguchi Foundation for the Advancement of Biochemistry, the Futaba Electronics Memorial Foundation, and the Shorai Foundation for Science and Technology. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.