Improved selectivity of an engineered multi-product terpene synthase

Ryan Lauchli*, Julia Pitzer, Rebekah Z. Kitto, Karolina Z. Kalbarczyk, Kersten S. Rabe*

*Korrespondierende/r Autor/-in für diese Arbeit

Publikation: Beitrag in einer FachzeitschriftArtikelBegutachtung

Abstract

Mutation of the sesquiterpene synthase Cop2 was conducted with a high-throughput screen for the cyclization activity using a non-natural substrate. A mutant of Cop2 was identified that contained three amino acid substitutions. This mutant, 17H2, converted the natural substrate FPP into germacrene D-4-ol with 77% selectivity. This selectivity is in contrast to that of the parent enzyme in which germacrene D-4-ol is produced as 29% and α-cadinol is produced as 46% of the product mixture. The mutations
were shown to each contribute to this selectivity, and a homology model suggested that the mutations lie near to the active site though would be unlikely to be targeted for mutation by rational methods. Kinetic comparisons show that 17H2 maintains a kcat/KM of 0.62 mM−1 s−1 , which is nearly identical to that of the
parent Cop2, which had a kcat /KM of 0.58 mM−1 s−1
Originalspracheenglisch
Seiten (von - bis)4013-4020
FachzeitschriftOrganic & Biomolecular Chemistry
Jahrgang12
DOIs
PublikationsstatusVeröffentlicht - 2014
Extern publiziertJa

Fields of Expertise

  • Human- & Biotechnology

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