| Autor: |
Gonzalez-Esquer, C Raul, Ferlez, Bryan, Weraduwage, Sarathi M, Kirst, Henning, Lantz, Alexandra T, Turmo, Aiko, Sharkey, Thomas D, Kerfeld, Cheryl A |
| Rok vydání: |
2021 |
| Předmět: |
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| Zdroj: |
RSC advances, vol 11, iss 48 |
| Popis: |
Terpene synthases are biotechnologically-relevant enzymes with a variety of applications. However, they are typically poor catalysts and have been difficult to engineer. Structurally, most terpene synthases share two conserved domains (α- and β-domains). Some also contain a third domain containing a second active site (γ-domain). Based on the three-domain architecture, we hypothesized that αβ terpene synthases could be engineered by insertion of a heterologous domain at the site of the γ-domain (an approach we term "Insertion-engineering terpene synthase"; Ie-TS). We demonstrate that by mimicking the domain architecture of αβγ terpene synthases, we can redesign isoprene synthase (ISPS), an αβ terpene synthase, while preserving enzymatic activity. Insertion of GFP or a SpyCatcher domain within ISPS introduced new functionality while maintaining or increasing catalytic turnover. This insertion-engineering approach establishes that the γ-domain position is accessible for incorporation of additional sequence features and enables the rational engineering of terpene synthases for biotechnology. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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