Mutant 4-hydroxyphenylacetate 3-hydroxylases and uses thereof

US10988786B2 · kind B2 · utility

Assignees

• SYMRISE AG · DE
• Leibniz-Institut fuer Pflanzenbiochemie (IPB) · DE

Inventors

• Ludger A. Wessjohann · Bodenwerder, DE
• Susann Herrmann · Bodenwerder, DE
• Martin Dippe · Bodenwerder, DE
• Torsten GEISSLER · Einbeck, DE
• Katrin Geissler · Einbeck, DE
• Jakob Ley · Holzminden, DE

Key dates

• Filing date: Sep 30, 2016
• Grant date: Apr 27, 2021
• Expiry date: Apr 24, 2037

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Y201/01
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention relates to genetically modified enzymes obtained by rational design of the active site binding pocket of the prototypic enzyme 4-hydroxyphenylacetate 3-hydroxylase (4HPA3H) for hydroxylating a 4-hydroxyphenyl compound to yield a 3,4-dihydroxyphenyl compound and to biotechnological methods including in vivo and in vitro methods using said enzymes or catalytically active fragments thereof. Further provided is a method either using a suitable oxidase or hydroxylase further enabling the subsequent site specific methylation of the 3,4-dihydroxyphenyl compound in a coupled enzymatic reaction by providing a suitable O-methyltransferase. Finally, compositions obtainable by the aforementioned methods are disclosed.