River Stone Biotech LLC

River Stone Biotech Aps

Jens Houghton-larsen

Philipp Friedrich Berninger

Franziska Grassinger

Jon Richard Heal

Joseph Michael Sheridan

Markus Schwab

Anaelle Hatsch

Esben Halkjaer Hansen

Laura Occhipinti

The present invention relates to a recombinant microbial host cell comprising an operative biosynthetic metabolic pathway capable of producing one or more compounds selected from the group consisting of L-dopa, dopamine, (S)-Norcoclaurine and derivatives thereof; said pathway comprising a heterologous L-tyrosine hydroxylase (TyrH) converting L-Tyrosine into L-dopa capable of increasing the cell production of the Compound compared to a reference L-tyrosine hydroxylase having the sequence set forth in SEQ ID NO: 58.

Recombinant host cells with improved production of L-DOPA, dopamine, S-noroclaurine or derivatives thereof

The invention relates to norcoclaurine synthases and substrate binding sites having one or more site-specific mutation which increase the activity, when compared to the wild type synthase, of the condensation of 4-HPAA and dopamine to (S)-norcoclaurine and/or 3,4-DhPAA and dopamine to (S)-norlaudanosoline. The inventors both identified specific mutations corresponding to at position 73, 75, 77, 82, 99, 114, 141, 142, 147, 152, 174 and/or 178 in the count according to SEQ ID No: 1, and sites corresponding to the binding domains defined in SEQ ID NO: 4 and 5, where the mutated increase of the activity may be positioned within these norcoclaurine synthases. These domains are conserved regions.

Norcoclaurine synthases with increased activity

Growing community of inventors

Copenhagen, Denmark

Jens Houghton-larsen