BIROn - Birkbeck Institutional Research Online

    The acceptance and kinetic resolution of alpha-Methyl Substituted Aldehydes by Norcoclaurine Synthase

    Roddan, Rebecca and Gygli, G. and Sula, Altin and Méndez-Sánchez, D. and Pleiss, J. and Ward, J.M. and Keep, Nicholas H. and Hailes, H.C. (2019) The acceptance and kinetic resolution of alpha-Methyl Substituted Aldehydes by Norcoclaurine Synthase. ACS Catalysis , ISSN 2155-5435. (In Press)

    [img] Text
    Manuscript file accepted.pdf - Author's Accepted Manuscript
    Restricted to Repository staff only until 11 September 2020.

    Download (5MB) | Request a copy
    [img] Text
    RR_Methods_190909 accepted.pdf - Supplemental Material
    Restricted to Repository staff only until 11 September 2020.

    Download (7MB) | Request a copy

    Abstract

    Norcoclaurine synthase (NCS) catalyzes a stereoselective Pictet-Spengler reaction to give the key intermediate, (S)-norcoclaurine in benzylisoquinoline alkaloid (BIA) biosynthesis. This family of alkaloids contains many bioactive molecules including morphine and berberine. Recently, NCS has been demonstrated to accept a variety of aldehydes and some ketones as substrates, leading to a range of chiral tetrahydroisoquinoline (THIQ) products. Here, we report the unusual acceptance of a-substituted aldehydes, in particular a-methyl substituted aldehydes, by wild-type Thalictrum flavum NCS (Δ33TfNCS) to give THIQ products. Moreover, the kinetic resolution of several a-substituted aldehydes to give THIQs with two defined chiral centers in a single step with high conversions was achieved. Several dopamine analogues were also accepted as substrates and reactions were amenable to scale-up. Active site mutants of TfNCS were then used which demonstrated the potential to enhance the stereoselectivities in the reaction and improve yields. Rationale for the acceptance of these substrates and improved activity with different mutants has been gained from a co-crystallized structure of Δ33TfNCS with a non-productive mimic of a reaction intermediate bound in the active site. Finally, molecular dynamics simulations were performed to study the binding of dopamine and an a-substituted aldehyde and provided further insight into the reaction with these substrates.

    Metadata

    Item Type: Article
    School: Birkbeck Schools and Departments > School of Science > Biological Sciences
    Depositing User: Nick Keep
    Date Deposited: 16 Sep 2019 08:14
    Last Modified: 16 Feb 2020 18:54
    URI: http://eprints.bbk.ac.uk/id/eprint/28917

    Statistics

    Downloads
    Activity Overview
    3Downloads
    51Hits

    Additional statistics are available via IRStats2.

    Archive Staff Only (login required)

    Edit/View Item Edit/View Item