The substrate specificity, enantioselectivity and structure of the (R)‐selective amine : pyruvate transaminase from Nectria haematococca. (7th April 2014)

• Record Type: Journal Article
• Authors: Sayer, Christopher; Martinez‐Torres, Ruben J.; Richter, Nina; Isupov, Michail N.; Hailes, Helen C.; Littlechild, Jennifer A.; Ward, John M.
• Is Part Of: FEBS journal. Volume 281:Number 9(2014)
• Page Start: 2240
• Page End: 2253
• Publication Date: 2014-04-07
• Subjects: Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pathology, Molecular -- Periodicals; 572
• DOI: 10.1111/febs.12778
• Languages: English
• ISSNs: 1742-464X

Abstract

<abstract abstract-type="main" id="febs12778-abs-0001"> <title> <x xml:space="preserve">Abstract</x> </title> <sec id="febs12778-sec-0001" sec-type="section"> <p>During the last decade the use of transaminases for the production of pharmaceutical and fine chemical intermediates has attracted a great deal of attention. Transaminases are versatile biocatalysts for the efficient production of amine intermediates and many have (<italic>S</italic>)‐enantiospecificity. Transaminases with (<italic>R</italic>)‐specificity are needed to expand the applications of these enzymes in biocatalysis. In this work we have identified a fungal putative (<italic>R</italic>)‐specific transaminase from the Eurotiomycetes <italic>Nectria haematococca</italic>, cloned a synthetic version of this gene, demonstrated (<italic>R</italic>)‐selective deamination of several substrates including (<italic>R</italic>)‐α‐methylbenzylamine, as well as production of (<italic>R</italic>)‐amines, and determined its crystal structure. The crystal structures of the holoenzyme and the complex with an inhibitor gabaculine offer the first detailed insight into the structural basis for substrate specificity and enantioselectivity of the industrially important class of (<italic>R</italic>)‐selective amine : pyruvate transaminases.</p> </sec> <sec id="febs12778-sec-0002" sec-type="section"> <title>Database</title> <p>The atomic coordinates and structure factors for the <italic>Nectria </italic>TAm in holoenzyme and gabaculine‐bound forms have been deposited in the PDB as entries 4cmd and 4cmf respectively.</p> </sec> <sec id="febs12778-sec-0003" sec-type="section"> <title>Structured digital abstract</title> <p> <ext-link ext-link-type="uri" xlink:href="http://www.uniprot.org/uniprot/C7YVL8" xlink:type="simple" xmlns:xlink="http://www.w3.org/1999/xlink">TAm</ext-link> and <ext-link ext-link-type="uri" xlink:href="http://www.uniprot.org/uniprot/C7YVL8" xlink:type="simple" xmlns:xlink="http://www.w3.org/1999/xlink">TAm</ext-link><ext-link ext-link-type="uri" xlink:href="http://www.ebi.ac.uk/ontology-lookup/?termId=MI:0407%20" xlink:type="simple" xmlns:xlink="http://www.w3.org/1999/xlink">bind</ext-link> by <ext-link ext-link-type="uri" xlink:href="http://www.ebi.ac.uk/ontology-lookup/?termId=MI:0114%20" xlink:type="simple" xmlns:xlink="http://www.w3.org/1999/xlink">x-ray crystallography</ext-link> (<ext-link ext-link-type="uri" xlink:href="http://www.ebi.ac.uk/intact/interaction/EBI-9255046" xlink:type="simple" xmlns:xlink="http://www.w3.org/1999/xlink">View interaction</ext-link>)</p> </sec> </abstract>