Identification of novel transaminases from a 12‐aminododecanoic acid‐metabolizing Pseudomonas strain

Microbial Biotechnology
2015.0

Abstract

<jats:title>Summary</jats:title><jats:p>A <jats:styled-content style="fixed-case"><jats:italic>P</jats:italic></jats:styled-content><jats:italic>seudomonas</jats:italic> species [<jats:styled-content style="fixed-case"><jats:italic>P</jats:italic></jats:styled-content><jats:italic>seudomonas</jats:italic> sp. strain amino alkanoate catabolism (<jats:styled-content style="fixed-case">AAC</jats:styled-content>)] was identified that has the capacity to use 12‐aminododecanoic acid, the constituent building block of homo‐nylon‐12, as a sole nitrogen source. Growth of <jats:styled-content style="fixed-case"><jats:italic>P</jats:italic></jats:styled-content><jats:italic>seudomonas</jats:italic> sp. strain <jats:styled-content style="fixed-case">AAC</jats:styled-content> could also be supported using a range of additional ω‐amino alkanoates. This metabolic function was shown to be most probably dependent upon one or more transaminases (<jats:styled-content style="fixed-case">TAs</jats:styled-content>). Fourteen genes encoding putative <jats:styled-content style="fixed-case">TAs</jats:styled-content> were identified from the genome of <jats:styled-content style="fixed-case"><jats:italic>P</jats:italic></jats:styled-content><jats:italic>seudomonas</jats:italic> sp. <jats:styled-content style="fixed-case">AAC</jats:styled-content>. Each of the 14 genes was cloned, 11 of which were successfully expressed in <jats:styled-content style="fixed-case"><jats:italic>E</jats:italic></jats:styled-content><jats:italic>scherichia coli</jats:italic> and tested for activity against 12‐aminododecanoic acid. In addition, physiological functions were proposed for 9 of the 14 <jats:styled-content style="fixed-case">TAs</jats:styled-content>. Of the 14 proteins, activity was demonstrated in 9, and of note, 3 <jats:styled-content style="fixed-case">TAs</jats:styled-content> were shown to be able to catalyse the transfer of the ω‐amine from 12‐aminododecanoic acid to pyruvate. Based on this study, three enzymes have been identified that are promising biocatalysts for the production of nylon and related polymers.

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