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bioRxiv · 10.1101/2022.12.16.520786

Reductive Enzyme Cascades for Valorization of PET Deconstruction Products

Gopal, M. R.·Dickey, R. M.·Butler, N. D.·Talley, M. R.·Mohapatra, A.·Watson, M. P.·Chen, W.·Kunjapur, A. M.
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Abstract

To better incentivize the collection of plastic wastes, new chemical transformations must be developed that add value to plastic deconstruction products. Polyethylene terephthalate (PET) is a common plastic whose deconstruction through chemical or biological means has received much attention. However, a limited number of alternative products have been formed from PET deconstruction, and only a small share could serve as building blocks for alternative materials or therapeutics. Here, we demonstrate the production of useful mono-amine and diamine building blocks from known PET deconstruction products. We achieve this by designing one-pot biocatalytic transformations that are informed by the substrate specificity of an {omega}-transaminase and diverse carboxylic acid reductases (CAR) towards PET deconstruction products. We first establish that an {omega}-transaminase from Chromobacterium violaceum (cvTA) can efficiently catalyze amine transfer to potential PET-derived aldehydes to form the mono-amine para-(aminomethyl)benzoic acid (pAMBA) or the diamine para-xylylenediamine (pXYL). We then identified CAR orthologs that could perform the bifunctional reduction of TPA to terephthalaldehyde (TPAL) or the reduction of mono-(2-hydroxyethyl) terephthalic acid (MHET) to its corresponding aldehyde. After characterizing 17 CARs in vitro, we show that the CAR from Segniliparus rotundus (srCAR) had the highest observed activity on TPA. Given these newly elucidated substrate specificity results, we designed modular enzyme cascades based on coupling srCAR and cvTA in one-pot with enzymatic co-factor regeneration. When we supply TPA, we achieve a 69 {+/-} 1% yield of pXYL, which is useful as a building block for materials. When we instead supply MHET and subsequently perform base-catalyzed ester hydrolysis, we achieve 70 {+/-} 8% yield of pAMBA, which is useful for therapeutic applications and as a pharmaceutical building block. This work expands the breadth of products derived from PET deconstruction and lays the groundwork for eventual valorization of waste PET to higher-value chemicals and materials. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=117 SRC="FIGDIR/small/520786v1_ufig1.gif" ALT="Figure 1"> View larger version (26K): org.highwire.dtl.DTLVardef@1bb7386org.highwire.dtl.DTLVardef@19d8c55org.highwire.dtl.DTLVardef@68569eorg.highwire.dtl.DTLVardef@ed398_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Gopal, M. R., Dickey, R. M., Butler, N. D., Talley, M. R., Mohapatra, A., Watson, M. P., Chen, W., Kunjapur, A. M.. 2022-12-18. Reductive Enzyme Cascades for Valorization of PET Deconstruction Products. https://doi.org/10.1101/2022.12.16.520786

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