Carboxyl Methyltransferase Catalysed Formation of Mono‐ and Dimethyl Esters under Aqueous Conditions: Application in Cascade Biocatalysis

Ward, Lucy C; McCue, Hannah V.; Rigden, Daniel J.; Kershaw, Neil M.; Ashbrook, Chloe; Hatton, Harry; Goulding, Ellie; Johnson, James R.; Carnell, Andrew J.

doi:10.1002/anie.202117324

Cited by 10 publications

(24 citation statements)

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“… A recent paper also shows that mmTPA can be produced from TPA with high selectivity using a carboxyl methyltransferase from A. fumigatus . When we tested CAR activity on mmTPA, the general trend across the enzyme family was that CARs exhibited high activity on mmTPA.…”

Section: Results and Discussionmentioning

confidence: 83%

“…58 A recent paper also shows that mmTPA can be produced from TPA with high selectivity using a carboxyl methyltransferase from A. fumigatus. 59 When we tested CAR activity on mmTPA, the general trend across the enzyme family was that CARs exhibited high activity on mmTPA. While srCAR only exhibited a 1.7-fold increase in initial activity on mmTPA when compared to its activity on TPA, niCAR (from N. iowensis) exhibited a noteworthy 9.9-fold increase in activity compared to its activity on TPA.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

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Reductive Enzyme Cascades for Valorization of Polyethylene Terephthalate Deconstruction Products

et al. 2023

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To better incentivize the collection of plastic wastes, 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 monoamine 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 ω-transaminase and diverse carboxylic acid reductases (CAR) toward PET deconstruction products. We first establish that an ω-transaminase from Chromobacterium violaceum (cvTA) can efficiently catalyze amine transfer to potential PET-derived aldehydes to form monoamine para-(aminomethyl)benzoic acid (pAMBA) or diamine para-xylylenediamine (pXYL). We then identified CAR orthologs that could perform the bifunctional reduction of terephthalic acid (TPA) to terephthalaldehyde 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 elucidated substrate specificity results, we designed modular enzyme cascades based on coupling srCAR and cvTA in one pot with enzymatic cofactor regeneration. When we supply TPA, we achieve a 69 ± 1% yield of pXYL, which is useful as a building block for polymeric 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.

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Section: Results and Discussionmentioning

confidence: 83%

Section: ■ Results and Discussionmentioning

confidence: 99%

Reductive Enzyme Cascades for Valorization of Polyethylene Terephthalate Deconstruction Products

et al. 2023

View full text Add to dashboard Cite

show abstract

“…60 A recent paper also shows that mmTPA can be produced from TPA with high selectivity using a carboxyl methyltransferase from Aspergillus fumigatus. 61 When we tested CAR activity on mmTPA, the general trend across the enzyme family was that CARs exhibited high activity on mmTPA. While srCAR only exhibited a 1.7-fold increase in initial activity on mmTPA when compared to its activity on TPA, niCAR (from Nocardia iowensis) exhibited a noteworthy 9.9-fold increase in activity compared to its activity on TPA.…”

Section: Productsmentioning

confidence: 99%

Reductive Enzyme Cascades for Valorization of PET Deconstruction Products

Gopal

Dickey

Butler

et al. 2022

Preprint

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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 ω-transaminase and diverse carboxylic acid reductases (CAR) towards PET deconstruction products. We first establish that an ω-transaminase fromChromobacterium 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 fromSegniliparus 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.

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“…Webina thus simplifies the docking process, making it accessible even to novices. Though only recently published, several researchers have already used Webina to study compounds with analgesic [ 62 ], anti-cancer [ 63 , 64 ], anti-bacterial [ 65 ], anti-viral [ 66 , 67 , 68 ], and antioxidant [ 69 ] properties, among others [ 70 , 71 ]. The Webina app and source code are freely available online under the terms of the open-source Apache License, version 2.0 ( Table 1 ).…”

Section: Examples Of Cadd Browser Appsmentioning

confidence: 99%

Open-Source Browser-Based Tools for Structure-Based Computer-Aided Drug Discovery

Wang

Durrant

2022

Molecules

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We here outline the importance of open-source, accessible tools for computer-aided drug discovery (CADD). We begin with a discussion of drug discovery in general to provide context for a subsequent discussion of structure-based CADD applied to small-molecule ligand discovery. Next, we identify usability challenges common to many open-source CADD tools. To address these challenges, we propose a browser-based approach to CADD tool deployment in which CADD calculations run in modern web browsers on users’ local computers. The browser app approach eliminates the need for user-initiated download and installation, ensures broad operating system compatibility, enables easy updates, and provides a user-friendly graphical user interface. Unlike server apps—which run calculations “in the cloud” rather than on users’ local computers—browser apps do not require users to upload proprietary information to a third-party (remote) server. They also eliminate the need for the difficult-to-maintain computer infrastructure required to run user-initiated calculations remotely. We conclude by describing some CADD browser apps developed in our lab, which illustrate the utility of this approach. Aside from introducing readers to these specific tools, we are hopeful that this review highlights the need for additional browser-compatible, user-friendly CADD software.

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Carboxyl Methyltransferase Catalysed Formation of Mono‐ and Dimethyl Esters under Aqueous Conditions: Application in Cascade Biocatalysis

Cited by 10 publications

References 50 publications

Reductive Enzyme Cascades for Valorization of Polyethylene Terephthalate Deconstruction Products

Reductive Enzyme Cascades for Valorization of Polyethylene Terephthalate Deconstruction Products

Reductive Enzyme Cascades for Valorization of PET Deconstruction Products

Open-Source Browser-Based Tools for Structure-Based Computer-Aided Drug Discovery

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