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1000
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Abstract/Summary
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<jats:p>Cell-free systems have been used to synthesize chemicals by reconstitution of <jats:italic>in vitro</jats:italic> expressed enzymes. However, coexpression of multiple enzymes to reconstitute long enzymatic pathways is often problematic due to resource limitation/competition (e.g., energy) in the one-pot cell-free reactions. To address this limitation, here we aim to design a modular, cell-free platform to construct long biosynthetic pathways for tunable synthesis of value-added aromatic compounds, using (<jats:italic>S</jats:italic>)-1-phenyl-1,2-ethanediol ((<jats:italic>S</jats:italic>)-PED) and 2-phenylethanol (2-PE) as models. Initially, all enzymes involved in the biosynthetic pathways were individually expressed by an <jats:italic>E. coli</jats:italic>-based cell-free protein synthesis (CFPS) system and their catalytic activities were confirmed. Then, three sets of enzymes were coexpressed in three cell-free modules and each with the ability to complete a partial pathway. Finally, the full biosynthetic pathways were reconstituted by mixing two related modules to synthesize (<jats:italic>S</jats:italic>)-PED and 2-PE, respectively. After optimization, the final conversion rates for (<jats:italic>S</jats:italic>)-PED and 2-PE reached 100 and 82.5%, respectively, based on the starting substrate of <jats:sc>l</jats:sc>-phenylalanine. We anticipate that the modular cell-free approach will make a possible efficient and high-yielding biosynthesis of value-added chemicals.</jats:p>
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1000
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Hinweis
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DeepGreen-ID: b5fbae95dbe840239a407f6041a69394 ; metadata provieded by: DeepGreen (https://www.oa-deepgreen.de/api/v1/), LIVIVO search scope life sciences (http://z3950.zbmed.de:6210/livivo), Crossref Unified Resource API (https://api.crossref.org/swagger-ui/index.html), to.science.api (https://frl.publisso.de/), ZDB JSON-API (beta) (https://zeitschriftendatenbank.de/api/), lobid - Dateninfrastruktur für Bibliotheken (https://lobid.org/resources/search)
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