Cell-free production of isobutanol: A completely immobilized system.

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ID: 54326
2019
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Abstract
A completely immobilized cell-free enzyme reaction system was used to convert ketoisovaleric acid to isobutanol, a desirable biofuel, with a molar yield of 43% and a titer of 2 g/L, which are comparable to high performing in vivo systems (e.g. 41% and 5.4 g/L, respectively, for Clostridium thermocellum). The approach utilizes, for the first time, a series of previously reported enzyme mutants that either overproduce the product or are more stable when compared with their wild type. The selected enzyme variants include keto-acid decarboxylase attached to a maltose binding protein, alcohol dehydrogenase, and formate dehydrogenase. These enzymes were screened for thermal, pH, and product stability to choose optima for this system which were pH 7.4 and 35 °C. This system is designed to address well-known limitations of in vivo systems such as low product concentrations due to product feedback inhibition, instability of cells, and lack of economic product recovery.
Reference Key
wong2019cellfreebioresource Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Wong, Matthew;Zha, Jian;Sorci, Mirco;Gasparis, Christopher;Belfort, Georges;Koffas, Mattheos;
Journal Bioresource technology
Year 2019
DOI
S0960-8524(19)31334-3
URL
Keywords
biofuels immobilized enzymes mutagenesis isobutanol in vitro production

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