Description:
<jats:title>Abstract</jats:title><jats:sec>
<jats:title>Background</jats:title>
<jats:p>One carbon (C1) molecules such as methanol have the potential to become sustainable feedstocks for biotechnological processes, as they can be derived from CO<jats:sub>2</jats:sub> and green hydrogen, without the need for arable land. Therefore, we investigated the suitability of the methylotrophic yeast <jats:italic>Ogataea polymorpha</jats:italic> as a potential production organism for platform chemicals derived from methanol. We selected acetone, malate, and isoprene as industrially relevant products to demonstrate the production of compounds with 3, 4, or 5 carbon atoms, respectively.</jats:p>
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<jats:title>Results</jats:title>
<jats:p>We successfully engineered <jats:italic>O. polymorpha</jats:italic> for the production of all three molecules and demonstrated their production using methanol as carbon source. We showed that the metabolism of <jats:italic>O. polymorpha</jats:italic> is well suited to produce malate as a product and demonstrated that the introduction of an efficient malate transporter is essential for malate production from methanol. Through optimization of the cultivation conditions in shake flasks, which included pH regulation and constant substrate feeding, we were able to achieve a maximum titer of 13 g/L malate with a production rate of 3.3 g/L/d using methanol as carbon source. We further demonstrated the production of acetone and isoprene as additional heterologous products in <jats:italic>O. polymorpha</jats:italic>, with maximum titers of 13.6 mg/L and 4.4 mg/L, respectively.</jats:p>
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<jats:title>Conclusion</jats:title>
<jats:p>These findings highlight how <jats:italic>O. polymorpha</jats:italic> has the potential to be applied as a versatile cell factory and contribute to the limited knowledge on how methylotrophic yeasts can be used for the production of low molecular weight biochemicals from methanol. Thus, this study can serve as a point of reference for future metabolic engineering in <jats:italic>O. polymorpha</jats:italic> and process optimization efforts to boost the production of platform chemicals from renewable C1 carbon sources.</jats:p>
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