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Biofuels Bioprod."],"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:p>The rising demand for natural products is accelerating research into sustainable methods for producing bio-based flavourings like ethyl butyrate. In this study, ethyl butyrate was successfully produced through the enzymatic esterification of butyric acid and ethanol, which were derived from the co-cultivation of <jats:italic>Clostridium tyrobutyricum<\/jats:italic> and <jats:italic>Saccharomyces cerevisiae<\/jats:italic>. Initial monoculture experiments with both strains were performed to investigate compromised fermentation conditions for co-cultivation. Based on these findings, anaerobic co-cultivation conditions were established at 37\u00a0\u00b0C and 150\u00a0rpm, with the pH controlled at 6. The effects of varying inoculation times in co-culture were examined, considering the solvent and acid tolerance of both strains. Due to the limited acid tolerance of <jats:italic>S. cerevisiae<\/jats:italic>, with significant inhibition at butyric acid concentrations above 10\u00a0g\u00a0L\u00af<jats:sup>1<\/jats:sup>, a time-delayed inoculation with <jats:italic>C. tyrobutyricum<\/jats:italic> was implemented. In batch experiments, the final concentrations of butyric acid and ethanol were 13.98\u2009\u00b1\u20093.06\u00a0g\u00a0L\u00af<jats:sup>1<\/jats:sup> and 21.43\u2009\u00b1\u20091.66\u00a0g\u00a0L\u00af<jats:sup>1<\/jats:sup>, respectively. Further enhancement of product concentrations was explored through a fed-batch cultivation strategy yielding up to 45.62\u2009\u00b1\u20093.82\u00a0g\u00a0L\u00af<jats:sup>1<\/jats:sup> of butyric acid and 18.61\u2009\u00b1\u20094.11\u00a0g\u00a0L\u00af<jats:sup>1<\/jats:sup> of ethanol. Ethyl butyrate was formed from the fermentation products by lipase-catalysed enzymatic esterification in a two-phase system through the addition of an organic phase. The ester concentration in the organic phase reached a maximum of 23.93\u2009\u00b1\u20090.68\u00a0g\u00a0L\u00af<jats:sup>1<\/jats:sup> (esterification yield 25%). This study presents a viable approach to the production of bio-based ethyl butyrate offering a sustainable alternative to traditional chemical synthesis methods.<\/jats:p>\n          <jats:p>\n            <jats:bold>Graphical Abstract<\/jats:bold>\n          <\/jats:p>","DOI":"10.1186\/s13068-025-02698-3","type":"journal-article","created":{"date-parts":[[2025,9,1]],"date-time":"2025-09-01T11:46:49Z","timestamp":1756727209000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Integrated co-cultivation and subsequent esterification: Harnessing Saccharomyces cerevisiae and Clostridium tyrobutyricum for streamlined ester production"],"prefix":"10.1186","volume":"18","author":[{"given":"Katharina","family":"Oehlenschl\u00e4ger","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michaela","family":"Lorenz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emily","family":"Schepp","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah","family":"Di Nonno","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dirk","family":"Holtmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roland","family":"Ulber","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,9,1]]},"reference":[{"key":"2698_CR1","doi-asserted-by":"publisher","DOI":"10.3109\/10242429809003198","author":"E Anderson","year":"1998","unstructured":"Anderson E, Larsson K, Kirk O. 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