{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T16:05:17Z","timestamp":1783008317119,"version":"3.54.5"},"reference-count":35,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,5,27]],"date-time":"2025-05-27T00:00:00Z","timestamp":1748304000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Croatian Science Foundation","award":["HRZZ-3075"],"award-info":[{"award-number":["HRZZ-3075"]}]},{"name":"Croatian Science Foundation","award":["2413\u20132024"],"award-info":[{"award-number":["2413\u20132024"]}]},{"name":"University of Zagreb institutional support of scientific activity","award":["HRZZ-3075"],"award-info":[{"award-number":["HRZZ-3075"]}]},{"name":"University of Zagreb institutional support of scientific activity","award":["2413\u20132024"],"award-info":[{"award-number":["2413\u20132024"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>Lactate is a valuable compound used in food, chemical, and pharmaceutical industries. High-value, optically pure L- or D-lactate can be synthesized microbially via specific dehydrogenases. The non-conventional yeast Scheffersomyces stipitis, which is known for fermenting both hexoses and pentoses, is a promising host for biochemical production from lignocellulosic biomass but does not naturally produce lactate. In this study, we engineered S. stipitis to produce lactate by expressing two codon-optimized bacterial L-lactate dehydrogenase genes under the control of strong native promoters. The engineered strain produced 7.42 g\/L (0.46 g\/g yield) and 11.67 g\/L (0.58 g\/g yield) lactate from glucose and xylose, respectively. The highest titer, 19.27 g\/L (0.52 g\/g yield), was achieved from 50 g\/L xylose after 74 h. Increasing the fermentation temperature from 28 \u00b0C to 32 \u00b0C improved yield by 30%, while a neutralizing agent further enhanced yield by 25% and prevented lactate degradation following carbon depletion. Although the wildtype strain produced a significant amount of ethanol on both glucose and xylose, the engineered strain produced ethanol as a side product exclusively on glucose and not on xylose. This phenomenon could be advantageous for biotechnological applications and may reflect shifts in gene expression depending on the carbon source or even on the presence of lactate.<\/jats:p>","DOI":"10.3390\/jof11060413","type":"journal-article","created":{"date-parts":[[2025,5,27]],"date-time":"2025-05-27T11:12:57Z","timestamp":1748344377000},"page":"413","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Production of Lactate by Metabolically Engineered Scheffersomyces stipitis"],"prefix":"10.3390","volume":"11","author":[{"given":"Angela","family":"Matanovi\u0107","sequence":"first","affiliation":[{"name":"Laboratory for Biology and Microbial Genetics, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3857-9818","authenticated-orcid":false,"given":"Nenad","family":"Mar\u0111etko","sequence":"additional","affiliation":[{"name":"Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-8296-5018","authenticated-orcid":false,"given":"Ana","family":"Sli\u0161kovi\u0107","sequence":"additional","affiliation":[{"name":"Laboratory for Biology and Microbial Genetics, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Blanka","family":"Didak","sequence":"additional","affiliation":[{"name":"Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5020-0639","authenticated-orcid":false,"given":"Karla","family":"Hanousek \u010ci\u010da","sequence":"additional","affiliation":[{"name":"Laboratory for Fermentation and Yeast Technology, Department of Food Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6855-7222","authenticated-orcid":false,"given":"Bojan","family":"\u017dunar","sequence":"additional","affiliation":[{"name":"Laboratory for Biology and Microbial Genetics, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anamarija","family":"\u0160tafa","sequence":"additional","affiliation":[{"name":"Laboratory for Biology and Microbial Genetics, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9583-534X","authenticated-orcid":false,"given":"Bo\u017eidar","family":"\u0160antek","sequence":"additional","affiliation":[{"name":"Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina","family":"Svetec Mikleni\u0107","sequence":"additional","affiliation":[{"name":"Laboratory for Biology and Microbial Genetics, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0469-2652","authenticated-orcid":false,"given":"Ivan-Kre\u0161imir","family":"Svetec","sequence":"additional","affiliation":[{"name":"Laboratory for Biology and Microbial Genetics, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1119","DOI":"10.1002\/jctb.1486","article-title":"Lactic acid: Recent advances in products, processes and technologies\u2014A review","volume":"81","author":"Datta","year":"2006","journal-title":"J. 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