{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T15:24:16Z","timestamp":1789658656484,"version":"4.0.1"},"reference-count":56,"publisher":"Springer Science and Business Media LLC","issue":"9","license":[{"start":{"date-parts":[[2026,8,28]],"date-time":"2026-08-28T00:00:00Z","timestamp":1787875200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,8,28]],"date-time":"2026-08-28T00:00:00Z","timestamp":1787875200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"Forschungszentrum J\u00fclich GmbH","id":[{"id":"https:\/\/ror.org\/02nv7yv05","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Bioprocess Biosyst Eng"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    The replacement of cobalt-based siccatives in unsaturated polyester resin curing requires sustainable and scalable biocatalytic alternatives, with fungal peroxidases representing a promising option. For industrial implementation, however, robust large-scale production processes are essential. In this study, we investigated the robustness of a glycerol-based peroxidase production process in\n                    <jats:italic>K. phaffii<\/jats:italic>\n                    driven by the glycerol-derepressible P\n                    <jats:sub>DF<\/jats:sub>\n                    promoter, focusing on industrially relevant bioreactor inhomogeneities. A two-compartment scale-down system was used to mimic substrate and oxygen gradients arising from insufficient mixing and mass transfer at large scale. Substrate inhomogeneity alone led to a moderate reduction in peroxidase titers (approx. 23%), indicating partial sensitivity to localized substrate oversupply. In contrast, simultaneous substrate and oxygen inhomogeneity resulted in a counterintuitive increase in peroxidase titers of approx. 13% compared to homogeneous reference conditions. This suggests that dynamic transitions between substrate limitation and surplus can positively affect peroxidase formation under P\n                    <jats:sub>DF<\/jats:sub>\n                    promoter control. Based on this observation, a modified process strategy incorporating glycerol pulses during the depression phase was evaluated. While this approach demonstrated high robustness, it did not enhance productivity. Overall, the results demonstrate that the developed\n                    <jats:italic>K. phaffii<\/jats:italic>\n                    process is resilient to large-scale inhomogeneities and can even benefit from specific dynamic cultivation conditions.\n                  <\/jats:p>","DOI":"10.1007\/s00449-026-03415-6","type":"journal-article","created":{"date-parts":[[2026,8,28]],"date-time":"2026-08-28T04:41:09Z","timestamp":1787892069000},"page":"2429-2440","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Thinking big in small scale: characterization of peroxidase production in Komagataella phaffii using two-compartment scale-down system"],"prefix":"10.1007","volume":"49","author":[{"given":"Christian Mathis","family":"Wagner","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katharina","family":"Schwartz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anton","family":"Glieder","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wolfgang","family":"Wiechert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0704-5597","authenticated-orcid":false,"given":"Marco","family":"Oldiges","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,8,28]]},"reference":[{"key":"3415_CR1","doi-asserted-by":"publisher","first-page":"198","DOI":"10.1016\/j.porgcoat.2005.06.004","volume":"54","author":"ZO Oyman","year":"2005","unstructured":"Oyman ZO, Ming W, Linde RVD (2005) Oxidation of drying oils containing non-conjugated and conjugated double bonds catalyzed by a cobalt catalyst. Prog Org Coat 54:198\u2013204. https:\/\/doi.org\/10.1016\/j.porgcoat.2005.06.004","journal-title":"Prog Org Coat"},{"key":"3415_CR2","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1016\/S0034-3617(13)70056-0","volume":"57","author":"R Zuijderduin","year":"2013","unstructured":"Zuijderduin R (2013) Cobalt-free curing based on iron. Reinf Plast 57:25\u201328. https:\/\/doi.org\/10.1016\/S0034-3617(13)70056-0","journal-title":"Reinf Plast"},{"key":"3415_CR3","unstructured":"(2023) Cobalt Market Report. Cobalt Institute"},{"key":"3415_CR4","doi-asserted-by":"publisher","first-page":"427","DOI":"10.3109\/10408449009029330","volume":"20","author":"AA Jensen","year":"1990","unstructured":"Jensen AA, Tuchsen F (1990) Cobalt Exposure and Cancer Risk. Crit Rev Toxicol 20:427\u2013439. https:\/\/doi.org\/10.3109\/10408449009029330","journal-title":"Crit Rev Toxicol"},{"key":"3415_CR5","doi-asserted-by":"publisher","first-page":"577","DOI":"10.1016\/S1470-2045(22)00219-4","volume":"23","author":"MR Karagas","year":"2022","unstructured":"Karagas MR, Wang A, Dorman DC et al (2022) Carcinogenicity of cobalt, antimony compounds, and weapons-grade tungsten alloy. Lancet Oncol 23:577\u2013578. https:\/\/doi.org\/10.1016\/S1470-2045(22)00219-4","journal-title":"Lancet Oncol"},{"key":"3415_CR6","doi-asserted-by":"publisher","first-page":"142","DOI":"10.1002\/masy.201200102","volume":"329","author":"JFGA Jansen","year":"2013","unstructured":"Jansen JFGA, Hilker I, Kleuskens E et al (2013) Cobalt Replacement in Unsaturated Polyester Resins; Going for Sustainable Composites. Macromol Symp 329:142\u2013149. https:\/\/doi.org\/10.1002\/masy.201200102","journal-title":"Macromol Symp"},{"key":"3415_CR7","doi-asserted-by":"publisher","first-page":"1631","DOI":"10.1111\/jiec.13258","volume":"26","author":"M Van Der Meide","year":"2022","unstructured":"Van Der Meide M, Harpprecht C, Northey S et al (2022) Effects of the energy transition on environmental impacts of cobalt supply: A prospective life cycle assessment study on future supply of cobalt. J Ind Ecol 26:1631\u20131645. https:\/\/doi.org\/10.1111\/jiec.13258","journal-title":"J Ind Ecol"},{"key":"3415_CR8","doi-asserted-by":"publisher","first-page":"107615","DOI":"10.1016\/j.biotechadv.2020.107615","volume":"51","author":"M Hobisch","year":"2021","unstructured":"Hobisch M, Holtmann D, Gomez De Santos P et al (2021) Recent developments in the use of peroxygenases \u2013 Exploring their high potential in selective oxyfunctionalisations. Biotechnol Adv 51:107615. https:\/\/doi.org\/10.1016\/j.biotechadv.2020.107615","journal-title":"Biotechnol Adv"},{"key":"3415_CR9","doi-asserted-by":"publisher","first-page":"5740","DOI":"10.1007\/s12010-023-04835-w","volume":"196","author":"C Shen","year":"2024","unstructured":"Shen C, Wang Y (2024) Recent Progress on Peroxidase Modification and Application. Appl Biochem Biotechnol 196:5740\u20135764. https:\/\/doi.org\/10.1007\/s12010-023-04835-w","journal-title":"Appl Biochem Biotechnol"},{"key":"3415_CR10","unstructured":"Radespiel T, Borup B, Thielmann J, Biecker C (2022) Curable Composition"},{"key":"3415_CR11","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1016\/S0168-1656(01)00394-7","volume":"93","author":"A Conesa","year":"2002","unstructured":"Conesa A, Punt PJ, Van Den Hondel CAMJJ (2002) Fungal peroxidases: molecular aspects and applications. J Biotechnol 93:143\u2013158. https:\/\/doi.org\/10.1016\/S0168-1656(01)00394-7","journal-title":"J Biotechnol"},{"key":"3415_CR12","doi-asserted-by":"publisher","first-page":"349","DOI":"10.1016\/j.pbi.2008.02.003","volume":"11","author":"K Hammel","year":"2008","unstructured":"Hammel K, Cullen D (2008) Role of fungal peroxidases in biological ligninolysis. Curr Opin Plant Biol 11:349\u2013355. https:\/\/doi.org\/10.1016\/j.pbi.2008.02.003","journal-title":"Curr Opin Plant Biol"},{"key":"3415_CR13","doi-asserted-by":"publisher","first-page":"116","DOI":"10.1016\/j.cbpa.2014.01.015","volume":"19","author":"M Hofrichter","year":"2014","unstructured":"Hofrichter M, Ullrich R (2014) Oxidations catalyzed by fungal peroxygenases. Curr Opin Chem Biol 19:116\u2013125. https:\/\/doi.org\/10.1016\/j.cbpa.2014.01.015","journal-title":"Curr Opin Chem Biol"},{"key":"3415_CR14","doi-asserted-by":"publisher","first-page":"185","DOI":"10.1016\/S1381-1177(98)00119-2","volume":"6","author":"T Nakayama","year":"1999","unstructured":"Nakayama T, Amachi T (1999) Fungal peroxidase: its structure, function, and application. J Mol Catal B Enzym 6:185\u2013198. https:\/\/doi.org\/10.1016\/S1381-1177(98)00119-2","journal-title":"J Mol Catal B Enzym"},{"key":"3415_CR15","doi-asserted-by":"publisher","first-page":"11511","DOI":"10.1021\/acscatal.1c03065","volume":"11","author":"L Rotilio","year":"2021","unstructured":"Rotilio L, Swoboda A, Ebner K et al (2021) Structural and Biochemical Studies Enlighten the Unspecific Peroxygenase from Hypoxylon sp. EC38 as an Efficient Oxidative Biocatalyst. ACS Catal 11:11511\u201311525. https:\/\/doi.org\/10.1021\/acscatal.1c03065","journal-title":"ACS Catal"},{"key":"3415_CR16","doi-asserted-by":"publisher","first-page":"441","DOI":"10.3390\/biom13030441","volume":"13","author":"GD Barone","year":"2023","unstructured":"Barone GD, Emmerstorfer-Augustin A, Biundo A et al (2023) Industrial Production of Proteins with Pichia pastoris\u2014Komagataella phaffii. Biomolecules 13:441. https:\/\/doi.org\/10.3390\/biom13030441","journal-title":"Biomolecules"},{"key":"3415_CR17","doi-asserted-by":"publisher","first-page":"346","DOI":"10.3390\/microorganisms12020346","volume":"12","author":"TM Khlebodarova","year":"2024","unstructured":"Khlebodarova TM, Bogacheva NV, Zadorozhny AV et al (2024) Komagataella phaffii as a Platform for Heterologous Expression of Enzymes Used for Industry. Microorganisms 12:346. https:\/\/doi.org\/10.3390\/microorganisms12020346","journal-title":"Microorganisms"},{"key":"3415_CR18","doi-asserted-by":"publisher","first-page":"607028","DOI":"10.3389\/fmicb.2020.607028","volume":"11","author":"L Bernauer","year":"2021","unstructured":"Bernauer L, Radkohl A, Lehmayer LGK, Emmerstorfer-Augustin A (2021) Komagataella phaffii as Emerging Model Organism in Fundamental Research. Front Microbiol 11:607028. https:\/\/doi.org\/10.3389\/fmicb.2020.607028","journal-title":"Front Microbiol"},{"key":"3415_CR19","doi-asserted-by":"publisher","first-page":"346","DOI":"10.3390\/metabo12040346","volume":"12","author":"C Bustos","year":"2022","unstructured":"Bustos C, Quezada J, Veas R et al (2022) Advances in Cell Engineering of the Komagataella phaffii Platform for Recombinant Protein Production. Metabolites 12:346. https:\/\/doi.org\/10.3390\/metabo12040346","journal-title":"Metabolites"},{"key":"3415_CR20","doi-asserted-by":"publisher","first-page":"22","DOI":"10.1186\/1475-2859-11-22","volume":"11","author":"FW Krainer","year":"2012","unstructured":"Krainer FW, Dietzsch C, Hajek T et al (2012) Recombinant protein expression in Pichia pastoris strains with an engineered methanol utilization pathway. Microb Cell Factories 11:22. https:\/\/doi.org\/10.1186\/1475-2859-11-22","journal-title":"Microb Cell Factories"},{"key":"3415_CR21","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1186\/s12934-015-0292-7","volume":"14","author":"S Capone","year":"2015","unstructured":"Capone S, Horvat J, Herwig C, Spadiut O (2015) Development of a mixed feed strategy for a recombinant Pichia pastoris strain producing with a de-repression promoter. Microb Cell Factories 14:101. https:\/\/doi.org\/10.1186\/s12934-015-0292-7","journal-title":"Microb Cell Factories"},{"key":"3415_CR22","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1111\/j.1574-6976.2000.tb00532.x","volume":"24","author":"JL Cereghino","year":"2000","unstructured":"Cereghino JL, Cregg JM (2000) Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol Rev 24:45\u201366. https:\/\/doi.org\/10.1111\/j.1574-6976.2000.tb00532.x","journal-title":"FEMS Microbiol Rev"},{"key":"3415_CR23","doi-asserted-by":"publisher","first-page":"385","DOI":"10.1016\/j.nbt.2012.11.010","volume":"30","author":"T Vogl","year":"2013","unstructured":"Vogl T, Glieder A (2013) Regulation of Pichia pastoris promoters and its consequences for protein production. New Biotechnol 30:385\u2013404. https:\/\/doi.org\/10.1016\/j.nbt.2012.11.010","journal-title":"New Biotechnol"},{"key":"3415_CR24","doi-asserted-by":"publisher","DOI":"10.3791\/58589","author":"JE Fischer","year":"2019","unstructured":"Fischer JE, Hatzl A-M, Weninger A et al (2019) Methanol independent expression by Pichia pastoris employing de-repression technologies. J Vis Exp. https:\/\/doi.org\/10.3791\/58589","journal-title":"J Vis Exp"},{"key":"3415_CR25","doi-asserted-by":"publisher","first-page":"38","DOI":"10.1186\/s13568-020-00972-1","volume":"10","author":"T Vogl","year":"2020","unstructured":"Vogl T, Fischer JE, Hyden P et al (2020) Orthologous promoters from related methylotrophic yeasts surpass expression of endogenous promoters of Pichia pastoris. AMB Express 10:38. https:\/\/doi.org\/10.1186\/s13568-020-00972-1","journal-title":"AMB Express"},{"key":"3415_CR26","doi-asserted-by":"publisher","first-page":"891","DOI":"10.1007\/s13213-013-0767-x","volume":"64","author":"D Samul","year":"2014","unstructured":"Samul D, Leja K, Grajek W (2014) Impurities of crude glycerol and their effect on metabolite production. Ann Microbiol 64:891\u2013898. https:\/\/doi.org\/10.1007\/s13213-013-0767-x","journal-title":"Ann Microbiol"},{"key":"3415_CR27","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1186\/1754-6834-5-13","volume":"5","author":"F Yang","year":"2012","unstructured":"Yang F, Hanna MA, Sun R (2012) Value-added uses for crude glycerol\u2013a byproduct of biodiesel production. Biotechnol Biofuels 5:13. https:\/\/doi.org\/10.1186\/1754-6834-5-13","journal-title":"Biotechnol Biofuels"},{"key":"3415_CR28","first-page":"1","volume-title":"Process Control, Intensification, and Digitalisation in Continuous Biomanufacturing","author":"J Kopp","year":"2022","unstructured":"Kopp J, Slouka C, Delvigne F, Herwig C (2022) Strategies for Continuous Processing in Microbial Systems. In: Subramanian G (ed) Process Control, Intensification, and Digitalisation in Continuous Biomanufacturing, 1st edn. Wiley, pp 1\u201337","edition":"1st edn"},{"key":"3415_CR29","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1186\/1475-2859-12-5","volume":"12","author":"R Prielhofer","year":"2013","unstructured":"Prielhofer R, Maurer M, Klein J et al (2013) Induction without methanol: novel regulated promoters enable high-level expression in Pichia pastoris. Microb Cell Factories 12:5. https:\/\/doi.org\/10.1186\/1475-2859-12-5","journal-title":"Microb Cell Factories"},{"key":"3415_CR30","doi-asserted-by":"publisher","first-page":"36","DOI":"10.1016\/j.ymben.2022.08.010","volume":"74","author":"RJ Zahrl","year":"2022","unstructured":"Zahrl RJ, Prielhofer R, Ata \u00d6 et al (2022) Pushing and pulling proteins into the yeast secretory pathway enhances recombinant protein secretion. Metab Eng 74:36\u201348. https:\/\/doi.org\/10.1016\/j.ymben.2022.08.010","journal-title":"Metab Eng"},{"key":"3415_CR31","doi-asserted-by":"publisher","first-page":"114","DOI":"10.1016\/j.copbio.2010.02.001","volume":"21","author":"P Neubauer","year":"2010","unstructured":"Neubauer P, Junne S (2010) Scale-down simulators for metabolic analysis of large-scale bioprocesses. Curr Opin Biotechnol 21:114\u2013121. https:\/\/doi.org\/10.1016\/j.copbio.2010.02.001","journal-title":"Curr Opin Biotechnol"},{"key":"3415_CR32","doi-asserted-by":"publisher","first-page":"425","DOI":"10.1007\/s00253-005-0003-0","volume":"68","author":"FR Schmidt","year":"2005","unstructured":"Schmidt FR (2005) Optimization and scale up of industrial fermentation processes. Appl Microbiol Biotechnol 68:425\u2013435. https:\/\/doi.org\/10.1007\/s00253-005-0003-0","journal-title":"Appl Microbiol Biotechnol"},{"key":"3415_CR33","doi-asserted-by":"publisher","first-page":"546","DOI":"10.1002\/bit.260260522","volume":"26","author":"NMG Oosterhuis","year":"1984","unstructured":"Oosterhuis NMG, Kossen NWF (1984) Dissolved oxygen concentration profiles in a production-scale bioreactor. Biotechnol Bioeng 26:546\u2013550. https:\/\/doi.org\/10.1002\/bit.260260522","journal-title":"Biotechnol Bioeng"},{"key":"3415_CR34","doi-asserted-by":"publisher","first-page":"3194","DOI":"10.1002\/bit.28208","volume":"119","author":"S T\u00e4uber","year":"2022","unstructured":"T\u00e4uber S, Bl\u00f6baum L, Steier V et al (2022) Microfluidic single-cell scale\u2010down bioreactors: A proof\u2010of\u2010concept for the growth of Corynebacterium glutamicum at oscillating pH values. Biotechnol Bioeng 119:3194\u20133209. https:\/\/doi.org\/10.1002\/bit.28208","journal-title":"Biotechnol Bioeng"},{"key":"3415_CR35","doi-asserted-by":"publisher","first-page":"560","DOI":"10.1002\/bit.26184","volume":"114","author":"MH Limberg","year":"2017","unstructured":"Limberg MH, Schulte J, Aryani T et al (2017) Metabolic profile of 1,5-diaminopentane producing Corynebacterium glutamicum under scale\u2010down conditions: Blueprint for robustness to bioreactor inhomogeneities. Biotechnol Bioeng 114:560\u2013575. https:\/\/doi.org\/10.1002\/bit.26184","journal-title":"Biotechnol Bioeng"},{"key":"3415_CR36","unstructured":"Invitrogen AG (2002) Pichia Fermentation Process Guidelines"},{"key":"3415_CR37","doi-asserted-by":"publisher","first-page":"610","DOI":"10.1002\/elsc.201500142","volume":"16","author":"MH Limberg","year":"2016","unstructured":"Limberg MH, Pooth V, Wiechert W, Oldiges M (2016) Plug flow versus stirred tank reactor flow characteristics in two-compartment scale\u2010down bioreactor: Setup\u2010specific influence on the metabolic phenotype and bioprocess performance of Corynebacterium glutamicum. Eng Life Sci 16:610\u2013619. https:\/\/doi.org\/10.1002\/elsc.201500142","journal-title":"Eng Life Sci"},{"key":"3415_CR38","doi-asserted-by":"publisher","first-page":"1619","DOI":"10.1007\/s00449-025-03182-w","volume":"48","author":"P Arulrajah","year":"2025","unstructured":"Arulrajah P, Lievonen AE, Suba\u015f\u0131 D et al (2025) Scale-down bioreactors\u2014comparative analysis of configurations. Bioprocess Biosyst Eng 48:1619\u20131635. https:\/\/doi.org\/10.1007\/s00449-025-03182-w","journal-title":"Bioprocess Biosyst Eng"},{"key":"3415_CR39","doi-asserted-by":"publisher","first-page":"1009","DOI":"10.1002\/biot.201100293","volume":"6","author":"S Junne","year":"2011","unstructured":"Junne S, Klingner A, Kabisch J et al (2011) A two-compartment bioreactor system made of commercial parts for bioprocess scale\u2010down studies: Impact of oscillations on Bacillus subtilis fed\u2010batch cultivations. Biotechnol J 6:1009\u20131017. https:\/\/doi.org\/10.1002\/biot.201100293","journal-title":"Biotechnol J"},{"key":"3415_CR40","doi-asserted-by":"publisher","first-page":"567","DOI":"10.1002\/bit.260310609","volume":"31","author":"APJ Sweere","year":"1988","unstructured":"Sweere APJ, Mesters JR, Janse L et al (1988) Experimental simulation of oxygen profiles and their influence on baker\u2019s yeast production: I. One-fermentor system. Biotechnol Bioeng 31:567\u2013578. https:\/\/doi.org\/10.1002\/bit.260310609","journal-title":"Biotechnol Bioeng"},{"key":"3415_CR41","doi-asserted-by":"publisher","first-page":"249","DOI":"10.1007\/BF01061530","volume":"9","author":"S George","year":"1993","unstructured":"George S, Larsson G, Enfors S-O (1993) A scale-down two-compartment reactor with controlled substrate oscillations: Metabolic response of Saccharomyces cerevisiae. Bioprocess Eng 9:249\u2013257. https:\/\/doi.org\/10.1007\/BF01061530","journal-title":"Bioprocess Eng"},{"key":"3415_CR42","doi-asserted-by":"publisher","first-page":"453","DOI":"10.1002\/bit.20383","volume":"89","author":"EA Sandoval-Basurto","year":"2005","unstructured":"Sandoval-Basurto EA, Gosset G, Bol\u00edvar F, Ram\u00edrez OT (2005) Culture of Escherichia coli under dissolved oxygen gradients simulated in a two\u2010compartment scale\u2010down system: Metabolic response and production of recombinant protein. Biotechnol Bioeng 89:453\u2013463. https:\/\/doi.org\/10.1002\/bit.20383","journal-title":"Biotechnol Bioeng"},{"key":"3415_CR43","doi-asserted-by":"publisher","first-page":"152","DOI":"10.1002\/btpr.2184","volume":"32","author":"N Risager Wright","year":"2016","unstructured":"Risager Wright N, R\u00f8nnest NP, Thykaer J (2016) Scale-down of continuous protein producing Saccharomyces cerevisiae cultivations using a two\u2010compartment system. Biotechnol Prog 32:152\u2013159. https:\/\/doi.org\/10.1002\/btpr.2184","journal-title":"Biotechnol Prog"},{"key":"3415_CR44","doi-asserted-by":"publisher","first-page":"52","DOI":"10.1016\/j.nbt.2019.01.005","volume":"50","author":"M Tom\u00e0s-Gamisans","year":"2019","unstructured":"Tom\u00e0s-Gamisans M, \u00d8dum ASR, Workman M et al (2019) Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysis. New Biotechnol 50:52\u201359. https:\/\/doi.org\/10.1016\/j.nbt.2019.01.005","journal-title":"New Biotechnol"},{"key":"3415_CR45","doi-asserted-by":"publisher","first-page":"65","DOI":"10.1186\/1475-2859-8-65","volume":"8","author":"M Carnicer","year":"2009","unstructured":"Carnicer M, Baumann K, T\u00f6plitz I et al (2009) Macromolecular and elemental composition analysis and extracellular metabolite balances of Pichia pastoris growing at different oxygen levels. Microb Cell Factories 8:65. https:\/\/doi.org\/10.1186\/1475-2859-8-65","journal-title":"Microb Cell Factories"},{"key":"3415_CR46","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1186\/1752-0509-4-141","volume":"4","author":"K Baumann","year":"2010","unstructured":"Baumann K, Carnicer M, Dragosits M et al (2010) A multi-level study of recombinant Pichia pastoris in different oxygen conditions. BMC Syst Biol 4:141. https:\/\/doi.org\/10.1186\/1752-0509-4-141","journal-title":"BMC Syst Biol"},{"key":"3415_CR47","doi-asserted-by":"publisher","first-page":"177","DOI":"10.1002\/bit.21763","volume":"100","author":"K Baumann","year":"2008","unstructured":"Baumann K, Maurer M, Dragosits M et al (2008) Hypoxic fed-batch cultivation of Pichia pastoris increases specific and volumetric productivity of recombinant proteins. Biotechnol Bioeng 100:177\u2013183. https:\/\/doi.org\/10.1002\/bit.21763","journal-title":"Biotechnol Bioeng"},{"key":"3415_CR48","doi-asserted-by":"publisher","first-page":"e14411","DOI":"10.1111\/1751-7915.14411","volume":"17","author":"A Sales-Vallverd\u00fa","year":"2024","unstructured":"Sales-Vallverd\u00fa A, Gasset A, Requena\u2010Moreno G et al (2024) Synergic kinetic and physiological control to improve the efficiency of Komagataella phaffii recombinant protein production bioprocesses. Microb Biotechnol 17:e14411. https:\/\/doi.org\/10.1111\/1751-7915.14411","journal-title":"Microb Biotechnol"},{"key":"3415_CR49","doi-asserted-by":"publisher","first-page":"74","DOI":"10.1186\/s12934-021-01564-9","volume":"20","author":"J Garrig\u00f3s-Mart\u00ednez","year":"2021","unstructured":"Garrig\u00f3s-Mart\u00ednez J, Vuoristo K, Nieto-Taype MA et al (2021) Bioprocess performance analysis of novel methanol-independent promoters for recombinant protein production with Pichia pastoris. Microb Cell Factories 20:74. https:\/\/doi.org\/10.1186\/s12934-021-01564-9","journal-title":"Microb Cell Factories"},{"key":"3415_CR50","doi-asserted-by":"publisher","first-page":"357","DOI":"10.1002\/bit.22836","volume":"107","author":"J Heyland","year":"2010","unstructured":"Heyland J, Fu J, Blank LM, Schmid A (2010) Quantitative physiology of Pichia pastoris during glucose-limited high\u2010cell density fed\u2010batch cultivation for recombinant protein production. Biotechnol Bioeng 107:357\u2013368. https:\/\/doi.org\/10.1002\/bit.22836","journal-title":"Biotechnol Bioeng"},{"key":"3415_CR51","doi-asserted-by":"publisher","first-page":"1942","DOI":"10.1002\/bit.23114","volume":"108","author":"J Heyland","year":"2011","unstructured":"Heyland J, Fu J, Blank LM, Schmid A (2011) Carbon metabolism limits recombinant protein production in Pichia pastoris. Biotechnol Bioeng 108:1942\u20131953. https:\/\/doi.org\/10.1002\/bit.23114","journal-title":"Biotechnol Bioeng"},{"key":"3415_CR52","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1186\/1475-2859-11-57","volume":"11","author":"J Jord\u00e0","year":"2012","unstructured":"Jord\u00e0 J, Jouhten P, C\u00e1mara E et al (2012) Metabolic flux profiling of recombinant protein secreting Pichia pastoris growing on glucose:methanol mixtures. Microb Cell Factories 11:57. https:\/\/doi.org\/10.1186\/1475-2859-11-57","journal-title":"Microb Cell Factories"},{"key":"3415_CR53","doi-asserted-by":"publisher","first-page":"3163","DOI":"10.1007\/s00253-017-8123-x","volume":"101","author":"V Looser","year":"2017","unstructured":"Looser V, L\u00fcthy D, Straumann M et al (2017) Effects of glycerol supply and specific growth rate on methanol-free production of CALB by P. pastoris: functional characterisation of a novel promoter. Appl Microbiol Biotechnol 101:3163\u20133176. https:\/\/doi.org\/10.1007\/s00253-017-8123-x","journal-title":"Appl Microbiol Biotechnol"},{"key":"3415_CR54","doi-asserted-by":"publisher","first-page":"367","DOI":"10.1002\/bit.10904","volume":"85","author":"H Hohenblum","year":"2004","unstructured":"Hohenblum H, Gasser B, Maurer M et al (2004) Effects of gene dosage, promoters, and substrates on unfolded protein stress of recombinant Pichia pastoris. Biotechnol Bioeng 85:367\u2013375. https:\/\/doi.org\/10.1002\/bit.10904","journal-title":"Biotechnol Bioeng"},{"key":"3415_CR55","doi-asserted-by":"publisher","first-page":"21","DOI":"10.1186\/1475-2859-6-21","volume":"6","author":"D Resina","year":"2007","unstructured":"Resina D, Boll\u00f3k M, Khatri NK et al (2007) Transcriptional response of P. pastoris in fed-batch cultivations to Rhizopus oryzae lipase production reveals UPR induction. Microb Cell Factories 6:21. https:\/\/doi.org\/10.1186\/1475-2859-6-21","journal-title":"Microb Cell Factories"},{"key":"3415_CR56","doi-asserted-by":"publisher","DOI":"10.1186\/s12934-026-03055-1","author":"CM Wagner","year":"2026","unstructured":"Wagner CM, Schwartz K, Czech H et al (2026) Development and optimization of a glycerol-based fed-batch strategy for the production of peroxidases with Komagataella phaffii under the PDF promoter. Microb Cell Factories. https:\/\/doi.org\/10.1186\/s12934-026-03055-1","journal-title":"Microb Cell Factories"}],"container-title":["Bioprocess and Biosystems Engineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00449-026-03415-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s00449-026-03415-6","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00449-026-03415-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T09:01:39Z","timestamp":1789549299000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s00449-026-03415-6"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,28]]},"references-count":56,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2026,9]]}},"alternative-id":["3415"],"URL":"https:\/\/doi.org\/10.1007\/s00449-026-03415-6","relation":{},"ISSN":["1615-7591","1615-7605"],"issn-type":[{"value":"1615-7591","type":"print"},{"value":"1615-7605","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,28]]},"assertion":[{"value":"17 March 2026","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"13 August 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"28 August 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","label":"Conflict of interest","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"All co-authors have given their consent to publish this manuscript.","order":2,"name":"Ethics","label":"Consent for publication","group":{"name":"EthicsHeading","label":"Declarations"}}]}}