Overexpression of the transcriptional activators Mxr1 and Mit1 enhances lactic acid production on methanol in Komagataella phaffii
Bachleitner, Simone (BOKU University. Department of Biotechnology)
Mølgaard Severinsen, Manja (BOKU University. Department of Biotechnology)
Lutz, Gregor (BOKU University. Department of Biotechnology)
Mattanovich, Diethard (BOKU University. Department of Biotechnology)

Fecha:	2024
Resumen:	A bio-based production of chemical building blocks from renewable, sustainable and non-food substrates is one key element to fight climate crisis. Lactic acid, one such chemical building block is currently produced from first generation feedstocks such as glucose and sucrose, both requiring land and water resources. In this study we aimed for lactic acid production from methanol by utilizing Komagataella phaffii as a production platform. Methanol, a single carbon source has potential as a sustainable substrate as technology allows (electro)chemical hydrogenation of CO2 for methanol production. Here we show that expression of the Lactiplantibacillus plantarum derived lactate dehydrogenase leads to L-lactic acid production in Komagataella phaffii, however, production resulted in low titers and cells subsequently consumed lactic acid again. Gene expression analysis of the methanol-utilizing genes AOX1, FDH1 and DAS2 showed that the presence of lactic acid downregulates transcription of the aforementioned genes, thereby repressing the methanol-utilizing pathway. For activation of the methanol-utilizing pathway in the presence of lactic acid, we constructed strains deficient in transcriptional repressors Nrg1, Mig1-1, and Mig1-2 as well as strains with overrepresentation of transcriptional activators Mxr1 and Mit1. While loss of transcriptional repressors had no significant impact on lactic acid production, overexpression of both transcriptional activators, MXR1 and MIT1, increased lactic acid titers from 4 g L-1 to 17 g L-1 in bioreactor cultivations.
Ayudas:	European Commission 101000441
Derechos:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió publicada
Materia:	Lactic acid; ; Komagataella phaffii; ; Transcriptional regulation; ; Methanol; ; Synthetic biology; ; Circular biotechnology
Publicado en:	Metabolic Engineering, Vol. 85 (August 2024) , pag. 133-144, ISSN 1096-7184

DOI: 10.1016/j.ymben.2024.07.013

12 p, 1.2 MB

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