fd9b6d1961f526b82f3a3b233940b5a7 FEMS.pdf a56bcca219299997b8356992749d69e8be68ea97 FEMS.pdf 0a136f6d1ffd2ddbfde29afa3c6b10fd1b88695a5409819bd64c4afe96c9fc41 FEMS.pdf Title: A native phosphoglycolate salvage pathway of the synthetic autotrophic yeast Komagataella phaffii Subject: DOI: 10.1093/femsml/uqad046 microLife, 5, , 11-12-2023. Abstract: Synthetic autotrophs can serve as chassis strains for bioproduction from CO2 as a feedstock to take measures against the climate crisis. Integration of the Calvin–Benson–Bassham (CBB) cycle into the methylotrophic yeast Komagataella phaffii (Pichia pastoris) enabled it to use CO2 as the sole carbon source. The key enzyme in this cycle is ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) catalyzing the carboxylation step. However, this enzyme is error prone to perform an oxygenation reaction leading to the production of toxic 2-phosphoglycolate. Native autotrophs have evolved different recycling pathways for 2-phosphoglycolate. However, for synthetic autotrophs, no information is available for the existence of such pathways. Deletion of CYB2 in the autotrophic K. phaffii strain led to the accumulation of glycolate, an intermediate in phosphoglycolate salvage pathways, suggesting that such a pathway is enabled by native K. phaffii enzymes. 13C tracer analysis with labeled glycolate indicated that the yeast pathway recycling phosphoglycolate is similar to the plant salvage pathway. This orthogonal yeast pathway may serve as a sensor for RuBisCO oxygenation, and as an engineering target to boost autotrophic growth rates in K. phaffii. Keywords: Author: Baumschabl Michael, Mitic Bernd M., Troyer Christina, Hann Stephan, Ata Özge, Mattanovich Diethard Creator: OUP Producer: Acrobat Distiller 23.0 (Windows); modified using iTextSharp 5.5.10 ©2000-2016 iText Group NV (AGPL-version) CreationDate: Tue Jan 16 11:05:17 2024 CET ModDate: Mon Apr 29 13:16:40 2024 CEST Custom Metadata: yes Metadata Stream: yes Tagged: yes UserProperties: no Suspects: no Form: none JavaScript: no Pages: 9 Encrypted: no Page size: 595.274 x 782.286 pts Page rot: 0 File size: 1792860 bytes Optimized: no PDF version: 1.4 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- IOGCNF+CaeciliaLTStd-Heavy Type 1C Custom yes yes yes 39 0 IOGCMF+MTSY Type 1C Custom yes yes yes 88 0 IOGCDH+CaeciliaLTStd-BoldItalic Type 1C Custom yes yes yes 127 0 IOGCCH+CaeciliaLTStd-Bold Type 1C Custom yes yes yes 40 0 IOGCCG+CaeciliaLTStd-Roman Type 1C Custom yes yes yes 309 0 IOGCCF+CaeciliaLTStd-Italic Type 1C Custom yes yes yes 41 0 IOGBKH+dummy-space Type 1C WinAnsi yes yes no 42 0 IOGBKG+CaeciliaLTStd-Light Type 1C Custom yes yes yes 43 0 UIULPV+ArialMT TrueType WinAnsi yes yes no 1 0 IOGECO+CaeciliaLTStd-LightItalic Type 1C Custom yes yes yes 38 0 IOGILD+RMTMI Type 1C Custom yes yes yes 128 0 IOGLCF+MTSYB Type 1C Custom yes yes yes 150 0 IOGLKD+MTEX Type 1C Custom yes yes no 177 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-05-03 02:40:29 CEST RepresentationInformation: FEMS.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-04-29 13:20:29 CEST Size: 1792860 Format: PDF Version: 1.4 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf Profile: Tagged PDF PDFMetadata: Objects: 1316 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: false CenterWindow: false DisplayDocTitle: true NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: OneColumn PageMode: UseOutlines Language: English Outlines: Item: Title: Introduction Destination: section.1.1.0 Item: Title: Results Destination: section.2.2.0 Item: Title: Material and methods Destination: section.3.7.0 Item: Title: Discussion Destination: section.4.15.0 Item: Title: Author contributions Destination: section.5.16.0 Item: Title: Acknowledgements Destination: section.6.17.0 Item: Title: Supplementary data Destination: section.7.18.0 Item: Title: Funding Destination: section.8.19.0 Item: Title: References Destination: section.9.19.0 Info: Title: A native phosphoglycolate salvage pathway of the synthetic autotrophic yeast Komagataella phaffii Author: Baumschabl Michael, Mitic Bernd M., Troyer Christina, Hann Stephan, Ata Ã zge, Mattanovich Diethard Subject: DOI: 10.1093/femsml/uqad046 microLife, 5, , 11-12-2023. Abstract: Synthetic autotrophs can serve as chassis strains for bioproduction from CO2 as a feedstock to take measures against the climate crisis. Integration of the Calvin–Benson–Bassham (CBB) cycle into the methylotrophic yeast Komagataella phaffii (Pichia pastoris) enabled it to use CO2 as the sole carbon source. The key enzyme in this cycle is ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) catalyzing the carboxylation step. However, this enzyme is error prone to perform an oxygenation reaction leading to the production of toxic 2-phosphoglycolate. Native autotrophs have evolved different recycling pathways for 2-phosphoglycolate. However, for synthetic autotrophs, no information is available for the existence of such pathways. Deletion of CYB2 in the autotrophic K. phaffii strain led to the accumulation of glycolate, an intermediate in phosphoglycolate salvage pathways, suggesting that such a pathway is enabled by native K. phaffii enzymes. 13C tracer analysis with labeled glycolate indicated that the yeast pathway recycling phosphoglycolate is similar to the plant salvage pathway. This orthogonal yeast pathway may serve as a sensor for RuBisCO oxygenation, and as an engineering target to boost autotrophic growth rates in K. phaffii. 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