Role of Dissimilative Pathway of <i>Komagataella phaffii</i> (<i>Pichia pastoris</i>): Formaldehyde Toxicity and Energy Metabolism
<i>Komagataella phaffii</i> (aka <i>Pichia pastoris</i>) is a yeast able to grow in methanol as the sole carbon and energy source. This substrate is converted into formaldehyde, a toxic intermediary that can either be assimilated to biomass or dissimilated to CO<sub>2&l...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-07-01
|
| Series: | Microorganisms |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2607/10/7/1466 |
| _version_ | 1827618791653113856 |
|---|---|
| author | Julio Berrios Chrispian W. Theron Sébastien Steels Belén Ponce Edgar Velastegui Cristina Bustos Claudia Altamirano Patrick Fickers |
| author_facet | Julio Berrios Chrispian W. Theron Sébastien Steels Belén Ponce Edgar Velastegui Cristina Bustos Claudia Altamirano Patrick Fickers |
| author_sort | Julio Berrios |
| collection | DOAJ |
| description | <i>Komagataella phaffii</i> (aka <i>Pichia pastoris</i>) is a yeast able to grow in methanol as the sole carbon and energy source. This substrate is converted into formaldehyde, a toxic intermediary that can either be assimilated to biomass or dissimilated to CO<sub>2</sub> through the enzymes formaldehyde dehydrogenase (FLD) and formate dehydrogenase, also producing energy in the form of NADH. The dissimilative pathway has been described as an energy producing and a detoxifying route, but conclusive evidence has not been provided for this. In order to elucidate this theory, we generated mutants lacking the FLD activity (Δ<i>fld1</i>) and used flux analysis to evaluate the metabolic impact of this disrupted pathway. Unexpectedly, we found that the specific growth rate of the Δ<i>fld1</i> strain was only slightly lower (92%) than the control. In contrast, the sensitivity to formaldehyde pulses (up to 8mM) was significantly higher in the Δ<i>fld1</i> mutant strain and was associated with a higher maintenance energy. In addition, the intracellular flux estimation revealed a high metabolic flexibility of <i>K. phaffii</i> in response to the disrupted pathway. Our results suggest that the role of the dissimilative pathway is mainly to protect the cells from the harmful effect of formaldehyde, as they were able to compensate for the energy provided from this pathway when disrupted. |
| first_indexed | 2024-03-09T10:14:21Z |
| format | Article |
| id | doaj.art-61dc359359c9416c829c7020c45c68bc |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-09T10:14:21Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-61dc359359c9416c829c7020c45c68bc2023-12-01T22:28:48ZengMDPI AGMicroorganisms2076-26072022-07-01107146610.3390/microorganisms10071466Role of Dissimilative Pathway of <i>Komagataella phaffii</i> (<i>Pichia pastoris</i>): Formaldehyde Toxicity and Energy MetabolismJulio Berrios0Chrispian W. Theron1Sébastien Steels2Belén Ponce3Edgar Velastegui4Cristina Bustos5Claudia Altamirano6Patrick Fickers7School of Biochemical Engineering, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2085, Valparaíso 2340000, ChileGeneMill, Institute of Systems, Molecular and Integrative Biology, Biosciences Building, University of Liverpool, Crown Street, Liverpool L69 7BE, UKMicrobial Processes and Interactions, TERRA Teaching and Research Centre, Gembloux Agro-Bio Tech, University of Liège, Av. de la Faculté 2B, 5030 Gembloux, BelgiumSchool of Biochemical Engineering, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2085, Valparaíso 2340000, ChileSchool of Biochemical Engineering, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2085, Valparaíso 2340000, ChileSchool of Biochemical Engineering, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2085, Valparaíso 2340000, ChileSchool of Biochemical Engineering, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2085, Valparaíso 2340000, ChileMicrobial Processes and Interactions, TERRA Teaching and Research Centre, Gembloux Agro-Bio Tech, University of Liège, Av. de la Faculté 2B, 5030 Gembloux, Belgium<i>Komagataella phaffii</i> (aka <i>Pichia pastoris</i>) is a yeast able to grow in methanol as the sole carbon and energy source. This substrate is converted into formaldehyde, a toxic intermediary that can either be assimilated to biomass or dissimilated to CO<sub>2</sub> through the enzymes formaldehyde dehydrogenase (FLD) and formate dehydrogenase, also producing energy in the form of NADH. The dissimilative pathway has been described as an energy producing and a detoxifying route, but conclusive evidence has not been provided for this. In order to elucidate this theory, we generated mutants lacking the FLD activity (Δ<i>fld1</i>) and used flux analysis to evaluate the metabolic impact of this disrupted pathway. Unexpectedly, we found that the specific growth rate of the Δ<i>fld1</i> strain was only slightly lower (92%) than the control. In contrast, the sensitivity to formaldehyde pulses (up to 8mM) was significantly higher in the Δ<i>fld1</i> mutant strain and was associated with a higher maintenance energy. In addition, the intracellular flux estimation revealed a high metabolic flexibility of <i>K. phaffii</i> in response to the disrupted pathway. Our results suggest that the role of the dissimilative pathway is mainly to protect the cells from the harmful effect of formaldehyde, as they were able to compensate for the energy provided from this pathway when disrupted.https://www.mdpi.com/2076-2607/10/7/1466<i>Komagataella phaffii</i><i>Pichia pastoris</i>methanolformaldehyde dehydrogenasedissimilative pathway |
| spellingShingle | Julio Berrios Chrispian W. Theron Sébastien Steels Belén Ponce Edgar Velastegui Cristina Bustos Claudia Altamirano Patrick Fickers Role of Dissimilative Pathway of <i>Komagataella phaffii</i> (<i>Pichia pastoris</i>): Formaldehyde Toxicity and Energy Metabolism Microorganisms <i>Komagataella phaffii</i> <i>Pichia pastoris</i> methanol formaldehyde dehydrogenase dissimilative pathway |
| title | Role of Dissimilative Pathway of <i>Komagataella phaffii</i> (<i>Pichia pastoris</i>): Formaldehyde Toxicity and Energy Metabolism |
| title_full | Role of Dissimilative Pathway of <i>Komagataella phaffii</i> (<i>Pichia pastoris</i>): Formaldehyde Toxicity and Energy Metabolism |
| title_fullStr | Role of Dissimilative Pathway of <i>Komagataella phaffii</i> (<i>Pichia pastoris</i>): Formaldehyde Toxicity and Energy Metabolism |
| title_full_unstemmed | Role of Dissimilative Pathway of <i>Komagataella phaffii</i> (<i>Pichia pastoris</i>): Formaldehyde Toxicity and Energy Metabolism |
| title_short | Role of Dissimilative Pathway of <i>Komagataella phaffii</i> (<i>Pichia pastoris</i>): Formaldehyde Toxicity and Energy Metabolism |
| title_sort | role of dissimilative pathway of i komagataella phaffii i i pichia pastoris i formaldehyde toxicity and energy metabolism |
| topic | <i>Komagataella phaffii</i> <i>Pichia pastoris</i> methanol formaldehyde dehydrogenase dissimilative pathway |
| url | https://www.mdpi.com/2076-2607/10/7/1466 |
| work_keys_str_mv | AT julioberrios roleofdissimilativepathwayofikomagataellaphaffiiiipichiapastorisiformaldehydetoxicityandenergymetabolism AT chrispianwtheron roleofdissimilativepathwayofikomagataellaphaffiiiipichiapastorisiformaldehydetoxicityandenergymetabolism AT sebastiensteels roleofdissimilativepathwayofikomagataellaphaffiiiipichiapastorisiformaldehydetoxicityandenergymetabolism AT belenponce roleofdissimilativepathwayofikomagataellaphaffiiiipichiapastorisiformaldehydetoxicityandenergymetabolism AT edgarvelastegui roleofdissimilativepathwayofikomagataellaphaffiiiipichiapastorisiformaldehydetoxicityandenergymetabolism AT cristinabustos roleofdissimilativepathwayofikomagataellaphaffiiiipichiapastorisiformaldehydetoxicityandenergymetabolism AT claudiaaltamirano roleofdissimilativepathwayofikomagataellaphaffiiiipichiapastorisiformaldehydetoxicityandenergymetabolism AT patrickfickers roleofdissimilativepathwayofikomagataellaphaffiiiipichiapastorisiformaldehydetoxicityandenergymetabolism |