Deletion of the PHO91 Gene Leads to Impaired Ability to Consume Methanol in Cells of the Methylotrophic Yeast <i>Ogataea Parapolymorpha</i>
The PHO91 and PHO87 are parts of yeast’s phosphate metabolism system. We investigated the influence of PHO91 deletion on the methanol utilization as a sole carbon source in <i>Ogataea parapolymorpha</i> through comparison of wet biomass, protein content, methanol oxidase (MOX) activity a...
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2024-05-01
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| author | Vasilina Farofonova Azamat Karginov Michael Agaphonov Tatiana Kulakovskaya |
| author_facet | Vasilina Farofonova Azamat Karginov Michael Agaphonov Tatiana Kulakovskaya |
| author_sort | Vasilina Farofonova |
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| description | The PHO91 and PHO87 are parts of yeast’s phosphate metabolism system. We investigated the influence of PHO91 deletion on the methanol utilization as a sole carbon source in <i>Ogataea parapolymorpha</i> through comparison of wet biomass, protein content, methanol oxidase (MOX) activity and acid-soluble and acid-insoluble inorganic polyphosphate (polyP) content, while growing on methanol and without any carbon source other than trace amounts from the yeast extract. The <i>Δpho91</i> strain has a lack of wet biomass, protein content and MOX activity while grown on methanol (as a <i>ΔMOX</i> strain) and has the same numbers as a WT strain in media without methanol. We found two ways of recovering methanol utilizing capabilities for <i>Δpho91</i> mutant—either adding MOX on plasmid under constitutive promoter control, or adding a <i>Δpho87</i> mutation. Both <i>Δpho91</i> + MOX and <i>Δpho91Δpho87</i> strains showed all the parameters as a WT strain on methanol as a sole carbon source and without methanol addition. Further, we compare the polyP content of the strains in methanol media, media without a sole carbon source and media with glucose. All of the WT, <i>Δpho91</i> + MOX and <i>Δpho91Δpho87</i> strains, capable of utilizing methanol, have the P<sub>i</sub>, acid-soluble and acid-insoluble polyP at the same levels with some variations. The <i>ΔMOX</i> and <i>Δpho91</i> strains, despite both not being able to utilize methanol, showed very different levels of P<sub>i</sub> and polyP’s, which can be interpreted as different mechanisms of the inner cell, leading to a loss of capability utilizing methanol in both of these strains. |
| format | Article |
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| language | English |
| publishDate | 2024-05-01 |
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| spelling | doaj-art-7c8fd1b53a994e0da4f976add29de8192025-08-20T02:43:02ZengMDPI AGProceedings2504-39002024-05-0110518310.3390/proceedings2024105083Deletion of the PHO91 Gene Leads to Impaired Ability to Consume Methanol in Cells of the Methylotrophic Yeast <i>Ogataea Parapolymorpha</i> Vasilina Farofonova0Azamat Karginov1Michael Agaphonov2Tatiana Kulakovskaya3Federal Research Center “Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences”, Institute for Biological Instrumentation of the Russian Academy of Sciences, Pushchino 142290, RussiaBach Institute of Biochemistry, Research Center of Biotechnology RAS, Moscow 119071, RussiaBach Institute of Biochemistry, Research Center of Biotechnology RAS, Moscow 119071, RussiaFederal Research Center “Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences”, Skryabin Institute of Biochemistry and Physiology of Microorganisms, Pushchino 142290, RussiaThe PHO91 and PHO87 are parts of yeast’s phosphate metabolism system. We investigated the influence of PHO91 deletion on the methanol utilization as a sole carbon source in <i>Ogataea parapolymorpha</i> through comparison of wet biomass, protein content, methanol oxidase (MOX) activity and acid-soluble and acid-insoluble inorganic polyphosphate (polyP) content, while growing on methanol and without any carbon source other than trace amounts from the yeast extract. The <i>Δpho91</i> strain has a lack of wet biomass, protein content and MOX activity while grown on methanol (as a <i>ΔMOX</i> strain) and has the same numbers as a WT strain in media without methanol. We found two ways of recovering methanol utilizing capabilities for <i>Δpho91</i> mutant—either adding MOX on plasmid under constitutive promoter control, or adding a <i>Δpho87</i> mutation. Both <i>Δpho91</i> + MOX and <i>Δpho91Δpho87</i> strains showed all the parameters as a WT strain on methanol as a sole carbon source and without methanol addition. Further, we compare the polyP content of the strains in methanol media, media without a sole carbon source and media with glucose. All of the WT, <i>Δpho91</i> + MOX and <i>Δpho91Δpho87</i> strains, capable of utilizing methanol, have the P<sub>i</sub>, acid-soluble and acid-insoluble polyP at the same levels with some variations. The <i>ΔMOX</i> and <i>Δpho91</i> strains, despite both not being able to utilize methanol, showed very different levels of P<sub>i</sub> and polyP’s, which can be interpreted as different mechanisms of the inner cell, leading to a loss of capability utilizing methanol in both of these strains.https://www.mdpi.com/2504-3900/105/1/83PHO91methanol utilizationyeasts<i>Ogataea parapolymorpha</i> |
| spellingShingle | Vasilina Farofonova Azamat Karginov Michael Agaphonov Tatiana Kulakovskaya Deletion of the PHO91 Gene Leads to Impaired Ability to Consume Methanol in Cells of the Methylotrophic Yeast <i>Ogataea Parapolymorpha</i> Proceedings PHO91 methanol utilization yeasts <i>Ogataea parapolymorpha</i> |
| title | Deletion of the PHO91 Gene Leads to Impaired Ability to Consume Methanol in Cells of the Methylotrophic Yeast <i>Ogataea Parapolymorpha</i> |
| title_full | Deletion of the PHO91 Gene Leads to Impaired Ability to Consume Methanol in Cells of the Methylotrophic Yeast <i>Ogataea Parapolymorpha</i> |
| title_fullStr | Deletion of the PHO91 Gene Leads to Impaired Ability to Consume Methanol in Cells of the Methylotrophic Yeast <i>Ogataea Parapolymorpha</i> |
| title_full_unstemmed | Deletion of the PHO91 Gene Leads to Impaired Ability to Consume Methanol in Cells of the Methylotrophic Yeast <i>Ogataea Parapolymorpha</i> |
| title_short | Deletion of the PHO91 Gene Leads to Impaired Ability to Consume Methanol in Cells of the Methylotrophic Yeast <i>Ogataea Parapolymorpha</i> |
| title_sort | deletion of the pho91 gene leads to impaired ability to consume methanol in cells of the methylotrophic yeast i ogataea parapolymorpha i |
| topic | PHO91 methanol utilization yeasts <i>Ogataea parapolymorpha</i> |
| url | https://www.mdpi.com/2504-3900/105/1/83 |
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