Chimeric cellobiohydrolase I expression, activity, and biochemical properties in three oleaginous yeast
Abstract Consolidated bioprocessing using oleaginous yeast is a promising modality for the economic conversion of plant biomass to fuels and chemicals. However, yeast are not known to produce effective biomass degrading enzymes naturally and this trait is essential for efficient consolidated bioproc...
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| Format: | Article |
| Language: | English |
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BMC
2021-01-01
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| Series: | Biotechnology for Biofuels |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13068-020-01856-z |
| _version_ | 1828337677162774528 |
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| author | Markus Alahuhta Qi Xu Eric P. Knoshaug Wei Wang Hui Wei Antonella Amore John O. Baker Todd Vander Wall Michael E. Himmel Min Zhang |
| author_facet | Markus Alahuhta Qi Xu Eric P. Knoshaug Wei Wang Hui Wei Antonella Amore John O. Baker Todd Vander Wall Michael E. Himmel Min Zhang |
| author_sort | Markus Alahuhta |
| collection | DOAJ |
| description | Abstract Consolidated bioprocessing using oleaginous yeast is a promising modality for the economic conversion of plant biomass to fuels and chemicals. However, yeast are not known to produce effective biomass degrading enzymes naturally and this trait is essential for efficient consolidated bioprocessing. We expressed a chimeric cellobiohydrolase I gene in three different oleaginous, industrially relevant yeast: Yarrowia lipolytica, Lipomyces starkeyi, and Saccharomyces cerevisiae to study the biochemical and catalytic properties and biomass deconstruction potential of these recombinant enzymes. Our results showed differences in glycosylation, surface charge, thermal and proteolytic stability, and efficacy of biomass digestion. L. starkeyi was shown to be an inferior active cellulase producer compared to both the Y. lipolytica and S. cerevisiae enzymes, whereas the cellulase expressed in S. cerevisiae displayed the lowest activity against dilute-acid-pretreated corn stover. Comparatively, the chimeric cellobiohydrolase I enzyme expressed in Y. lipolytica was found to have a lower extent of glycosylation, better protease stability, and higher activity against dilute-acid-pretreated corn stover. |
| first_indexed | 2024-04-13T22:18:15Z |
| format | Article |
| id | doaj.art-5ffb6d81c6ce474180ba296925f4f202 |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-04-13T22:18:15Z |
| publishDate | 2021-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-5ffb6d81c6ce474180ba296925f4f2022022-12-22T02:27:23ZengBMCBiotechnology for Biofuels1754-68342021-01-0114111110.1186/s13068-020-01856-zChimeric cellobiohydrolase I expression, activity, and biochemical properties in three oleaginous yeastMarkus Alahuhta0Qi Xu1Eric P. Knoshaug2Wei Wang3Hui Wei4Antonella Amore5John O. Baker6Todd Vander Wall7Michael E. Himmel8Min Zhang9Biosciences Center, National Renewable Energy LaboratoryBiosciences Center, National Renewable Energy LaboratoryBiosciences Center, National Renewable Energy LaboratoryBiosciences Center, National Renewable Energy LaboratoryBiosciences Center, National Renewable Energy LaboratoryBiosciences Center, National Renewable Energy LaboratoryBiosciences Center, National Renewable Energy LaboratoryBiosciences Center, National Renewable Energy LaboratoryBiosciences Center, National Renewable Energy LaboratoryBiosciences Center, National Renewable Energy LaboratoryAbstract Consolidated bioprocessing using oleaginous yeast is a promising modality for the economic conversion of plant biomass to fuels and chemicals. However, yeast are not known to produce effective biomass degrading enzymes naturally and this trait is essential for efficient consolidated bioprocessing. We expressed a chimeric cellobiohydrolase I gene in three different oleaginous, industrially relevant yeast: Yarrowia lipolytica, Lipomyces starkeyi, and Saccharomyces cerevisiae to study the biochemical and catalytic properties and biomass deconstruction potential of these recombinant enzymes. Our results showed differences in glycosylation, surface charge, thermal and proteolytic stability, and efficacy of biomass digestion. L. starkeyi was shown to be an inferior active cellulase producer compared to both the Y. lipolytica and S. cerevisiae enzymes, whereas the cellulase expressed in S. cerevisiae displayed the lowest activity against dilute-acid-pretreated corn stover. Comparatively, the chimeric cellobiohydrolase I enzyme expressed in Y. lipolytica was found to have a lower extent of glycosylation, better protease stability, and higher activity against dilute-acid-pretreated corn stover.https://doi.org/10.1186/s13068-020-01856-zMetabolic engineering: chimeric proteinOleaginous yeastCBH IConsolidated bioprocessingCellobiohydrolaseCel7A |
| spellingShingle | Markus Alahuhta Qi Xu Eric P. Knoshaug Wei Wang Hui Wei Antonella Amore John O. Baker Todd Vander Wall Michael E. Himmel Min Zhang Chimeric cellobiohydrolase I expression, activity, and biochemical properties in three oleaginous yeast Biotechnology for Biofuels Metabolic engineering: chimeric protein Oleaginous yeast CBH I Consolidated bioprocessing Cellobiohydrolase Cel7A |
| title | Chimeric cellobiohydrolase I expression, activity, and biochemical properties in three oleaginous yeast |
| title_full | Chimeric cellobiohydrolase I expression, activity, and biochemical properties in three oleaginous yeast |
| title_fullStr | Chimeric cellobiohydrolase I expression, activity, and biochemical properties in three oleaginous yeast |
| title_full_unstemmed | Chimeric cellobiohydrolase I expression, activity, and biochemical properties in three oleaginous yeast |
| title_short | Chimeric cellobiohydrolase I expression, activity, and biochemical properties in three oleaginous yeast |
| title_sort | chimeric cellobiohydrolase i expression activity and biochemical properties in three oleaginous yeast |
| topic | Metabolic engineering: chimeric protein Oleaginous yeast CBH I Consolidated bioprocessing Cellobiohydrolase Cel7A |
| url | https://doi.org/10.1186/s13068-020-01856-z |
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