Production of a Fungal Punicalagin-Degrading Enzyme by Solid-State Fermentation: Studies of Purification and Characterization
The present work describes the purification of an enzyme capable of degrading punicalagin. The enzyme was produced by <i>Aspergillus niger</i> GH1 by solid-state fermentation, and the enzyme production was induced by using ellagitannins as the sole carbon source. The purification steps i...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-02-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/12/4/903 |
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| author | Pedro Aguilar-Zárate Gerardo Gutiérrez-Sánchez Mariela R. Michel Carl W. Bergmann José J. Buenrostro-Figueroa Juan A. Ascacio-Valdés Juan C. Contreras-Esquivel Cristóbal N. Aguilar |
| author_facet | Pedro Aguilar-Zárate Gerardo Gutiérrez-Sánchez Mariela R. Michel Carl W. Bergmann José J. Buenrostro-Figueroa Juan A. Ascacio-Valdés Juan C. Contreras-Esquivel Cristóbal N. Aguilar |
| author_sort | Pedro Aguilar-Zárate |
| collection | DOAJ |
| description | The present work describes the purification of an enzyme capable of degrading punicalagin. The enzyme was produced by <i>Aspergillus niger</i> GH1 by solid-state fermentation, and the enzyme production was induced by using ellagitannins as the sole carbon source. The purification steps included the concentration by lyophilization, desalting, anionic exchange, and gel filtration chromatography. The enzyme kinetic constants were calculated by using punicalagin, methyl gallate, and sugar beet arabinans. The molecular mass of the protein was estimated by SDS-PAGE. The identified bands were excised and digested using trypsin, and the peptides were submitted to HPLC-MS/MS analysis. The docking analysis was conducted, and a 3D model was created. The purification fold increases 75 times compared with the cell-free extract. The obtained <i>Km</i> values were 0.053 mM, 0.53% and 6.66 mM for punicalagin, sugar beet arabinans and methyl gallate, respectively. The optimal pH and temperature for the reaction were 5 and 40 °C, respectively. The SDS-PAGE and native PAGE analysis revealed the presence of two bands identified as α-<span style="font-variant: small-caps;">l</span>-arabinofuranosidase. Both enzymes were capable of degrading punicalagin and releasing ellagic acid. |
| first_indexed | 2024-03-11T08:49:39Z |
| format | Article |
| id | doaj.art-3205b2f67b8f40fa974827ce30f2c428 |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-11T08:49:39Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-3205b2f67b8f40fa974827ce30f2c4282023-11-16T20:32:07ZengMDPI AGFoods2304-81582023-02-0112490310.3390/foods12040903Production of a Fungal Punicalagin-Degrading Enzyme by Solid-State Fermentation: Studies of Purification and CharacterizationPedro Aguilar-Zárate0Gerardo Gutiérrez-Sánchez1Mariela R. Michel2Carl W. Bergmann3José J. Buenrostro-Figueroa4Juan A. Ascacio-Valdés5Juan C. Contreras-Esquivel6Cristóbal N. Aguilar7Engineering Department, Instituto Tecnológico de Ciudad Valles, Tecnológico Nacional de México, Ciudad Valles, San Luis Potosí C.P. 79010, MexicoComplex Carbohydrate Research Center, The University of Georgia, Athens, GA 30602, USAEngineering Department, Instituto Tecnológico de Ciudad Valles, Tecnológico Nacional de México, Ciudad Valles, San Luis Potosí C.P. 79010, MexicoComplex Carbohydrate Research Center, The University of Georgia, Athens, GA 30602, USAResearch Center in Food and Development A. C. Cd., Delicias, Chihuahua C.P. 33089, MexicoBioprocesses Research Group, Food Research Department, School of Chemistry, Universidad Autónoma de Coahuila, Saltillo, Coahuila C.P. 25280, MexicoBioprocesses Research Group, Food Research Department, School of Chemistry, Universidad Autónoma de Coahuila, Saltillo, Coahuila C.P. 25280, MexicoBioprocesses Research Group, Food Research Department, School of Chemistry, Universidad Autónoma de Coahuila, Saltillo, Coahuila C.P. 25280, MexicoThe present work describes the purification of an enzyme capable of degrading punicalagin. The enzyme was produced by <i>Aspergillus niger</i> GH1 by solid-state fermentation, and the enzyme production was induced by using ellagitannins as the sole carbon source. The purification steps included the concentration by lyophilization, desalting, anionic exchange, and gel filtration chromatography. The enzyme kinetic constants were calculated by using punicalagin, methyl gallate, and sugar beet arabinans. The molecular mass of the protein was estimated by SDS-PAGE. The identified bands were excised and digested using trypsin, and the peptides were submitted to HPLC-MS/MS analysis. The docking analysis was conducted, and a 3D model was created. The purification fold increases 75 times compared with the cell-free extract. The obtained <i>Km</i> values were 0.053 mM, 0.53% and 6.66 mM for punicalagin, sugar beet arabinans and methyl gallate, respectively. The optimal pH and temperature for the reaction were 5 and 40 °C, respectively. The SDS-PAGE and native PAGE analysis revealed the presence of two bands identified as α-<span style="font-variant: small-caps;">l</span>-arabinofuranosidase. Both enzymes were capable of degrading punicalagin and releasing ellagic acid.https://www.mdpi.com/2304-8158/12/4/903ellagitanninsellagic acid<i>Aspergillus niger</i>solid-state fermentationenzyme kinetics |
| spellingShingle | Pedro Aguilar-Zárate Gerardo Gutiérrez-Sánchez Mariela R. Michel Carl W. Bergmann José J. Buenrostro-Figueroa Juan A. Ascacio-Valdés Juan C. Contreras-Esquivel Cristóbal N. Aguilar Production of a Fungal Punicalagin-Degrading Enzyme by Solid-State Fermentation: Studies of Purification and Characterization Foods ellagitannins ellagic acid <i>Aspergillus niger</i> solid-state fermentation enzyme kinetics |
| title | Production of a Fungal Punicalagin-Degrading Enzyme by Solid-State Fermentation: Studies of Purification and Characterization |
| title_full | Production of a Fungal Punicalagin-Degrading Enzyme by Solid-State Fermentation: Studies of Purification and Characterization |
| title_fullStr | Production of a Fungal Punicalagin-Degrading Enzyme by Solid-State Fermentation: Studies of Purification and Characterization |
| title_full_unstemmed | Production of a Fungal Punicalagin-Degrading Enzyme by Solid-State Fermentation: Studies of Purification and Characterization |
| title_short | Production of a Fungal Punicalagin-Degrading Enzyme by Solid-State Fermentation: Studies of Purification and Characterization |
| title_sort | production of a fungal punicalagin degrading enzyme by solid state fermentation studies of purification and characterization |
| topic | ellagitannins ellagic acid <i>Aspergillus niger</i> solid-state fermentation enzyme kinetics |
| url | https://www.mdpi.com/2304-8158/12/4/903 |
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