Análisis comparativo del proteoma de Aspergillus brasiliensis en cultivos sólido y líquido
The present work was focused on the characterization of Aspergillus brasiliensis proteome, assessing the culture system and sucrose concentration present in the culture medium. The work was parted into two stages. In the first stage, the effect of sucrose concentration under SSF (20 to 210 g/L, usin...
| Autor: | |
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| Tipo de recurso: | tesis doctoral |
| Estado: | Versión publicada |
| Fecha de publicación: | 2022 |
| País: | México |
| Institución: | Universidad Autónoma Metropolitana |
| Repositorio: | Repositorio Institucional de la UAM Iztapalapa |
| Idioma: | español |
| OAI Identifier: | oai:bindani.izt.uam.mx:q811kj93r |
| Acceso en línea: | https://doi.org/10.24275/uami.q811kj93r |
| Access Level: | acceso abierto |
| Palabra clave: | info:eu-repo/classification/LEM/Fermentación info:eu-repo/classification/LEM/Proteomics info:eu-repo/classification/LEM/Aspergillus info:eu-repo/classification/LEM/Fermentation info:eu-repo/classification/LEM/Proteómica info:eu-repo/classification/cti/6 |
| Sumario: | The present work was focused on the characterization of Aspergillus brasiliensis proteome, assessing the culture system and sucrose concentration present in the culture medium. The work was parted into two stages. In the first stage, the effect of sucrose concentration under SSF (20 to 210 g/L, using perlite as support) and SmF (20 to 80 g/L) over (i) extracellular inulinase and invertase production and (ii) associated variables with A. brasiliensis growth, estimated from online CO2 production measurement was evaluated. The analysis revealed a lower Lag phase in SSF compared to SmF and, decrease in the specific CO2 production rate (μCO2) in both systems as the sucrose concentration increased. The highest maximum CO2 production rate (10.45 ± 0.72 mg/h mL) and total CO2 production (138.53 ± 6.48 mg/mL) were obtained in SSF with a sucrose concentration of 210 g/L. Inulinase and invertase production in SmF was independent of the initial sucrose concentration (1.13 ± 0.02 and 5.84 ± 0.12 U/mL, respectively); meanwhile in SSF, both activities increased with the increase in the sucrose concentration, reaching maximum values of 5.88 ± 0.78 and 41.69 ± 3.60 U/mL (inulinase and invertase, respectively), at the highest sucrose concentration (210 g/L). The zymographic analysis developed with enzymatic extracts obtained with 60 g/L of sucrose under SSF and SmF showed one band of inulinase and two bands of invertases involved in sucrose hydrolysis. In the second stage, the A. brasiliensis proteomes were obtained under SSF (sucrose: 20 to 180 g/L) and SmF (sucrose: 20 to 60 g/L) conditions. The proteome was more complex in SSF than in SmF. A total of 1471 and 953 proteins were identified, respectively. In SSF, 237 proteins (88 unique, 75 abundant unregulated, and 74 abundant regulated) were studied at sucrose concentrations < 60 g/L and 343 proteins (195 unique, 98 abundant unregulated, and 50 abundant regulated) were analyzed at sucrose concentrations > 180 g/L. In SmF, 277 proteins (128 unique, 73 abundant unregulated, and 76 abundant regulated) were examined. The proteome in SSF was 3.3 (from 20 to 60 g/L) and 4.8 (from 60 to 180 g/L) times more abundant than in SmF. In SSF there was the presence of proteins involved in a greater number of processes than in SmF. The weighted spectral count of uncharacterized, transport-associated proteins, of nucleotide and amino acid metabolism was so low that it can be considered negligible in SmF. The associated carbohydrate metabolism proteins were the most abundant group in SSF (49.98 and 80.42 spectral count/mL at sucrose concentrations of 20 to 60 g/L and 60 to 180 g/L, respectively). The sum of the weighted spectral count of all the proteins involved in this metabolism was almost 12 and 19 times more abundant than in SmF, where it ranked sixth in terms of abundance. Proteins classified as miscellaneous were the second most abundant group in SSF (40.89 and 59.66 spectral count/mL with concentrations of 20 to 60 g/L and 60 to 180 g/L, respectively). This group of proteins in SmF ranked third in terms of abundance. In the proteome in SSF and SmF, proteins involved in hydrocarbon degradation, antibiotic biosynthesis, and pathogenicity were found, with a greater variety of these in CMS. In the proteome in SSF with 20 to 60 g/L of sucrose, the third most abundant group of proteins was classified as cellular component/process (40.44 spectral count/mL), while with 60 to 180 g/L of sucrose, it ranked fourth (41.85 spectral count/mL). This group of proteins in SmF was the most abundant (27.77 spectral count/mL), giving evidence that cellular processes associated with DNA replication and transcription, and cell wall synthesis predominate in this culture system.Therefore, the findings obtained in the present work show new evidence to propose that catabolism predominates over anabolism in SSF. |
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