Insights into the transcriptomic mechanism and characterization of endoglucanases from Aspergillus terreus in cellulose degradation.
Int J Biol Macromol
; 263(Pt 1): 130340, 2024 Apr.
Article
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| MEDLINE
| ID: mdl-38387642
ABSTRACT
Filamentous fungi are the main industrial source of cellulases which are important in the process of converting cellulose to fermentable sugars. In this study, transcriptome analysis was conducted on Aspergillus terreus NEAU-7 cultivated using corn stover and glucose as carbon sources. Four putative endoglucanases (EG5A, EG7A, EG12A, and EG12C) from A. terreus NEAU-7 were efficiently expressed in Pichia pastoris. Among them, EG7A exhibited the highest enzyme activity (75.17 U/mg) with an optimal temperature of 40 °C and pH 5.0. EG5A and EG12A displayed specific activities of 19.92 U/mg and 14.62 U/mg, respectively, at 50 °C. EG12C showed acidophilic characteristics with an optimal pH of 3.0 and a specific activity of 12.21 U/mg at 40 °C. With CMC-Na as the substrate, the Km value of EG5A, EG7A, EG12A or, EG12C was, 11.08 ± 0.87 mg/mL, 6.82 ± 0.74 mg/mL, 7.26 ± 0.64 mg/mL, and 9.88 ± 0.86 mg/mL, with Vmax values of 1258.23 ± 51.62 µmolâmin-1âmg-1, 842.65 ± 41.53 µmolâmin-1âmg-1, 499.38 ± 20.42 µmolâmin-1âmg-1, and 681.41 ± 30.08 µmolâmin-1âmg-1, respectively. The co-treatment of EG7A with the commercial cellulase increased the yield of reducing sugar by 155.77 % (filter paper) and 130.49 % (corn stover). Molecular docking assay showed the interaction energy of EG7A with cellotetraose at -10.50 kcal/mol, surpassing EG12A (-10.43 kcal/mol), EG12C (-10.28 kcal/mol), and EG5A (-9.00 kcal/mol). Root Mean Square Deviation (RMSD) and Solvent Accessible Surface Area (SASA) values revealed that the presence of cellotetraose stabilized the molecular dynamics simulation of the cellotetraose-protein complex over a 100 ns time scale. This study provides valuable insights for developing recombinant enzymes and biomass degradation technologies.
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MEDLINE
Asunto principal:
Aspergillus
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Celulasa
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En
Año:
2024
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Article