Production, purification, and characterization of a thermally stable, Acidophilic Cellulase from Aspergillus awamori AFE1 isolated from Longhorn beetle (Cerambycidae latreille)
Abstract Background The relentless pursuit of cost-effective cellulase, a key enzyme in the biocatalytic conversion of cellulose, has led to exploring insect guts as potential sources of biocatalysts. Herbivorous insects’ intestinal tracts are recognized as rich reservoirs of cellulolytic microorgan...
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BMC
2025-06-01
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| Series: | Microbial Cell Factories |
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| Online Access: | https://doi.org/10.1186/s12934-025-02755-4 |
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| author | Ayoola E. Afe Olusola T. Lawal Olufemi S. Bamidele Farhad Badshah Bukola R. Oyelere Andrew N. Efomah Mostafa A. Abdel-Maksoud Sabiha Fatima Abdulaziz Alamri Mohamed A. El-Tayeb David M. Sanni |
| author_facet | Ayoola E. Afe Olusola T. Lawal Olufemi S. Bamidele Farhad Badshah Bukola R. Oyelere Andrew N. Efomah Mostafa A. Abdel-Maksoud Sabiha Fatima Abdulaziz Alamri Mohamed A. El-Tayeb David M. Sanni |
| author_sort | Ayoola E. Afe |
| collection | DOAJ |
| description | Abstract Background The relentless pursuit of cost-effective cellulase, a key enzyme in the biocatalytic conversion of cellulose, has led to exploring insect guts as potential sources of biocatalysts. Herbivorous insects’ intestinal tracts are recognized as rich reservoirs of cellulolytic microorganisms. This study investigates cellulase production, purification, and characterization from Aspergillus awamori AFE1, isolated from the gut of longhorn beetles (Cerambycidae latreille). Basic cellulase production parameters were optimized. The cellulase produced under optimum conditions was purified by ammonium sulphate precipitation and chromatographic methods, followed by characterization of the purified enzyme. Results Optimum cellulase production was observed at pH 5 and 30 °C, using cellulose and NaNO3 as carbon and nitrogen sources. Cellulase was purified to homogeneity, with a molecular weight of 48.5 kDa. The cellulase exhibited optimal activity at pH 5.0 and maintained stability at an acidic pH of 4.0, showing 80% activity after 2 h and 40% activity remaining after 6 h. The optimal temperature for cellulase activity was 60 °C, with maximal stability at 30 °C, retaining 63% of its initial activity after 2 h. However, significant activity of 50% was noted at 50 °C for 2 h. Interestingly, the enzyme showed great stability against organic solvents up to 4 h and retained significant enzymatic activity after 5 h. Cellulase activity was also enhanced by divalent metal ions, Fe2+ and Zn2+, but was markedly inhibited by urea and EDTA, and monovalent Na+, K+, including some divalent metal ions, Cu2+ and Mn2+. It displayed Km and Vmax values of 3.86 mM and 0.3159 mg/mL/min, respectively. Conclusion This study has shown Aspergillus awamori AFE1, isolated from the Longhorn beetle gut, as a unique source of acid-stable, thermostable, and organic solvent-resistant cellulase with industrial potential. Its unique enzymatic properties offer promising applications in biofuel production and lignocellulosic biomass conversion. |
| format | Article |
| id | doaj-art-e71aeb0b6b73408d95ce9bb53ac5d6b8 |
| institution | Matheson Library |
| issn | 1475-2859 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | BMC |
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| series | Microbial Cell Factories |
| spelling | doaj-art-e71aeb0b6b73408d95ce9bb53ac5d6b82025-06-29T11:19:12ZengBMCMicrobial Cell Factories1475-28592025-06-0124111510.1186/s12934-025-02755-4Production, purification, and characterization of a thermally stable, Acidophilic Cellulase from Aspergillus awamori AFE1 isolated from Longhorn beetle (Cerambycidae latreille)Ayoola E. Afe0Olusola T. Lawal1Olufemi S. Bamidele2Farhad Badshah3Bukola R. Oyelere4Andrew N. Efomah5Mostafa A. Abdel-Maksoud6Sabiha Fatima7Abdulaziz Alamri8Mohamed A. El-Tayeb9David M. Sanni10Enzyme and Microbial Technology Unit, Department of Biochemistry, School of Life Sciences, Federal University of TechnologyEnzyme and Microbial Technology Unit, Department of Biochemistry, School of Life Sciences, Federal University of TechnologyEnzyme and Microbial Technology Unit, Department of Biochemistry, School of Life Sciences, Federal University of TechnologyShenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Key Laboratory of Livestock and Poultry Multi-Omics of MARA, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural SciencesDepartment of Microbiology, Federal University of TechnologySchool of Environmental Science and Engineering/Tianjin Key Lab of Biomass-Wastes Utilization/Key Laboratory of Ministry of Education, Tianjin UniversityBotany and Microbiology Department, College of Science, King Saud UniversityDepartment of Clinical Laboratory Science, College of Applied Medical Sciences, King Saud UniversityBiochemistry Department, College of Science, King Saud UniversityBotany and Microbiology Department, College of Science, King Saud UniversityEnzyme and Microbial Technology Unit, Department of Biochemistry, School of Life Sciences, Federal University of TechnologyAbstract Background The relentless pursuit of cost-effective cellulase, a key enzyme in the biocatalytic conversion of cellulose, has led to exploring insect guts as potential sources of biocatalysts. Herbivorous insects’ intestinal tracts are recognized as rich reservoirs of cellulolytic microorganisms. This study investigates cellulase production, purification, and characterization from Aspergillus awamori AFE1, isolated from the gut of longhorn beetles (Cerambycidae latreille). Basic cellulase production parameters were optimized. The cellulase produced under optimum conditions was purified by ammonium sulphate precipitation and chromatographic methods, followed by characterization of the purified enzyme. Results Optimum cellulase production was observed at pH 5 and 30 °C, using cellulose and NaNO3 as carbon and nitrogen sources. Cellulase was purified to homogeneity, with a molecular weight of 48.5 kDa. The cellulase exhibited optimal activity at pH 5.0 and maintained stability at an acidic pH of 4.0, showing 80% activity after 2 h and 40% activity remaining after 6 h. The optimal temperature for cellulase activity was 60 °C, with maximal stability at 30 °C, retaining 63% of its initial activity after 2 h. However, significant activity of 50% was noted at 50 °C for 2 h. Interestingly, the enzyme showed great stability against organic solvents up to 4 h and retained significant enzymatic activity after 5 h. Cellulase activity was also enhanced by divalent metal ions, Fe2+ and Zn2+, but was markedly inhibited by urea and EDTA, and monovalent Na+, K+, including some divalent metal ions, Cu2+ and Mn2+. It displayed Km and Vmax values of 3.86 mM and 0.3159 mg/mL/min, respectively. Conclusion This study has shown Aspergillus awamori AFE1, isolated from the Longhorn beetle gut, as a unique source of acid-stable, thermostable, and organic solvent-resistant cellulase with industrial potential. Its unique enzymatic properties offer promising applications in biofuel production and lignocellulosic biomass conversion.https://doi.org/10.1186/s12934-025-02755-4Longhorned beetleAspergillus AwamoriCellulaseEnzyme purification |
| spellingShingle | Ayoola E. Afe Olusola T. Lawal Olufemi S. Bamidele Farhad Badshah Bukola R. Oyelere Andrew N. Efomah Mostafa A. Abdel-Maksoud Sabiha Fatima Abdulaziz Alamri Mohamed A. El-Tayeb David M. Sanni Production, purification, and characterization of a thermally stable, Acidophilic Cellulase from Aspergillus awamori AFE1 isolated from Longhorn beetle (Cerambycidae latreille) Microbial Cell Factories Longhorned beetle Aspergillus Awamori Cellulase Enzyme purification |
| title | Production, purification, and characterization of a thermally stable, Acidophilic Cellulase from Aspergillus awamori AFE1 isolated from Longhorn beetle (Cerambycidae latreille) |
| title_full | Production, purification, and characterization of a thermally stable, Acidophilic Cellulase from Aspergillus awamori AFE1 isolated from Longhorn beetle (Cerambycidae latreille) |
| title_fullStr | Production, purification, and characterization of a thermally stable, Acidophilic Cellulase from Aspergillus awamori AFE1 isolated from Longhorn beetle (Cerambycidae latreille) |
| title_full_unstemmed | Production, purification, and characterization of a thermally stable, Acidophilic Cellulase from Aspergillus awamori AFE1 isolated from Longhorn beetle (Cerambycidae latreille) |
| title_short | Production, purification, and characterization of a thermally stable, Acidophilic Cellulase from Aspergillus awamori AFE1 isolated from Longhorn beetle (Cerambycidae latreille) |
| title_sort | production purification and characterization of a thermally stable acidophilic cellulase from aspergillus awamori afe1 isolated from longhorn beetle cerambycidae latreille |
| topic | Longhorned beetle Aspergillus Awamori Cellulase Enzyme purification |
| url | https://doi.org/10.1186/s12934-025-02755-4 |
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