Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum
Due to the toxicity and inefficiency of chemical fungicides to control infestation of Macrophomina phaseolina (Tassi) Goid which causes charcoal rot in plants, a biotechnological approach using β-glucosidase (EC.3.2.1) as the alternative bioactive ingredient in fungicide is hereby, proposed. The e...
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| Format: | Article |
| Language: | English |
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Universidade Federal de Uberlândia
2018-08-01
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| Series: | Bioscience Journal |
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| Online Access: | https://seer-dev.ufu.br/index.php/biosciencejournal/article/view/39384 |
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| author | Elham Khalili Fahrul Huyop Muhammad Arshad Javed Naji Arafat Mahat Kalaivani Batumalaie Roswanira Abdul Wahab |
| author_facet | Elham Khalili Fahrul Huyop Muhammad Arshad Javed Naji Arafat Mahat Kalaivani Batumalaie Roswanira Abdul Wahab |
| author_sort | Elham Khalili |
| collection | DOAJ |
| description |
Due to the toxicity and inefficiency of chemical fungicides to control infestation of Macrophomina phaseolina (Tassi) Goid which causes charcoal rot in plants, a biotechnological approach using β-glucosidase (EC.3.2.1) as the alternative bioactive ingredient in fungicide is hereby, proposed. The extracellular enzyme was isolated from a highly efficient fungal antagonist, Trichoderma harzianum T12. The highly similar molecular masses obtained using SDS-PAGE (96 kDa) and MALDI-TOF mass spectrometry (98.3 kDa) affirmed that the β-glucosidase was purified to homogeneity. Consequently, optimum catalytic parameters that rendered the highest enzyme activity were found to be: 45˚C, pH 7, inoculum size of 10 % (w/v), supplementation with metal ions Zn2+ and Mn2+ ions, and Tween 80. Addition of wheat bran and (NH4)2SO4 as carbon and nitrogen sources also improved enzyme activity. BLASTn showed the sequence of β-glucosidase T12 was highly identical to other β-glucosidases viz. T. harzianum strain IOC-3844 (99%), T. gamsii and T. virens bgl1 (86 %) as well as T. reesei strain SJVTR and T. viride strain AS 3.3711 (84 %). Kinetic assessment showed that β-glucosidase T12 catalyzes hydrolytic activity is characterized by a Km of 0.79 mM and Vmax of 8.45 mM min-1 mg-1 protein, with a corresponding kcat of 10.69 s-1.
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| first_indexed | 2024-12-17T09:21:07Z |
| format | Article |
| id | doaj.art-ac434832f8794ce6b79bcf65bb57a9d1 |
| institution | Directory Open Access Journal |
| issn | 1981-3163 |
| language | English |
| last_indexed | 2024-12-17T09:21:07Z |
| publishDate | 2018-08-01 |
| publisher | Universidade Federal de Uberlândia |
| record_format | Article |
| series | Bioscience Journal |
| spelling | doaj.art-ac434832f8794ce6b79bcf65bb57a9d12022-12-21T21:54:45ZengUniversidade Federal de UberlândiaBioscience Journal1981-31632018-08-01344Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum Elham Khalili0Fahrul HuyopMuhammad Arshad JavedNaji Arafat MahatKalaivani BatumalaieRoswanira Abdul WahabDept. of Chemistry, Faculty of Science, Universiti Teknologi Malaysia, 81310 Skudai, Johor, Malaysia Due to the toxicity and inefficiency of chemical fungicides to control infestation of Macrophomina phaseolina (Tassi) Goid which causes charcoal rot in plants, a biotechnological approach using β-glucosidase (EC.3.2.1) as the alternative bioactive ingredient in fungicide is hereby, proposed. The extracellular enzyme was isolated from a highly efficient fungal antagonist, Trichoderma harzianum T12. The highly similar molecular masses obtained using SDS-PAGE (96 kDa) and MALDI-TOF mass spectrometry (98.3 kDa) affirmed that the β-glucosidase was purified to homogeneity. Consequently, optimum catalytic parameters that rendered the highest enzyme activity were found to be: 45ËšC, pH 7, inoculum size of 10 % (w/v), supplementation with metal ions Zn2+ and Mn2+ ions, and Tween 80. Addition of wheat bran and (NH4)2SO4 as carbon and nitrogen sources also improved enzyme activity. BLASTn showed the sequence of β-glucosidase T12 was highly identical to other β-glucosidases viz. T. harzianum strain IOC-3844 (99%), T. gamsii and T. virens bgl1 (86 %) as well as T. reesei strain SJVTR and T. viride strain AS 3.3711 (84 %). Kinetic assessment showed that β-glucosidase T12 catalyzes hydrolytic activity is characterized by a Km of 0.79 mM and Vmax of 8.45 mM min-1 mg-1 protein, with a corresponding kcat of 10.69 s-1. https://seer-dev.ufu.br/index.php/biosciencejournal/article/view/39384β-glucosidaseTrichoderma harzianumKinetic assessmentMacrophomina phaseolina |
| spellingShingle | Elham Khalili Fahrul Huyop Muhammad Arshad Javed Naji Arafat Mahat Kalaivani Batumalaie Roswanira Abdul Wahab Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum Bioscience Journal β-glucosidase Trichoderma harzianum Kinetic assessment Macrophomina phaseolina |
| title | Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum |
| title_full | Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum |
| title_fullStr | Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum |
| title_full_unstemmed | Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum |
| title_short | Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum |
| title_sort | assessments on the catalytic and kinetic properties of beta glucosidase isolated from a highly efficient antagonistic fungus trichoderma harzianum |
| topic | β-glucosidase Trichoderma harzianum Kinetic assessment Macrophomina phaseolina |
| url | https://seer-dev.ufu.br/index.php/biosciencejournal/article/view/39384 |
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