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 ext...
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Universidade Federal de Uberlandia
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my.utm.797012019-01-28T06:38:12Z http://eprints.utm.my/id/eprint/79701/ Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum Khalili, E. Huyop, F. Javed, M. A. Mahat, N. A. Batumalaie, K. Abdul Wahab, R. QD Chemistry 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. Universidade Federal de Uberlandia 2018 Article PeerReviewed application/pdf en http://eprints.utm.my/id/eprint/79701/1/RoswaniraAbdulWahab2018_AssessmentsontheCatalyticandKinetic.pdf Khalili, E. and Huyop, F. and Javed, M. A. and Mahat, N. A. and Batumalaie, K. and Abdul Wahab, R. (2018) Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum. Bioscience Journal, 34 (4). pp. 830-847. ISSN 1516-3725 http://dx.doi.org/10.14393/BJ-v34n1a2018-39384 DOI:10.14393/BJ-v34n1a2018-39384 |
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QD Chemistry Khalili, E. Huyop, F. Javed, M. A. Mahat, N. A. Batumalaie, K. Abdul Wahab, R. Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum |
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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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Article |
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Khalili, E. Huyop, F. Javed, M. A. Mahat, N. A. Batumalaie, K. Abdul Wahab, R. |
author_facet |
Khalili, E. Huyop, F. Javed, M. A. Mahat, N. A. Batumalaie, K. Abdul Wahab, R. |
author_sort |
Khalili, E. |
title |
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_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_sort |
assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus trichoderma harzianum |
publisher |
Universidade Federal de Uberlandia |
publishDate |
2018 |
url |
http://eprints.utm.my/id/eprint/79701/1/RoswaniraAbdulWahab2018_AssessmentsontheCatalyticandKinetic.pdf http://eprints.utm.my/id/eprint/79701/ http://dx.doi.org/10.14393/BJ-v34n1a2018-39384 |
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