Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates
Background: Xylanases and β-d-xylosidases are the most important enzymes responsible for the degradation of xylan, the second main constituent of plant cell walls. Results: In this study, the main extracellular xylanase (XYL I) and β-xylosidase (BXYL I) from the fungus Penicillium janczewskii were p...
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doaj-2b04916df44047d88c355e5c639090652020-11-25T02:25:40ZengElsevierElectronic Journal of Biotechnology0717-34582016-09-0123C546210.1016/j.ejbt.2016.08.001Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substratesCésar Rafael Fanchini Terrasan0José Manuel Guisan1Eleonora Cano Carmona2Biochemistry and Microbiology Department, Biosciences Institute, Univ Estadual Paulista – UNESP, PO Box 199, Av. 24 A, no. 1515, Bela Vista, 13506-900 Rio Claro, SP, BrazilDepartamento de Biocatálisis, Instituto de Catálisis, CSIC (Consejo Superior de Investigaciones Científicas), Campus Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, SpainBiochemistry and Microbiology Department, Biosciences Institute, Univ Estadual Paulista – UNESP, PO Box 199, Av. 24 A, no. 1515, Bela Vista, 13506-900 Rio Claro, SP, BrazilBackground: Xylanases and β-d-xylosidases are the most important enzymes responsible for the degradation of xylan, the second main constituent of plant cell walls. Results: In this study, the main extracellular xylanase (XYL I) and β-xylosidase (BXYL I) from the fungus Penicillium janczewskii were purified, characterized and applied for the hydrolysis of different substrates. Their molecular weights under denaturing and non-denaturing conditions were, respectively, 30.4 and 23.6 kDa for XYL I, and 100 and 200 kDa for BXYL I, indicating that the latter is homodimeric. XYL I is highly glycosylated (78%) with optimal activity in pH 6.0 at 65°C, while BXYL I presented lower sugar content (10.5%) and optimal activity in pH 5.0 at 75°C. The half-lives of XYL I at 55, 60 and 65°C were 125, 16 and 6 min, respectively. At 60°C, BXYL I retained almost 100% of the activity after 6 h. NH4+, Na+, DTT and β-mercaptoethanol stimulated XYL I, while activation of BXYL I was not observed. Interestingly, XYL I was only partially inhibited by Hg2+, while BXYL I was completely inhibited. Xylobiose, xylotriose and larger xylooligosaccharides were the main products from xylan hydrolysis by XYL I. BXYL I hydrolyzed xylobiose and larger xylooligosaccharides with no activity against xylans. Conclusion: The enzymes act synergistically in the degradation of xylans, and present industrial characteristics especially in relation to optimal activity at high temperatures, prolonged stability of BXYL I at 60°C, and stability of XYL I in wide pH range.http://www.sciencedirect.com/science/article/pii/S0717345816300744Xylanolytic enzymesEnzyme characterizationEnzyme purificationXylan hydrolysisXylooligosaccharides hydrolysis |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
César Rafael Fanchini Terrasan José Manuel Guisan Eleonora Cano Carmona |
spellingShingle |
César Rafael Fanchini Terrasan José Manuel Guisan Eleonora Cano Carmona Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates Electronic Journal of Biotechnology Xylanolytic enzymes Enzyme characterization Enzyme purification Xylan hydrolysis Xylooligosaccharides hydrolysis |
author_facet |
César Rafael Fanchini Terrasan José Manuel Guisan Eleonora Cano Carmona |
author_sort |
César Rafael Fanchini Terrasan |
title |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates |
title_short |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates |
title_full |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates |
title_fullStr |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates |
title_full_unstemmed |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates |
title_sort |
xylanase and β-xylosidase from penicillium janczewskii: purification, characterization and hydrolysis of substrates |
publisher |
Elsevier |
series |
Electronic Journal of Biotechnology |
issn |
0717-3458 |
publishDate |
2016-09-01 |
description |
Background: Xylanases and β-d-xylosidases are the most important enzymes responsible for the degradation of xylan, the second main constituent of plant cell walls.
Results: In this study, the main extracellular xylanase (XYL I) and β-xylosidase (BXYL I) from the fungus Penicillium janczewskii were purified, characterized and applied for the hydrolysis of different substrates. Their molecular weights under denaturing and non-denaturing conditions were, respectively, 30.4 and 23.6 kDa for XYL I, and 100 and 200 kDa for BXYL I, indicating that the latter is homodimeric. XYL I is highly glycosylated (78%) with optimal activity in pH 6.0 at 65°C, while BXYL I presented lower sugar content (10.5%) and optimal activity in pH 5.0 at 75°C. The half-lives of XYL I at 55, 60 and 65°C were 125, 16 and 6 min, respectively. At 60°C, BXYL I retained almost 100% of the activity after 6 h. NH4+, Na+, DTT and β-mercaptoethanol stimulated XYL I, while activation of BXYL I was not observed. Interestingly, XYL I was only partially inhibited by Hg2+, while BXYL I was completely inhibited. Xylobiose, xylotriose and larger xylooligosaccharides were the main products from xylan hydrolysis by XYL I. BXYL I hydrolyzed xylobiose and larger xylooligosaccharides with no activity against xylans.
Conclusion: The enzymes act synergistically in the degradation of xylans, and present industrial characteristics especially in relation to optimal activity at high temperatures, prolonged stability of BXYL I at 60°C, and stability of XYL I in wide pH range. |
topic |
Xylanolytic enzymes Enzyme characterization Enzyme purification Xylan hydrolysis Xylooligosaccharides hydrolysis |
url |
http://www.sciencedirect.com/science/article/pii/S0717345816300744 |
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