Functional characterization of a novel thermophilic exo-arabinanase from Thermothielavioides terrestris
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2020-10-01
Autores
Velasco, Josman
Oliva, Bianca
Gonçalves, Aline Larissa
Lima, Awana Silva
Ferreira, Gislene
França, Bruno Alves
Mulinari, Evandro José
Gonçalves, Thiago Augusto
Squina, Fábio Márcio
Kadowaki, Marco Antonio Seiki
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Abstract: Arabinanases from glycoside hydrolase family GH93 are enzymes with exo-activity that hydrolyze the α-1,5 bonds between arabinose residues present on arabinan. Currently, several initiatives aiming to use byproducts rich in arabinan such as pectin and sugar beet pulp as raw material to produce various compounds of interest are being developed. However, it is necessary to use robust enzymes that have an optimal performance under pH and temperature conditions used in the industrial processes. In this work, the first GH93 from the thermophilic fungus Thermothielavioides terrestris (Abn93T) was heterologously expressed in Aspergillus nidulans, purified and biochemically characterized. The enzyme is a thermophilic glycoprotein (optimum activity at 70 °C) with prolonged stability in acid pHs (4.0 to 6.5). The presence of glycosylation affected slightly the hydrolytic capacity of the enzyme, which was further increased by 34% in the presence of 1 mM CoCl2. Small-angle X-ray scattering results show that Abn93T is a globular-like-shaped protein with a slight bulge at one end. The hydrolytic mechanism of the enzyme was elucidated using capillary zone electrophoresis and molecular docking calculations. Abn93T has an ability to produce (in synergism with arabinofuranosidases) arabinose and arabinobiose from sugar beet arabinan, which can be explored as fermentable sugars and prebiotics. Key points: • Thermophilic exo-arabinanase from family GH93 • Molecular basis of arabinan depolymerization.
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Arabinan, Aspergillus nidulans, Exo-arabinanase, Sugar beet pulp, Thermothielavioides terrestris
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Applied Microbiology and Biotechnology, v. 104, n. 19, p. 8309-8326, 2020.