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Characterization of a recombinant xylose tolerant β-xylosidase from Humicola grisea var. thermoidea and its use in sugarcane bagasse hydrolysis
- Source :
- International journal of biological macromolecules. 105(Pt 1)
- Publication Year :
- 2016
-
Abstract
- One full-length β-xylosidase gene (hxylA) was identified from the Humicola grisea var. thermoidea genome and the cDNA was successfully expressed by Pichia pastoris SMD1168. An optimization of enzyme production was carried out, and methanol was found to be the most important parameter. The purified enzyme was characterized and showed the optimal conditions for the highest activity at pH 7.0 and 50°C, being thermostable by maintaining 41% of its activity after 12h incubated at 50°C. HXYLA is a bifunctional enzyme; it showed both β-xylosidase and α-arabinfuranosidase activities. The Km and Vmax values were 1.3mM and 39.1U/mg, respectively, against 4-nitrophenyl β-xylopyranoside. HXYLA showed a relatively strong tolerance to xylose with high Ki value of 603mM, with the xylose being a non-competitive inhibitor. HXYLA was successfully used simultaneously and sequentially with an endo-xylanase for analysis of synergism in the degradation of commercial xylans. Furthermore, commercial cellulases supplementation with HXYLA during sugarcane bagasse hydrolysis increased hydrolysis in 29%. HXYLA is distinguished from other β-xylosidases by the attractive characteristics for industrial applications such as thermostability, high tolerance xylose and saccharification of biomass by convert xylan into fementable monosaccharides and improve cellulose hydrolysis.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Cellulase
Xylose
01 natural sciences
Biochemistry
Pichia
Pichia pastoris
Substrate Specificity
03 medical and health sciences
chemistry.chemical_compound
Hydrolysis
Structural Biology
010608 biotechnology
Cellulose
Molecular Biology
Thermostability
biology
General Medicine
Hydrogen-Ion Concentration
biology.organism_classification
Xylan
Recombinant Proteins
Saccharum
030104 developmental biology
Xylosidases
chemistry
Metals
biology.protein
Bagasse
Sequence Analysis
Subjects
Details
- ISSN :
- 18790003
- Volume :
- 105
- Issue :
- Pt 1
- Database :
- OpenAIRE
- Journal :
- International journal of biological macromolecules
- Accession number :
- edsair.doi.dedup.....1f44d9af0d07d70b74200ec63ee88681