Cloning, Purification, and Characterization of Recombinant Thermostable β-Xylanase Tnap-0700 from Thermotoga naphthophila
Journal Article
·
· Applied Biochemistry and Biotechnology
The gene of a β-xylanase (Tnap-0700) was cloned from a hyperthermophilic Thermotoga naphthophila strain ATCC BAA-489 and expressed in Escherichia coli BL21 (DE3) via pET-21a (+) as an expression vector. The growth steps were upgraded for highest β-xylanase expression via several factors, for example, IPTG concentration, time of induction, pH, and temperature. The pH and temperature optima for the extreme expression of β-xylanase were 7.0 pH and 37 °C, correspondingly. Recombinant enzyme purified by heat treatment process, then later by immobilized metal ion affinity chromatography. Molecular mass of the purified β-xylanase was 38 kDa observed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was stable at room temperature for 30 days. It exhibited high stability over wide series of temperature 50–90 °C and pH 4.0–9.0 upon the addition of 1 mM Ca{sup +2} and reduced in the existence of Cu{sup +2} and EDTA. The addition of about 10–30% different organic solvents have no considerable effect on enzyme. However, SDSF and urea acting as an inhibitor leads to decrease in the enzyme activity. The β-xylanase enzyme was active to hydrolyze xylan from beechwood forming xylose. Thermostable β-xylanase causes the breakdown of complex carbohydrates into monosaccharide components. This thermostable β-xylanase revealed remarkable properties, which make it an encouraging candidate for various industrial applications especially in the alteration of renewable biomaterials into ethanol production, and biofuels from lignocellulosics has acknowledged much devotion subsequently in the last decade.
- OSTI ID:
- 22927455
- Journal Information:
- Applied Biochemistry and Biotechnology, Journal Name: Applied Biochemistry and Biotechnology Journal Issue: 4 Vol. 189; ISSN ABIBDL; ISSN 0273-2289
- Country of Publication:
- United States
- Language:
- English
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