Effective extraction and purification of β-xylosidase from Trichoderma koningii fermentation culture by aqueous two-phase partitioning

• Published in: Enzyme and microbial technology 2001-02, Vol.28 (2), p.196-201
• Main Authors: Pan, I-Horng, Yao, Hsin-Jan, Li, Yaw-Kuen
• Format: Article
• Language: English
• Subjects: aqueous two-phase system; Biological and medical sciences; Biotechnology; Enzyme engineering; Fundamental and applied biological sciences. Psychology; Improved methods for extraction and purification of enzymes; Methods. Procedures. Technologies; partitioning; polyethylene glycol; purification; sodium dihydrogenphosphate; sodium phosphate, monobasic; Trichoderma koningii; β-xylosidase
• ISSN: 0141-0229; 1879-0909
• DOI: 10.1016/S0141-0229(00)00291-X

Effective extraction of protein from bulk medium is an important technique in bioresearch. In the present study, we describe an extracellular β-xylosidase from the fermentation supernatant of Trichoderma koningii G-39 that was successfully extracted and purified simultaneously in a single step by using an aqueous two-phase partitioning method. This two-phase system was prepared by dissolving suitable amount of poly(ethylene glycol) (PEG) and sodium dihydrogenphosphate (NaH 2PO 4) in aqueous solution. β-Xylosidase was recovered with high yield and high concentration in the bottom salt-rich phase when 25% (w/v) PEG 1500 and 20–25% (w/v) NaH 2PO 4 were applied. Based on a 1-liter scale extraction, the purity of the enzyme was enhanced at least 33-fold. The total activity increased 422% in comparison with that in the untreated filtrate. The effectiveness and simplicity may make this technique potentially useful in various applications. The transxylosylation activity of the enzyme purified by this technique was also investigated.