Liquid-liquid extraction of an extracellular alkaline protease from fermentation broth using aqueous two-phase and reversed micelles systems

• Published in: World journal of microbiology & biotechnology Vol. 21; no. 5; pp. 655 - 659
• Main Authors: PORTO, T. S, MONTEIRO, T. I. R, MOREIRA, K. A, LIMA-FILHO, J. L, SILVA, M. P. C, PORTO, A. L. F, CARNEIRO-DA-CUNHA, M. G
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01-07-2005; Springer Nature B.V
• Subjects: Biological and medical sciences; Biotechnology; Enzyme engineering; Fermentation; Fundamental and applied biological sciences. Psychology; Improved methods for extraction and purification of enzymes; Methods. Procedures. Technologies; Polyethylene glycol; Potassium; Proteases; Studies; Alkaline proteinase; Enzyme; Liquid liquid extraction; liquid-liquid extraction; proteases; Fermentation; Extracellular; Peptidases; Biphasic system; Hydrolases; reversed micelles; Aqueous two-phase systems; Reverse micelle
• ISSN: 0959-3993; 1573-0972
• DOI: 10.1007/s11274-004-3570-9

The study of recovery of an extracellular alkaline protease from fermentation broth produced by Norcadiopsis sp, was carried out with liquid-liquid extraction through sodium di-(2-ethylhexyl) sulphosuccinate/isooctane reversed micelles systems and aqueous two-phase systems (polyethylene glycol/potassium phosphate). The best conditions for extraction and back-extraction with the reversed micelles system was obtained at pH 9.0 and pH 5.0, respectively, showing a yield of protein of 6.16%, a specific activity of 4.10U/ml and a purification factor of 1.80. The studies using aqueous two-phase systems of polyethylene glycol/potassium phosphate at pH 10.0 showed purification factors of 2 and 5, and protein yield of 11 and 4%, respectively, for polyethylene glycol 550/potassium phosphate and polyethylene glycol 8000/potassium phosphate. The results indicate that the aqueous two-phase systems are more attractive as a first step in the isolation and purification processes.[PUBLICATION ABSTRACT]