Purification of industrial αamylase by reversed micellar extraction

• Published in: Biotechnology and bioengineering 1995-12, Vol.48 (6), p.745-748
• Main Authors: Chang, Qing-Long, Chen, Jia-Yong
• Format: Article
• Language: English
• Subjects: Aliquat 336; Biological and medical sciences; Biotechnology; Enzyme engineering; Fundamental and applied biological sciences. Psychology; Improved methods for extraction and purification of enzymes; Methods. Procedures. Technologies; purification; reversed micelles; α-amylase
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.260480624

The purification of industrial α‐amylase by liquid‐liquid extraction with Aliquat 336 reversed micellar solution as the extractant was studied. Seven kinds of Aliquat 336 reversed micellar solution, formed by using seven kinds of straight chain alkyl alcohols as cosolvent, have been utilized to extract industrial a‐amylase. It was found that these seven kinds of reversed micellar solution can all achieve a high protein transfer efficiency in the forward extraction process. After a full forward and backward extraction cycle, however, only the reversed micelles with n‐butanol as the cosolvent was found to be able to maintain the activity of α‐amylase in the stripping solution. By using the reversed micelles of Aliquat 336/isooctane/1% (v/v) n‐butanol to perform a full extraction cycle, it was found that 85% of the total activity of α‐amylase in the industrial a‐amylase could be recovered at the end of an extraction cycle and the specific activity of α‐amylase could be concentrated about 1.5‐fold; meanwhile, most of the neutral protease in the industrial a‐amylase could be removed. The separation factor of α‐amylase to neutral protease at the end of an extraction cycle can reach about 10. © 1995 John Wiley & Sons, Inc.