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Bioprocess Biosyst Eng (2006) 28: 267–273
DOI 10.1007/s00449-005-0032-1
ORIGINAL PAPER
Jun-Guo Liu Æ Jian-Min Xing Æ Tian-Shi Chang
Hui-Zhou Liu
Purification of nattokinase by reverse micelles extraction
from fermentation broth: effect of temperature and phase
volume ratio
Received: 26 September 2005 / Accepted: 10 October 2005 / Published online: 8 December 2005
Springer-Verlag 2005
Abstract Nattokinase is a novel fibrinolytic enzyme that However, the presence of nutrients, amino acids, cell
is considered to be a promising agent for thrombosis debris and other contaminating proteins makes enzyme
therapy. In this study, reverse micelles extraction was recovery from whole fermentation broth a challenge.
applied to purify and concentrate nattokinase from The interaction of the surfactant with any broth con-
fermentation broth. The effects of temperature and stituents may affect the preferential separation of the
phase volume ratio used for the forward and backward desired product. Any complexation of the surfactant
extraction on the extraction process were examined. The with cell debris or broth constituents will decrease the
optimal temperature for forward and backward extrac- effective surfactant to achieve solubilization and sepa-
tion were 25C and 35C respectively. Nattokinase be- ration [2]. By now, only a few proteins, such as alkaline
came more thermosensitive during reverse micelles protease [2], a-amylase [3], inulinase [4], xylanase [5],
extraction. And it could be enriched in the stripping extracellular acid phosphatse [6], alcohol dehydrogenase
phase eight times during backward extraction. It was [7], lysozyme [8, 9] have been reported to be purified
found that nattokinase could be purified by AOT r
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