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Partitioning and purification of extracellular β-1,3-1,4-glucanase in aqueous two-phase systems

摘要The partition behaviors of β-1,3-1,4-glucanase, α-amylase and neutral proteases from clarified and whole fermentation broths of Bacillus subtilis ZJF-1A5 were investigated. An aqueous two-phase system (polyethylene glycol (PEG)/MgSO4)was examined with regard to the effects of PEG molecular weight (MW) and concentration, MgSO4 concentration, pH and NaCl concentration on enzyme partition and extraction. The MW and concentration of PEG were found to have significant effects on enzyme partition and extraction with low MW PEG showing the greatest benefit in the partition and extraction of β-glucanase with the PEG/MgSO4 system. MgSO4 concentration influenced the partition and extraction of β-glucanase significantly. pH had little effect on β-glucanase or proteases partition but affected α-amylase partition when pH was over 7.0. The addition of NaCl had little effect on the partition behavior of β-glucanase but had very significant effects on the partitioning of α-amylase and on the neutral proteases. The partition behaviors of β-glucanase, α-amylase and proteases in whole broth were also investigated and results were similar to those obtained with clarified fermentation broth. A two-step process for purifying β-glucanase was developed, which achieved β-glucanase recovery of 65.3% and specific activity of 14027 U/mg, 6.6 times improvement over the whole broth.

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作者单位 School of Biosystem Engineering and Food Science, Zhejiang University, Hangzhou 310029, China [1]
分类号 Q814.1
发布时间 2005-10-27
基金项目
国家自然科学基金
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浙江大学学报B(英文版)

浙江大学学报B(英文版)

2005年6卷8期

825-831页

SCIMEDLINEISTICCSCDCABP

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