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首页> 外文期刊>Journal of Biotechnology >Chromatographic separation and kinetic properties of fructosyltransferase from Aureobasidium pullulans
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Chromatographic separation and kinetic properties of fructosyltransferase from Aureobasidium pullulans

机译：金黄芽孢杆菌果糖基转移酶的色谱分离和动力学性质

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摘要

Efficient chromatographic separation of fructosyltransferase from Aureobasidium pullulans was achieved on a preparative scale using a weak anion-exchanger Sepabeads FP-DA. The recovery yield was about 70% and the purification factor reached a value of 28. The molecular weight of the enzyme determined by size-exclusion chromatography was 570,000. The enzyme exhibited both hydrolytic and transferase activity when the latter was higher in the whole concentration range and completely dominating at higher sucrose concentrations. It was found that sucrose was the only donor of fructosyl moiety used in the transfer reaction. The initial rate method was used for the investigation of the kinetics of the action of fructosyltransferase on sucrose in the concentration range 30-2430mM. The initial transfructosylation rate was well fitted with a linear function of the sucrose activity where the activity coefficient was an exponentially decreasing function of the sucrose concentration.
机译：使用弱阴离子交换剂Sepabeads FP-DA，可以按制备规模从金黄色芽孢杆菌中高效分离果糖基转移酶。回收率约为70％，纯化系数达到28。通过尺寸排阻色谱法测定的酶的分子量为570,000。当后者在整个浓度范围内较高时，该酶既显示水解酶活性，又显示转移酶活性，并在较高的蔗糖浓度下完全占主导地位。发现蔗糖是转移反应中使用的果糖基部分的唯一供体。初始速率法用于研究浓度在30-2430mM范围内的果糖基转移酶对蔗糖的作用动力学。初始反果糖基化速率与蔗糖活性的线性函数很好地拟合，其中活性系数是蔗糖浓度的指数下降函数。

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