Preparation of cross-linked tyrosinase aggregates

被引:138
作者
Aytar, Burcu Selin [1 ]
Bakir, Ufuk [1 ]
机构
[1] Middle E Tech Univ, Dept Chem Engn, TR-06531 Ankara, Turkey
关键词
immobilization; mushroom tyrosinase; cross-linked enzyme aggregates; glutaraldehyde; ammonium sulfate;
D O I
10.1016/j.procbio.2007.11.001
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Tyrosinase from mushroom was immobilized as a cross-linked enzyme aggregate (CLEA) via precipitation with ammonium sulfate and cross-linking with glutaraldehyde. The effects of precipitation and cross-linking on CLEA activity were investigated and the immobilized tyrosinase was characterized. Sixty percent ammonium sulfate saturation and 2% glutaraldehyde were used; a 3-h cross-linking reaction at room temperature, at pH 7.0 was performed; particle sizes of the aggregates were reduced; consequently, 100% activity recovery was achieved in CLEAs with enhanced thermal and storage stabilities. Slight changes in optimum pH and temperature values of the enzyme were recorded after immobilization. Although immobilization did not affect V-max, substrate affinity of the enzyme increased. Highly stable CLEAs were also prepared from crude mushroom tyrosinase with 100% activity recovery. (c) 2007 Elsevier Ltd. All rights reserved.
引用
收藏
页码:125 / 131
页数:7
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