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Stability Analysis of Bacillus stearothermopilus L1 Lipase Fused with a Cellulose-binding Domain  

Hwang Sangpill (Department of Chemical Engineering, Yonsei University)
Ahn Ik-Sung (Department of Chemical Engineering, Yonsei University)
Publication Information
Biotechnology and Bioprocess Engineering:BBE / v.10, no.4, 2005 , pp. 329-333 More about this Journal
Abstract
This study was designed to investigate the stability of a lipase fused with a cellulose­binding domain (CBD) to cellulase. The fusion protein was derived from a gene cluster of a CBD fragment of a cellulase gene in Trichoderma hazianum and a lipase gene in Bacillus stearother­mophilus L1. Due to the CBD, this lipase can be immobilized to a cellulose material. Factors affecting the lipase stability were divided into the reaction-independent factors (RIF), and the re­action-dependent factors (RDF). RIF includes the reaction conditions such as pH and tempera­ture, whereas substrate limitation and product inhibition are examples of RDF. As pH 10 and $50^{\circ}C$ were found to be optimum reaction conditions for oil hydrolysis by this lipase, the stability of the free and the immobilized lipase was studied under these conditions. Avicel (microcrystal­line cellulose) was used as a support for lipase immobilization. The effects of both RIF and RDF on the enzyme activity were less for the immobilized lipase than for the free lipase. Due to the irreversible binding of CBD to Avicel and the high stability of the immobilized lipase, the enzyme activity after five times of use was over $70\%$ of the initial activity.
Keywords
stability; lipase; immobilization; cellulose-binding domain;
Citations & Related Records

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