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Convenient one-step purification and immobilization of lipase using a genetically encoded aldehyde tag

Paper ID Volume ID Publish Year Pages File Format Full-Text
3355 165 2013 7 PDF Available
Title
Convenient one-step purification and immobilization of lipase using a genetically encoded aldehyde tag
Abstract

To avoid the unwanted and random covalent linkage between the cross-linker and enzyme's active site in covalent immobilization, a genetically encoded “aldehyde tag” was introduced into recombinant lipase and applied for the one-step purification and covalent immobilization of this enzyme. The effects of the immobilization time, temperature and the amount of enzyme were investigated, and the thermo-stability of immobilized lipase was also examined. The specific activity and the kcat/Km of the immobilized lipase using aldehyde tag (IL-AT) were 2.50 and 3.02 fold higher, respectively, than those of the traditionally immobilized lipase using glutaraldehyde (IL-GA). The newly immobilized lipase also presented better thermo-stability than the traditionally immobilized one. The results show that the recombinant enzyme could be conveniently immobilized without glutaraldehyde and that the enzyme's active site was well protected. This is a new immobilization method able to avoid glutaraldehyde or 2,4,6-trichloro-1,3,5-triazine as an activating agent. The greener method without hazardous chemicals for the one-step purification and immobilization of an enzyme using a genetically encoded “aldehyde tag” can be exploited for numerous other enzyme purification and immobilization applications.

► One-step purification and immobilization of recombinant lipase using aldehyde tag. ► The novel immobilized enzyme presented high activity, kcat/Km and thermal stability. ► Bioorthogonal covalent linkage may be appropriate protection of enzyme's active site.

Keywords
Aldehyde-tag; Bioseparations; Immobilized enyzme; Enzyme activity; Thermo-stability; Lipase
First Page Preview
Convenient one-step purification and immobilization of lipase using a genetically encoded aldehyde tag
Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 73, 15 April 2013, Pages 86–92
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering