Authors: Xiaoli Tan Suzhen Yan Renke Tan Zhiyan Zhang Zheng Wang Jie Chen
Publish Date: 2013/12/21
Volume: 33, Issue: 1, Pages: 18-23
Abstract
The GDSL esterase and lipase families play important roles in abiotic stress pathogen defense seed development and lipid metabolism Identifying the lipase activity of the putative GDSL lipase is the prerequisite for dissecting its function According to the sequence similarity and the conserved domains we cloned the Brassica napus BnGLIP gene which encodes a GDSLlike protein We failed to identify the BnGLIP lipase activity in the bacterium and yeast expression systems In this paper we expressed the BnGLIP gene by fusing a 6× His tag in Nicotiana benthamiana and purified the recombinant protein The extraction buffer contained 1 v/v ncaprylic acid and was able to remove most of the protein impurities About 50 μg of recombinant BnGLIP was obtained from 1 g of N benthamiana leaves The lipase activity was tested with the purified BnGLIP and the maximum enzyme activity reached 177 mM/mg In conclusion this study found that the recombinant protein BnGLIP expressed in tobacco system was effectively purified and was detected as a GDSL lipase
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