title: Characterization and expression of a GDSL-like lipase gene from Brassica napus in Nicotiana benthamiana.
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 GDSL-like 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) n-caprylic 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 17.7 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.
authors.affiliation: Institute of Life Sciences, Jiangsu University, Zhenjiang, 212013, People's Republic of China, xltan@ujs.edu.cn.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Tan
authors.forename: Suzhen
authors.initials: S
authors.lastname: Yan
authors.forename: Renke
authors.initials: R
authors.lastname: Tan
authors.forename: Zhiyan
authors.initials: Z
authors.lastname: Zhang
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.forename: Jie
authors.initials: J
authors.lastname: Chen
mesh_terms: D029688:Brassica napus / Q000235:genetics* / Q000378:metabolism; D004950:Esterases / Q000235:genetics* / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D008049:Lipase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D018515:Plant Leaves / Q000201:enzymology; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014026:Nicotiana / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D004950:Esterases; D008049:Lipase