title: Prokaryotic expression, purification, and sub-cellular localization of a novel alcohol acyltransferase from apple.
abstract: The coding region of the alcohol acyltransferase gene (MdAAT2) from apple was sub-cloned into expression vectors, pET30a and pET32a, and introduced into E. coli for expression. The purified pET30a/MdAAT2 fusion proteins were used to immunize rabbits following standard protocols. The partially soluble fusion proteins had alcohol acyltransferase activity and were detected only in the pET32a/Origami B(DE3) expression system. Immunolocalization analysis indicated that MdAAT2 is mainly in the cytoplasm, in agreement with the prediction of sub-cellular localization obtained by the LOCSVMpsi program. Western blot analysis indicated that ester biosynthesis in different apple cultivars was related positively to the accumulation of MdAAT2.
authors.affiliation: College of Food Sciences, Shandong Agricultural University, Taian, 271018, P.R. China. dqli@sdau.edu.cn
authors.forename: Dapeng
authors.initials: D
authors.lastname: Li
authors.forename: Gangming
authors.initials: G
authors.lastname: Xu
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Xu
authors.forename: Tao
authors.initials: T
authors.lastname: Wu
authors.forename: Jin
authors.initials: J
authors.lastname: Shen
authors.forename: Dequan
authors.initials: D
authors.lastname: Li
authors.forename: Huairui
authors.initials: H
authors.lastname: Shu
mesh_terms: D000818:Animals; D004926:Escherichia coli / Q000502:physiology; D005638:Fruit / Q000201:enzymology* / Q000235:genetics; D027845:Malus / Q000201:enzymology* / Q000235:genetics; D015202:Protein Engineering / Q000379:methods; D011506:Proteins / Q000737:chemistry* / Q000145:classification / Q000276:immunology / Q000378:metabolism*; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D013347:Subcellular Fractions / Q000201:enzymology*
chemical_list: D011506:Proteins; D011994:Recombinant Proteins; C079238:alcohol O-acetyltransferase