标题：High level expression and glycosylation of recombinant Mycobacterium tuberculosis Ala-Pro-rich antigen in Pichia pastoris
作者：Wang, Shengjun; Wang, Yaoguang; Wang, Peng George; Chen, Min; Kong, Yun
作者机构：[Wang, Shengjun; Wang, Yaoguang; Wang, Peng George; Chen, Min; Kong, Yun] Shandong Univ, Sch Life Sci, Natl Glycoengn Res Ctr, Jinan 250100, Shandong, 更多
通讯作者：Kong, Y;Kong, Y
通讯作者地址：[Kong, Y]Shandong Univ, Sch Life Sci, Natl Glycoengn Res Ctr, Jinan 250100, Shandong, Peoples R China;[Kong, Y]Shandong Univ, Shandong Prov Key Lab Ca 更多
来源：PROTEIN EXPRESSION AND PURIFICATION
关键词：Ala-Pro-rich antigen; High-level expression; Mycobacterium tuberculosis;; O-mannosylation; Pichia pastoris; Posttranslational modification
摘要：The Ala-Pro-rich Antigen (Apa) from Mycobacterium tuberculosis is a mannosylated protein with immunogenic and antigenic properties. The O-mannosylation is essential for its biological function in the process of colonization and invasion of host cells by M. tuberculosis. In this work, the gene encoding Apa was cloned from M. tuberculosis and expressed in Pichia pastoris GS115. In shake-flasks, the recombinant Apa was secreted into the culture media and purified with a yield of 0.6 g/L. Both N- and O-glycans were found in recombinant Apa. In P. pastoris the known M. tuberculosis-derived O-glycosites of Apa were modified with short chains of mannose units, and a few additional glycosylation sites were also observed. Therefore, the recombinant Apa expressed in P. pastoris has similar but not identical O-mannose patterns to the native protein from M. tuberculosis. P. pastoris and mycobacteria share similarities in the protein O-glycosylation pathway. Thus P. pastoris could be a potential powerful expression system to produce mycobacteria-derived glycoproteins.