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작성일 : 12-07-18 09:04

(2012, IF=3.045) Effects of Trx2p and Sec23p expression on stable production of hepatitis B surface antigen S domain in recombinant Saccharomyces cerevisiae.

글쓴이 : ycpark

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http://www.sciencedirect.com/science/article/pii/S0168165612002581 [1250]

Effects of Trx2p and Sec23p expression on stable production of hepatitis B surface antigen S domain in recombinant Saccharomyces cerevisiae.

Young-Kyung Park, Sang-Min Jung, Hyung-Kwon Lim, Young-Jin Son, Yong-Cheol Park* and Jin-Ho Seo
Journal of Biotechnology 160(3-4): 151–160. 2012. 08. 31.
(SCI I/F:3.045)

Abstract
The Sdomain of hepatitisB virus surfaceantigen (sHBsAg) is the primary component for vaccine development against virus infection. For stableexpression of sHBsAg in recombinantSaccharomycescerevisiae, new accessory genes necessary for foreign protein expression were screened by DNA microarray. Among 600 genes of interest, genes coding for an activating protein of ATPase in Hsp90 (Aha1p), S. cerevisiae DnaJ (Scj1p), thioredoxin 2 (Trx2p) and a GTPase-activator specific for Sar1 (Sec23p) as well as Pdi1p were selected in transcriptome analysis, which are known to facilitate disulfide bond formation or induce protein transport in the secretion pathway. Individual and combinatorial expression of SEC23, TRX2 and PDI1 increased total sHBsAg concentration by 1.9–6.5-fold, relative to the control strain expressing sHBsAg only. Additionally, moderate expression of Kex2p protease able to cut off the signal peptide enhanced the portion of the authentic sHBsAg to total sHBsAg. Fed-batch fermentation of the S. cerevisiae 2805 strain coexpressing the sHBsAg, SEC23, PDI1 and KEX2 genes resulted in 70.6 mg/L final sHBsAg concentration which was 5.6 times higher than that of the control. Transmission electron microscopic analysis of the yeast cells elucidated the effects of the accessory gene coexpression on the intracellular localization of sHBsAg. Like PDI1, coexpression of both SEC23 and/or TRX2 newly isolated in this study is expected to improve the target protein expression in S. cerevisiae.

Key words: Hepatitis B virus surface antigen; S domain, Saccharomyces cerevisiae; Accessory proteins; SEC23; TRX2

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