Enhancement of recombinant β-D-glucuronidase production under low-shear modeled microgravity in Pichia pastoris
Enhancement of recombinant β-D-glucuronidase production under low-shear modeled microgravity in Pichia pastoris
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DOI：
10.1002/jctb.2541
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发表时间：
2011-04-01
影响因子：
3.4
通讯作者：
Li, Chun
中科院分区：
文献类型：
--
作者：
Qi, Feng;Kaleem, Imdad;Li, Chun
背景技术背景：研究了低剪切模拟微重力（LSMMG）对真核表达宿主中重组蛋白生产的影响。该基因（GenBank Accession No.EU095019）来源于产紫青霉（Penicillium purpurogenum Stoll）（CGMCC 3. 3708）在3.2.1.31毕赤酵母GS115中表达。结果：在甲醇诱导阶段，重组毕赤酵母在LSMMG条件下的生长优于在正常重力条件下的生长。在4种不同转速下，LSMMG的PGUS表达效率比NG提高了1.51 ~ 2.21倍（P < 0.05），其中15 rpm为PGUS表达的最佳转速。结论：LSMMG环境显著提高了重组PGUS在毕赤酵母GS115中的表达和分泌。结果表明，模拟微重力技术可用于微生物宿主高效生产重组蛋白。(C)2010年化学工业协会
BACKGROUND: The effects of low-shear modeled microgravity (LSMMG) on the production of recombinant proteins in a eukaryotic expression host have been investigated. The gene (GenBank Accession No.EU095019) derived from Penicillium purpurogenum Stoll (CGMCC 3. 3708) encoding beta-D-glucuronidase (EC 3.2.1.31, PGUS) was expressed in Pichia pastoris GS115. The behavior of P. pastoris growth and recombinant PGUS production during the methanol induction stage were examined in a high-aspect ratio vessel (HARV) that could model the microgravity environment.RESULTS: The recombinant P. pastoris showed better growth under LSMMG than normal gravity (NG) during the methanol induction phase. It was also found that the efficiencies of PGUS production were enhanced 1.51 to 2.21-fold under LSMMG compared with NG control at four different rotary speeds (P < 0.05), and 15 rpm was found to be the optimal for PGUS expression. Furthermore, the efficiency of PGUS secretion was also enhanced under LSMMG (all values above 30%).CONCLUSIONS: The LSMMG environment significantly enhances production and secretion of the recombinant PGUS expressed in P. pastoris GS115. Results suggest that simulated microgravity techniques could be used for the efficient production of recombinant proteins by microbial hosts. (C) 2010 Society of Chemical Industry