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首页> 外文会议>IT in Medicine and Education (ITME), 2011 International Symposium on >Inducible expression of a β-glucuronidase from Penicillium purpurogenum in Pichia pastoris and characterization of the recombinant enzyme
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Inducible expression of a β-glucuronidase from Penicillium purpurogenum in Pichia pastoris and characterization of the recombinant enzyme

机译：紫青霉中β-葡萄糖醛酸苷酶在毕赤酵母中的诱导表达及重组酶的表征

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摘要

The present study reports the recombinant expression and partial characterization of a novel β-glucuronidase gene from Penicillium purpurogenum Li-3. It is the first time that a β-glucuronidase gene was cloned from Penicillium purpurogenum (pgus, Genbank Accession NO. EU095019). Sequence analysis indicated that pgus has 1815 base pairs, encoding 604 amino acids with the potential molecular weight of 66.7 kDa and 4 potential N-glycosylation sites. The pgus gene was successfully expressed as a functional protein (PGUS-P) in Pichia pastoris GS115. The optimal reaction temperature and pH of PGUS-P were 37.5 °C and pH5.2, respectively. While higher thermal and pH stability were observed in PGUS-P. The Km and Vmax values of PGUS-P for glycyrrhizin ammonium salt (GL) were 0.48 mM and 0.133 mM/min, respectively. The research also showed that the Mg2+, Mn2+ and Na+ have activation effect, while Ag+ and SDS has inhibition effect on the activity of catalyst. In addition, the mature PGUS-P protein exhibited a molecular mass of approximately 90 kDa on SDS-PAGE, which is much higher than its theoretical value. The results revealed that the recombinant PGUS-P may be partly N-glycosylated.
机译：本研究报道了来自青霉青霉Li-3的新型β-葡糖醛酸糖苷酶基因的重组表达和部分表征。这是首次从紫青霉菌（pgus，Genbank登录号EU095019）克隆β-葡萄糖醛酸苷酶基因。序列分析表明，pgus具有1815个碱基对，编码604个氨基酸，潜在分子量为66.7 kDa，并具有4个潜在的N-糖基化位点。 pgus基因已在巴斯德毕赤酵母GS115中成功表达为功能蛋白（PGUS-P）。 PGUS-P的最佳反应温度和pH分别为37.5°C和pH5.2。在PGUS-P中观察到较高的热稳定性和pH稳定性。甘草甜素铵盐（GL）的PGUS-P的Km和Vmax值分别为0.48 mM和0.133 mM / min。研究还表明，Mg 2 + ，Mn 2 + 和Na + 具有激活作用，而Ag +

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