极地研究 ›› 2011, Vol. 23 ›› Issue (4): 283-288.DOI: 10.3724/SP.J.1084.2011.00283

• 研究论文 • 上一篇    下一篇

低温脂肪酶基因在巴斯德毕赤酵母中的高效表达

王永杰1,2,沈继红1,2,丛柏林1,2,林学政1,2,黄晓航1,2   

  1. 1. 国家海洋局第一海洋研究所
    2. 海洋生物活性物质国家海洋局重点实验室
  • 收稿日期:2011-02-28 修回日期:2011-06-15 出版日期:2011-12-30 发布日期:2011-12-30
  • 通讯作者: 沈继红
  • 基金资助:

    863计划——极地特殊海洋微生物资源利用的关键技术研究

HIGH-LEVEL EXPRESSION OF A COLD-ADAPTIVE LIPASE GENE IN PICHIA PASTORIS

Wang Yongjie1,2Shen Jihong1,2Cong Bailin1,2Lin Xuezheng1,2Huang Xiaohang1,2   

  1. 1. First Institute of Oceanography, SOA
    2. Key Lab of Marine Bioactive Substances, SOA
  • Received:2011-02-28 Revised:2011-06-15 Online:2011-12-30 Published:2011-12-30

摘要:

通过PCR技术扩增得到了编码南极嗜冷菌低温脂肪酶948的基因序列,将其与表达载体pPIC9K连接后转化巴斯德毕赤酵母GS115,通过筛选得到了高效表达低温脂肪酶的酵母菌株,该菌株发酵48h后上清中脂肪酶活性达到27.5U/mL,普遍高于其他低温脂肪酶在毕赤酵母中的表达活性,为该低温脂肪酶的工业化应用奠定了一定的基础。

关键词: 低温脂肪酶, 表达载体pPIC9K, 巴斯德毕赤酵母, 高效表达

Abstract:

A cold-adaptive lipase gene lip-948 from Antarctic psychrophilic bacteria was ligated into the expression vector pPIC9K, which was transformed into yeast GS115 subsequently. High-level express yeast was obtained . Lipase activity of the supernant reached 27.5 U/mL after 48 h fermentation, which is higher than most of other low-temperature bacteria lipase genes expressed in Pichia pastoris.

Key words: low-temperature lipase, pPIC9K, Pichia pastoris, High-level expression