• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Bioprocess and Biosystems Engineering >Biochemical characterization and high-level production of oxidized polyvinyl alcohol hydrolase from Sphingopyxis sp. 113P3 expressed in methylotrophic Pichia pastoris
【24h】

Biochemical characterization and high-level production of oxidized polyvinyl alcohol hydrolase from Sphingopyxis sp. 113P3 expressed in methylotrophic Pichia pastoris

机译：Sphingopyxis sp。的氧化聚乙烯醇水解酶的生化特性和高产。在甲基营养毕赤酵母中表达的113P3

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The Sphingopyxis sp. 113P3 gene oph, encoding oxidized polyvinyl alcohol hydrolase (OPH), was optimized with the preferred codons of Pichia pastoris and ligated into the pPIC9K vector behind the α-factor signal sequence. The vector was then transfected into P. pastoris GS115 and genomic integration was confirmed. Large-scale production of recombinant protein was performed by induction with 14.4 g/L methanol at 22 ℃ in a 3-L bio-reactor. The maximal OPH activity obtained was 68.4 U/mL, which is the highest activity reported. The optimal pH and temperature of recombinant OPH were 8.0 and 45 ℃, respectively. OPH activity was stable over a pH range of 5.0-8.5, and at a maximal temperature of 45 ℃. The K_(cat)/K_m of recombinant OPH was 598 mM~(-1) s~(-1) which was 4.27-fold higher than that of recombinant OPH derived from Escherichia coli. The improved catalytic efficiency of OPH expressed in recombinant P. pastoris makes it favorable for industrial applications.
机译：Sphingopyxis sp。用巴斯德毕赤酵母的优选密码子优化了编码氧化聚乙烯醇水解酶（OPH）的113P3基因oph，并将其连接到α因子信号序列后面的pPIC9K载体中。然后将该载体转染到巴斯德毕赤酵母GS115中，并证实了基因组整合。通过在3升生物反应器中于22℃下用14.4克/升甲醇进行诱导，可以大规模生产重组蛋白。获得的最大OPH活性为68.4 U / mL，是报道的最高活性。重组OPH的最适pH和温度分别为8.0和45℃。 OPH活性在5.0-8.5的pH范围内稳定，最高温度为45℃。重组OPH的K_（cat）/ K_m为598 mM〜（-1）s〜（-1），比大肠杆菌来源的重组OPH高4.27倍。重组巴斯德毕赤酵母中表达的OPH的催化效率提高，使其适合工业应用。

著录项

  • 来源
    《Bioprocess and Biosystems Engineering》 |2014年第5期|777-782|共6页
  • 作者

    Yu Yang; Long Liu; Jianghua Li; Guocheng Du; Jian Chen;

  • 作者单位

    Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China,Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;

    Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China,Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;

    Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China,Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;

    Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China,Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;

    National Engineering of Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China;

  • 收录信息
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Pichia pastoris; Polyvinyl alcohol; Oxidized PVA hydrolase; Sphingopyxis sp. 113P3;

    机译：巴斯德毕赤酵母;聚乙烯醇;氧化的PVA水解酶;鞘氨醇单胞菌113P3;

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号