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首页> 外文期刊>Bioprocess and Biosystems Engineering >Simplified feeding strategies for the fed-batch cultivation of Kluyveromyces lactis GG799 for enhanced recombinant xylanase production
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Simplified feeding strategies for the fed-batch cultivation of Kluyveromyces lactis GG799 for enhanced recombinant xylanase production

机译：乳酸克鲁维酵母GG799分批分批培养的简化饲养策略，可提高重组木聚糖酶的产量

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摘要

A xylanase gene (xyn2) from Trichoderma reesei ATCC 58350 was previously cloned and expressed in Kluyveromyces lactis GG799. The production of the recombinant xylanase was conducted in a developed medium with an optimised batch and with fed-batches that were processed with glucose. The glucose served as a carbon source for cell growth and as an inducer for xylanase production. In a 1-L batch system, a glucose concentration of 20 g L~(-1) and 80 % dissolved oxygen were found to provide the best conditions for the tested ranges. A xylanase activity of 75.53 U mL~(-1) was obtained. However, in the batch mode, glucose depletions reduced the synthesis of recombinant xylanase by K. lactis GG799. To maximise the production of xylanase, further optimisation was performed using exponential feeding. We investigated the effects of various nitrogen sources combined with the carbon to nitrogen (C/N) molar ratio on the production of xylanase. Of the various nitrogen sources, yeast extract was found to be the most useful for recombinant xylanase production. The highest xylanase production (110.13 U mL~(-1)) was measured at a C/N ratio of 50.08. These conditions led to a 45.8 % increase in xylanase activity compared with the batch cultures. Interestingly, the further addition of 500 g L~(-1) glucose led to a 6.2-fold increase (465.07 U mL~(-1)) in recombinant xylanase activity. These findings, together with those of the exponential feeding strategy, indicate that the composition of the C/N molar ratio has a substantial impact on recombinant protein production in K. lactis.
机译：先前克隆了来自里氏木霉ATCC 58350的木聚糖酶基因（xyn2），并在乳酸克鲁维酵母GG799中表达。重组木聚糖酶的生产是在具有优化批次和已用葡萄糖处理的补料分批的发达培养基中进行的。葡萄糖用作细胞生长的碳源和木聚糖酶产生的诱导剂。在1 L分批系统中，发现20 g L〜（-1）的葡萄糖浓度和80％的溶解氧为测试范围提供了最佳条件。获得的木聚糖酶活性为75.53 U mL〜（-1）。然而，在分批模式下，葡萄糖的消耗减少了乳酸克鲁维酵母GG799的重组木聚糖酶的合成。为了最大化木聚糖酶的产量，使用指数进料进行了进一步的优化。我们研究了各种氮源以及碳氮比（C / N）对木聚糖酶生产的影响。在各种氮源中，发现酵母提取物对重组木聚糖酶生产最有用。在50.08的C / N比下测得最高的木聚糖酶产量（110.13 U mL〜（-1））。与分批培养相比，这些条件导致木聚糖酶活性增加了45.8％。有趣的是，进一步添加500 g L〜（-1）葡萄糖导致重组木聚糖酶活性增加6.2倍（465.07 U mL〜（-1））。这些发现以及指数进食策略的发现表明，C / N摩尔比的组成对乳酸克鲁维酵母中重组蛋白的产生具有重大影响。

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  • 来源
    《Bioprocess and Biosystems Engineering》 |2014年第9期|1887-1898|共12页
  • 作者

    Siti Fatimah Zaharah Mohamad Fuzi; Firdausi Razali; Jamaliah Jahim; Roshanida A. Rahman; Rosli Illias;

  • 作者单位

    Technology and Heritage Department, Faculty of Science, Technology and Human Development, Universiti Tun Hussein Onn, 86400 Parit Raja, Batu Pahat, Johor, Malaysia;

    Bioprocess Engineering Department, Faculty of Chemical Engineering, Universiti Teknologi Malaysia, 81310 Skudai, Johor, Malaysia;

    Chemical and Process Engineering Department, Faculty of Engineering and Built Environment, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia;

    Bioprocess Engineering Department, Faculty of Chemical Engineering, Universiti Teknologi Malaysia, 81310 Skudai, Johor, Malaysia;

    Bioprocess Engineering Department, Faculty of Chemical Engineering, Universiti Teknologi Malaysia, 81310 Skudai, Johor, Malaysia;

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  • 正文语种 eng
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    Recombinant xylanase production; K. lactis optimisation; C/N molar ratio; Feeding strategy;

    机译：重组木聚糖酶生产;乳酸克鲁维酵母的优化;C / N摩尔比;喂养策略;

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