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Carbon and nitrogen sources modulate lipase production in the yeast Yarrowia lipolytica

机译:碳和氮源调节酵母解脂耶氏酵母中脂肪酶的产生

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摘要

Aims: To analyse the influence of nitrogen and carbon sources on extracellular lipase production by Yarrowia lipolytica-overproducing mutant in order to optimize its production in large-scale bioreactors. Methods and Results: The level of lipase production and LIP2 induction, measured using an LIP2-LacZ reporter gene, were compared for different carbon and nitrogen sources and for different concentrations. The localization of the enzyme during growth was also determined by Western blotting analysis using a six-histidine-tagged lipase. Significance and Impact of the Study: Tryptone N1 and oleic acid are the most suitable nitrogen and carbon sources for the production of the extracellular lipase by the Y. lipolytica mutant. Higher levels of lipase production were obtained as the tryptone concentration increased in the culture medium. Such a positive correlation was not observed with oleic acid media where the highest lipolytic productivities were obtained in the presence of low concentration. We also demonstrate that in the presence of oleic acid, lipase is cell-bound during the growth phase before being released in the media. Conclusions: This work provides a better understanding of the mechanism controlling LIP2 expression and, thus, extracellular lipase production in the yeast Y. lipolytica.
机译:目的:分析氮源和碳源对解脂耶氏酵母过量生产突变体生产胞外脂肪酶的影响,以优化其在大型生物反应器中的生产。方法和结果:比较了使用LIP2-LacZ报告基因测量的脂肪酶产生水平和LIP2诱导水平,比较了不同的碳源和氮源以及不同的浓度。还使用六组氨酸标记的脂肪酶通过蛋白质印迹分析确定了生长过程中酶的定位。研究的意义和影响:胰蛋白N N1和油酸是解脂耶氏酵母突变体生产细胞外脂肪酶的最合适氮和碳源。随着培养基中胰蛋白concentration浓度的增加,获得更高水平的脂肪酶生产。在油酸介质中未观察到这样的正相关性,在低浓度的油酸介质中,脂解效率最高。我们还证明,在油酸存在下,脂肪酶在生长阶段会被细胞结合,然后释放到培养基中。结论:这项工作提供了对控制LIP2表达的机制的更好理解,从而更好地理解了解脂耶氏酵母中细胞外脂肪酶的产生。

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