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Selection of new over-producing derivatives for the improvement of extracellular lipase production by the non-conventional yeast Yarrowia lipolytica

机译:非传统酵母解脂耶氏酵母的新过量生产衍生物的选择,以改善细胞外脂肪酶的生产

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The non-conventional yeast Yarrowia lipolytica produces an extracellular lipase encoded by the LIP2 gene. Mutant strains with enhanced productivity were previously obtained either by chemical mutagenesis or genetic engineering. In this work, we used one of these mutants, named LgX64.81 to select new overproducing strains following by amplification of the LIP2 gene. We also developed a process for lipase production in bioreactors and compared lipase production levels in batch and fed-batch cultures. Batch culture led to a lipase production of 26450 U ml(-1) in a media containing olive oil and tryptone as carbon and nitrogen sources. Feeding of a combination of tryptone and olive oil at the end of the exponential growth phase yielded to lipase activity of 158246 U ml(-1) after 80 h of cultivation. In addition this production system developed for the extracellular lipase could also be applied for other heterologous protein production since we have demonstrated that LgX64.81 is an interesting alternative host strain.
机译:非常规酵母解脂耶氏酵母产生由LIP2基因编码的细胞外脂肪酶。先前通过化学诱变或基因工程获得了具有提高的生产率的突变菌株。在这项工作中,我们使用了一个名为LgX64.81的突变体,通过扩增LIP2基因来选择新的高产菌株。我们还开发了一种在生物反应器中生产脂肪酶的方法,并比较了分批和补料分批培养物中脂肪酶的生产水平。分批培养导致在包含橄榄油和胰蛋白as作为碳和氮源的培养基中产生脂肪酶26450 U ml(-1)。培养80小时后,在指数生长期结束时喂入胰蛋白and和橄榄油的混合物,脂肪酶活性为158246 U ml(-1)。另外,为细胞外脂肪酶开发的这种生产系统也可以用于其他异源蛋白的生产,因为我们已经证明LgX64.81是一种有趣的替代宿主菌株。

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