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High-level expression of Aspergillus niger lipase in Pichia pastoris: Characterization and gastric digestion in vitro

机译:黑曲霉脂肪酶在巴斯德毕赤酵母中的高表达:表征和体外消化

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摘要

The low expression level of acidic lipases from Aspergillus sp. remains a major obstacle for their use in industrial applications. In this study, fusion expression with three fusion partners was investigated to enhance the expression level of an acidic lipase from A. niger (ANL) in Pichia pastoris. When fused with a small ubiquitin-related modifier (SUMO), designated SANL, the highest activity reached 960 +/- 40 U/mL in a 3 L fermenter, which was 1.85-fold higher than that of the parent ANL. SANL exhibited its maximum activity at pH 2.5 and had lower K-m and higher k(cat)/K-m values than those of ANL. In gastrointestinal digestion experiments, SANL was resistant to pepsin and had high hydrolytic activity against triolein from pH 3.0 to 6.0. However, SANL was significantly inhibited by NaTDC above its CMC, which may limit its application for intestinal digestion, but allow it to remain suitable for gastric digestion.
机译:曲霉菌中酸性脂肪酶的低表达水平。仍然是它们在工业应用中使用的主要障碍。在这项研究中,研究了与三种融合伴侣的融合表达,以提高巴斯德毕赤酵母中黑曲霉(ANL)酸性脂肪酶的表达水平。当与称为SANL的小的泛素相关修饰剂(SUMO)融合时,在3 L发酵罐中最高活性达到960 +/- 40 U / mL,比亲本ANL高1.85倍。 SANL在pH 2.5时表现出最大的活性,并且比ANL具有更低的K-m和更高的k(cat)/ K-m值。在胃肠道消化实验中,SANL对胃蛋白酶具有抵抗力,并且在pH 3.0至6.0时对三油精具有很高的水解活性。但是,SANL在其CMC之上受到NaTDC的显着抑制,这可能会限制其在肠道消化中的应用,但仍使其适合胃消化。

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