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Comparison of ADH3 promoter with commonly used promoters for recombinant protein production in Pichia pastoris

机译:ADH3启动子与常用启动子在毕赤酵母中重组蛋白生产的比较

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Recombinant protein production under the control of the P-ADH3 was compared with Pichia pastoris P-AOX1 and PGAR The single-copy-clones expressing Aspergillus niger xylanase (XylB) gene with the three different promoters were tested in shake flask and 5 L fed-batch fermentation processes. Recombinant protein production with PADH3, P-AOX1 and P-GAP were initiated by addition of ethanol, methanol and glucose, respectively in the culture medium. The fermentation process was carried out for 72 h at 30 degrees C, pH 5 and 30% dissolved oxygen. Extracellular protein production yield for PADH3 (3725 U/mL) was higher than for P-AOX1 (2095 U/mL) and PGAP (580 U/mL) at fermentor scale under the conditions tested. These results show that the PADH3 promoter is a promising tool for large scale production of recombinant proteins and can be an alternative to the P-AOX1 and P-GAR. (C) 2016 Elsevier Inc. All rights reserved.
机译:将P-ADH3控制下的重组蛋白生产与巴斯德毕赤酵母P-AOX1和PGAR进行了比较。在摇瓶中测试了表达带有三种不同启动子的黑曲霉木聚糖酶(XylB)基因的单拷贝克隆,并用5 L分批发酵过程。通过分别在培养基中添加乙醇,甲醇和葡萄糖来启动PADH3,P-AOX1和P-GAP的重组蛋白生产。发酵过程在30摄氏度,pH 5和30%溶解氧下进行72小时。在测试条件下,在发酵罐规模下,PADH3(3725 U / mL)的细胞外蛋白产量高于P-AOX1(2095 U / mL)和PGAP(580 U / mL)。这些结果表明,PADH3启动子是大规模生产重组蛋白的有前途的工具,并且可以替代P-AOX1和P-GAR。 (C)2016 Elsevier Inc.保留所有权利。

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