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Production strategies for active heme-containing peroxidases from E. coli inclusion bodies – a review

机译:大肠杆菌包涵体生产含活性血红素过氧化物酶的生产策略–综述

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摘要

Heme-containing peroxidases are frequently used in medical applications. However, these enzymes are still extracted from their native source, which leads to inadequate yields and a mixture of isoenzymes differing in glycosylation which limits subsequent enzyme applications. Thus, recombinant production of these enzymes in Escherichia coli is a reasonable alternative. Even though production yields are high, the product is frequently found as protein aggregates called inclusion bodies (IBs). These IBs have to be solubilized and laboriously refolded to obtain active enzyme. Unfortunately, refolding yields are still very low making the recombinant production of these enzymes in E. coli not competitive.Motivated by the high importance of that enzyme class, this review aims at providing a comprehensive summary of state-of-the-art strategies to obtain active peroxidases from IBs. Additionally, various refolding techniques, which have not yet been used for this enzyme class, are discussed to show alternative and potentially more efficient ways to obtain active peroxidases from E. coli.
机译:含血红素的过氧化物酶经常用于医学应用。然而,这些酶仍从其天然来源中提取,这导致产量不足以及糖基化作用不同的同工酶混合物,从而限制了后续酶的应用。因此,在大肠杆菌中重组生产这些酶是合理的选择。即使产量很高,该产品也经常被发现为蛋白质聚集体,称为包涵体(IBs)。这些IB必须被溶解并且费力地重折叠以获得活性酶。不幸的是,重折叠的产量仍然很低,使得这些酶在大肠杆菌中的重组生产没有竞争力。由于该酶类别的高度重要性,本综述旨在提供有关该酶最新技术的全面总结。从IB获得活性过氧化物酶。另外,讨论了尚未用于该酶类别的各种重折叠技术,以显示从大肠杆菌获得活性过氧化物酶的替代的且可能更有效的方法。

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