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Recovery of enzyme byproducts during recombinant protein recovery from transgenic plants

机译:转基因植物重组蛋白质回收过程中酶副产物的回收率

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Recent use of transgenic plants as an alternative protein expression system offers the potential to recover native plant enzymes as byproducts from recombinant protein purification processes. Due to the nature of the native proteins, canola and com are suitable hosts for acidic recombinant proteins, and soybean for the basic ones. Ion-exchange chromatography and polyelectrolyte precipitation as established techniques in recombinant protein downstream processing were applied to non-transgenic plant extracts to assess the feasibility of enzyme byproduct recovery. Cation exchange chromatography of canola and soy extracts completely recovered the native α-galactosidase and urease activities in the extract. α-Gal-actosidase and β-glucosidase activities were fully recovered in anion exchange chromatography of corn germ proteins with enrichment factors of about 3. Simple anion exchange chromatography for canola extracts recovered α-galactosidase activity with 59% yield and 11 times enrichment. Polyelectrolyte precipitation using polyethyleneimine at a dosage of 38 mg/g total protein recovered 47% of α-galactosidase from canola with an enrichment factor of about 3. Several recommendations on the use of the byproduct fractions are proposed. Additional processing of byproduct fractions to concentrate form is economically feasible.
机译:最近使用转基因植物作为替代蛋白表达系统,提供了从重组蛋白质纯化过程中恢复原生植物酶作为副产物的潜力。由于天然蛋白质的性质,CanoLA和COM是酸性重组蛋白的合适宿主,以及碱性的大豆。将离子交换色谱和聚电解质沉淀作为成立的重组蛋白下游处理中的技术应用于非转基因植物提取物,以评估酶副产物回收的可行性。油菜和大豆提取物的阳离子交换色谱完全回收了萃取物的天然α-半乳糖苷酶和脲酶活性。在玉米胚芽蛋白的阴离子交换色谱中完全回收α-加仑曲孔酶和β-葡萄糖苷酶活性,其富集因子约为3.简单的阴离子交换色谱,用于油菜溶液的α-半乳糖苷酶活性,产率为59%和11次富集。使用聚乙烯亚胺的聚乙烯在38mg / g总蛋白质中的聚乙烯沉淀从油菜中回收47%的α-半乳糖苷酶,其富集因子约为3.提出了关于使用副产物级分的若干建议。副产品级数以浓缩形式的额外加工在经济上是可行的。

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