Production of extracellular heterologous proteins in Streptomyces rimosus, producer of the antibiotic oxytetracycline
Production of extracellular heterologous proteins in Streptomyces rimosus, producer of the antibiotic oxytetracycline
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DOI：
10.1007/s00253-018-8793-z
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发表时间：
2018-03-01
影响因子：
5
通讯作者：
Petkovic, Hrvoje
中科院分区：
文献类型：
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作者：
Carrillo Rincon, Andres Felipe;Magdevska, Vasilka;Petkovic, Hrvoje
在链霉菌中，变铅青链霉菌可以直接将目标蛋白分泌到培养基中，因此常被用来生产外源蛋白。另一方面，链霉菌长期以来一直被用于工业规模的土霉素生产，它保持着“普遍公认的安全”状态。该工业菌株具有许多特性，使其成为异源蛋白生产的理想宿主，包括(1)快速生长；(2)像大肠杆菌一样以短片段的形式生长；(3)电穿孔转化效率高；(4)蛋白质分泌到培养基中。在这项研究中，我们特别致力于探索利用SEC分泌途径在S.rimosus宿主中输出异源蛋白。我们的目标是开发一种核糖链霉菌的遗传工具试剂盒，并评估该工业宿主的目的异源蛋白的胞外生产。本研究表明，里氏链霉菌能够在胞外产生具有工业意义的植酸酶AppA，并且类似于以大肠杆菌为宿主，应用His-Tag/Ni亲和层析为提纯活性植酸酶AppA提供了一种简单、快速的方法。因此，我们证明了金黄色葡萄球菌可以作为一种潜在的替代蛋白表达系统。
Among the Streptomyces species, Streptomyces lividans has often been used for the production of heterologous proteins as it can secrete target proteins directly into the culture medium. Streptomyces rimosus, on the other hand, has for long been used at an industrial scale for oxytetracycline production, and it holds 'Generally Recognised As Safe' status. There are a number of properties of S. rimosus that make this industrial strain an attractive candidate as a host for heterologous protein production, including (1) rapid growth rate; (2) growth as short fragments, as for Escherichia coli; (3) high efficiency of transformation by electroporation; and (4) secretion of proteins into the culture medium. In this study, we specifically focused our efforts on an exploration of the use of the Sec secretory pathway to export heterologous proteins in a S. rimosus host. We aimed to develop a genetic tool kit for S. rimosus and to evaluate the extracellular production of target heterologous proteins of this industrial host. This study demonstrates that S. rimosus can produce the industrially important enzyme phytase AppA extracellularly, and analogous to E. coli as a host, application of His-Tag/Ni-affinity chromatography provides a simple and rapid approach to purify active phytase AppA in S. rimosus. We thus demonstrate that S. rimosus can be used as a potential alternative protein expression system.