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Production of extracellular heterologous proteins in Streptomyces rimosus, producer of the antibiotic oxytetracycline

Author:
Carrillo Rincón, AndrésFelipe, Magdevska, Vasilka, Kranjc, Luka, Fujs, Štefan, Müller, Rolf, Petković, Hrvoje
Source:
Applied microbiology and biotechnology 2018 v.102 no.6 pp. 2607-2620
ISSN:
0175-7598
Subject:
Escherichia coli, GRAS substances, Streptomyces lividans, Streptomyces rimosus, chromatography, culture media, electroporation, heterologous gene expression, oxytetracycline, phytases, protein secretion, protein synthesis, proteins
Abstract:
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.
Agid:
5900490