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

Carrillo Rincon, Andres Felipe (author), Magdevska, Vasilka (author), Kranjc, Luka (author), Fujs, Štefan (author), Müller, Rolf (author), Petković, Hrvoje (author)

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.

• Language: English
• Keywords: Streptomyces rimosus, heterologous protein production, gene expression, gene tools, reporter system, phytase
• Work type: Scientific work
• Typology: 1.01 - Original Scientific Article
• Organization: BF - Biotechnical Faculty
• Publication status: Published
• Publication version: Author Accepted Manuscript
• Number of pages: Str. 2607-2620
• Numbering: Vol. 102, iss. 6
• PID: 20.500.12556/RUL-103067
• UDC: 602.3+602.6+604.4:579.873.7
• ISSN on article: 0175-7598
• DOI: 10.1007/s00253-018-8793-z
• COBISS.SI-ID: 4879480
• Publication date in RUL: 13.09.2018

Record is a part of a journal

• Title: Applied microbiology and biotechnology
• Shortened title: Appl. microbiol. biotechnol.
• Publisher: Springer Nature
• ISSN: 0175-7598
• COBISS.SI-ID: 5136135

Secondary language

• Language: Slovenian
• Keywords: streptomicete, Streptomyces rimosus, sekundarni metaboliti, heterologni proteini, izražanje genov, encimi, fitaze

Projects

• Funder: ARRS - Slovenian Research Agency
• Project number: C3330-17-529038
• Name: Raziskovalci-2.0-UL-BF-529038