A Streptomyces lividans SipY deficient strain as a host for protein production: standardization of operational alternatives for model proteins

Marcel·la V. Gabarró, Sonia Gullón, Rebeca L. Vicente, Glòria Caminal, Rafael P. Mellado, Josep López-Santín

Research output: Contribution to journalArticleResearchpeer-review

6 Citations (Scopus)

Abstract

© 2016 Society of Chemical Industry BACKGROUND: Extracellular protein production by Gram-positive bacteria, such as Streptomyces, may be complementary to current established protein production processes. The performance of a Streptomyces lividans mutant strain, deficient in the major signal peptidase (SipY) is investigated for the production of proteins secreted via the secondary Tat pathway. RESULTS: The SipY deficient strain has shown advantages over the wild type strain, in terms of extracellular productivity, specific activity and rheological behaviour. Two operational modes, batch and fed-batch, have been studied using mannitol as carbon source. The results showed that two successive mannitol additions in fed-batch mode led to improved secretory protein production using Streptomyces agarase as a model protein. This production process was also explored for the Tat secretory protein S. lividans laccase. The predicted sequence for the pre-laccase coding sequence has been cloned into the mutant strain under the control of the agarase promoter. Batch and fed-batch laccase production, using either mannitol or glucose as carbon source, have been developed and quantified. CONCLUSIONS: The usefulness of a Streptomyces lividans SipY deficient strain as protein producer has been demonstrated. A proposed operating mode with substrate additions has been employed for the optimisation of Tat proteins production, although some adjustments might be necessary depending on the secretory protein. © 2016 Society of Chemical Industry.
Original languageEnglish
Pages (from-to)217-223
JournalJournal of Chemical Technology and Biotechnology
Volume92
DOIs
Publication statusPublished - 1 Jan 2017

Keywords

  • SipY mutant strain
  • Streptomyces lividans
  • agarase
  • heterologous protein production

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