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A Streptomyces lividans SipY defficient strain as a host for protein production : standardization of operational alternatives for model proteins
Vidal Gabarró, Marcel·la (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Gullón, Sonia (Centro Nacional de Biotecnología)
López Vicente, Rebeca (Centro Nacional de Biotecnología)
Caminal i Saperas, Glòria (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Pérez Mellado, Rafael (Centro Nacional de Biotecnología)
López Santín, Josep (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)

Date: 2017
Abstract: 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.
Note: Número d'acord de subvenció MINECO/TQ2011-28398-CO2-01
Note: Número d'acord de subvenció MAGRAMA/EGO22008
Note: Número d'acord de subvenció MAGRAMA/PIE201220E003
Note: Número d'acord de subvenció AGAUR/2014/SGR-452
Rights: Tots els drets reservats.
Language: Anglès.
Document: article ; recerca ; acceptedVersion
Subject: Streptomyces lividans ; Heterologous protein production ; Agarase ; SipY mutant strain
Published in: Journal of chemical technology and biotechnology, Vol. 92, isue 1 (Jan. 2017) , ISSN 0268-2575

DOI: 10.1002/jctb.4933

16 p, 1.5 MB

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Articles > Research articles
Articles > Published articles

 Record created 2016-10-28, last modified 2019-02-05

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