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)

Data:	2017
Resum:	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.
Ajuts:	Ministerio de Economía y Competitividad TQ2011-28398-CO2-01 Agència de Gestió d'Ajuts Universitaris i de Recerca 2014/SGR-452
Nota:	Altres ajuts: MAGRAMA/EGO22008
Nota:	Altres ajuts: MAGRAMA/PIE201220E003
Drets:	Aquest material està protegit per drets d'autor i/o drets afins. Podeu utilitzar aquest material en funció del que permet la legislació de drets d'autor i drets afins d'aplicació al vostre cas. Per a d'altres usos heu d'obtenir permís del(s) titular(s) de drets.
Llengua:	Anglès
Document:	Article ; recerca ; Versió acceptada per publicar
Matèria:	Streptomyces lividans ; Heterologous protein production ; Agarase ; SipY mutant strain
Publicat a:	Journal of chemical technology and biotechnology, Vol. 92, isue 1 (Jan. 2017) , ISSN 1097-4660

DOI: 10.1002/jctb.4933

Post-print 16 p, 1.5 MB

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