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Title: Improvement of extracellular bacterial protein production in Pichia pastoris by co-expression of endoplasmic reticulum residing GroEL–GroES
Authors: Pijug Summpunn
Juntratip Jomrit
Watanalai Panbangred
Walailak University
Mahidol University
Keywords: Biochemistry, Genetics and Molecular Biology;Chemical Engineering;Immunology and Microbiology
Issue Date: 1-Mar-2018
Citation: Journal of Bioscience and Bioengineering. Vol.125, No.3 (2018), 268-274
Abstract: © 2017 The Society for Biotechnology, Japan Pichia pastoris is an established host system for heterologous protein expression. However, the potential productivity of this system can be limited. In this study, the Escherichia coli chaperones (GroES–GroEL) were expressed from the PGAP promoter and targeted to the secretory pathway through the endoplasmic reticulum (ER). The ability of the ER targeted chaperones to improve production of bacterial protein in P. pastoris was evaluated. The chaperones tagged with α-factor secretion- and ER retention-signal sequences were co-expressed with either extracellularly secreted phytase or intracellular D-phenylglycine aminotransferase (D-PhgAT) enzymes. The ER residing GroEL–GroES successfully increased the levels of active phytase extracellularly, 1.5–2.3-fold higher than the phytase expression alone, but did not enhance the formation of active, intracellular D-PhgAT. These results indicate that the chaperones have the potential to enhance production of active enzymes when present in the same trafficking pathway. This is the first report on the improvement of extracellular bacterial protein production through co-expression with ER residing bacterial chaperones in the Pichia system. The modified P. pastoris expression system may be beneficial for extracellular expression of other prokaryotic proteins.
ISSN: 13474421
Appears in Collections:Scopus 2018

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