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SHR Neuro Cancer Cardio Lipid Metab Microb
Krainer, FW; Gerstmann, MA; Darnhofer, B; Birner-Gruenberger, R; Glieder, A.
Biotechnological advances towards an enhanced peroxidase production in Pichia pastoris.
J Biotechnol. 2016; 233(31):181-189
Doi: 10.1016/j.jbiotec.2016.07.012
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- Co-authors Med Uni Graz
- Birner-Grünberger Ruth
- Darnhofer Barbara
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- Abstract:
- Horseradish peroxidase (HRP) is a high-demand enzyme for applications in diagnostics, bioremediation, biocatalysis and medicine. Current HRP preparations are isolated from horseradish roots as mixtures of biochemically diverse isoenzymes. Thus, there is a strong need for a recombinant production process enabling a steady supply with enzyme preparations of consistent high quality. However, most current recombinant production systems are limited at titers in the low mg/L range. In this study, we used the well-known yeast Pichia pastoris as host for recombinant HRP production. To enhance recombinant enzyme titers we systematically evaluated engineering approaches on the secretion process, coproduction of helper proteins, and compared expression from the strong methanol-inducible PAOX1 promoter, the strong constitutive PGAP promoter, and a novel bidirectional promoter PHTX1. Ultimately, coproduction of HRP and active Hac1 under PHTX1 control yielded a recombinant HRP titer of 132mg/L after 56h of cultivation in a methanol-independent and easy-to-do bioreactor cultivation process. With regard to the many versatile applications for HRP, the establishment of a microbial host system suitable for efficient recombinant HRP production was highly overdue. The novel HRP production platform in P. pastoris presented in this study sets a new benchmark for this medically relevant enzyme. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.
- Find related publications in this database (using NLM MeSH Indexing)
- Bioreactors -
- Biotechnology - methods
- Horseradish Peroxidase - chemistry
- Horseradish Peroxidase - genetics
- Horseradish Peroxidase - isolation & purification
- Horseradish Peroxidase - metabolism
- Methanol -
- Pichia - genetics
- Pichia - metabolism
- Recombinant Proteins - chemistry
- Recombinant Proteins - genetics
- Recombinant Proteins - isolation & purification
- Recombinant Proteins - metabolism
- Find related publications in this database (Keywords)
- Pichia pastoris
- Recombinant protein production
- Plant peroxidase
- Enzyme secretion
- Unfolded protein response
- Bidirectional promoter