Metabolic flux profiling of recombinant protein secreting Pichia pastoris growing on glucose:methanol mixtures.

Microbial Cell Factories
Joel JordàP Ferrer

Abstract

The methylotrophic yeast Pichia pastoris has emerged as one of the most promising yeast hosts for the production of heterologous proteins. Mixed feeds of methanol and a multicarbon source instead of methanol as sole carbon source have been shown to improve product productivities and alleviate metabolic burden derived from protein production. Nevertheless, systematic quantitative studies on the relationships between the central metabolism and recombinant protein production in P. pastoris are still rather limited, particularly when growing this yeast on mixed carbon sources, thus hampering future metabolic network engineering strategies for improved protein production. The metabolic flux distribution in the central metabolism of P. pastoris growing on a mixed feed of glucose and methanol was analyzed by Metabolic Flux Analysis (MFA) using 13C-NMR-derived constraints. For this purpose, we defined new flux ratios for methanol assimilation pathways in P. pastoris cells growing on glucose:methanol mixtures. By using this experimental approach, the metabolic burden caused by the overexpression and secretion of a Rhizopus oryzae lipase (Rol) in P. pastoris was further analyzed. This protein has been previously shown to trigger the unfo...Continue Reading

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Citations

Mar 8, 2014·Microbial Cell Factories·Diethard MattanovichBrigitte Gasser
Feb 5, 2013·Future Microbiology·Brigitte GasserDiethard Mattanovich
May 18, 2013·Biotechnology Advances·Yoshihiro Toya, Hiroshi Shimizu
Sep 1, 2015·Current Opinion in Biotechnology·Allison G McAteeJamey D Young
Nov 29, 2013·Biotechnology and Bioengineering·Hyun Tae HwangArvind Varma
Mar 17, 2016·Analytical Chemistry·Douglas McCloskeyAdam M Feist
Mar 30, 2016·Applied Microbiology and Biotechnology·Justyna NoconDiethard Mattanovich
Sep 25, 2015·BMC Biology·Hannes RußmayerBrigitte Gasser
Dec 30, 2014·Biotechnology and Bioengineering·José Manuel BarrigonJosé Luis Montesinos
May 24, 2014·Metabolic Engineering·Justyna NoconDiethard Mattanovich
Jan 7, 2015·Journal of Industrial Microbiology & Biotechnology·Tobias KleinElmar Heinzle
Sep 5, 2014·FEMS Yeast Research·Jan FörsterLars M Blank
Dec 26, 2015·Biotechnology and Bioengineering·Elena CámaraPau Ferrer
Jun 30, 2016·Biotechnology for Biofuels·Tiina M PakulaMikko Arvas
Mar 3, 2017·ACS Synthetic Biology·Ulrike ObstVolker Sieber
Jan 24, 2015·Nature Reviews. Drug Discovery·Alan L HarveyRonald J Quinn
Dec 8, 2019·World Journal of Microbiology & Biotechnology·Jiachen ZhaoYingping Zhuang
Sep 22, 2017·FEMS Yeast Research·Richard J ZahrlBrigitte Gasser
Jul 17, 2020·Frontiers in Bioengineering and Biotechnology·Miguel Angel Nieto-TaypeFrancisco Valero

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Methods Mentioned

BETA
protein folding
PCR
NMR
nuclear magnetic resonance

Software Mentioned

Varian
VNMR
FCAL
METAFoR
CROMELEON

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