Increasing cell biomass in Saccharomyces cerevisiae increases recombinant protein yield

the use of a respiratory strain as a microbial cell factory

Cecilia Ferndahl, Nicklas Bonander, Christel Logez, Renaud Wagner, Lena Gustafsson, Christer Larsson, Kristina Hedfalk, Richard A.J. Darby, Roslyn M. Bill

Research output: Contribution to journalArticle

Abstract

BACKGROUND: Recombinant protein production is universally employed as a solution to obtain the milligram to gram quantities of a given protein required for applications as diverse as structural genomics and biopharmaceutical manufacture. Yeast is a well-established recombinant host cell for these purposes. In this study we wanted to investigate whether our respiratory Saccharomyces cerevisiae strain, TM6*, could be used to enhance the productivity of recombinant proteins over that obtained from corresponding wild type, respiro-fermentative strains when cultured under the same laboratory conditions. RESULTS: Here we demonstrate at least a doubling in productivity over wild-type strains for three recombinant membrane proteins and one recombinant soluble protein produced in TM6* cells. In all cases, this was attributed to the improved biomass properties of the strain. The yield profile across the growth curve was also more stable than in a wild-type strain, and was not further improved by lowering culture temperatures. This has the added benefit that improved yields can be attained rapidly at the yeast's optimal growth conditions. Importantly, improved productivity could not be reproduced in wild-type strains by culturing them under glucose fed-batch conditions: despite having achieved very similar biomass yields to those achieved by TM6* cultures, the total volumetric yields were not concomitantly increased. Furthermore, the productivity of TM6* was unaffected by growing cultures in the presence of ethanol. These findings support the unique properties of TM6* as a microbial cell factory. CONCLUSIONS: The accumulation of biomass in yeast cell factories is not necessarily correlated with a proportional increase in the functional yield of the recombinant protein being produced. The respiratory S. cerevisiae strain reported here is therefore a useful addition to the matrix of production hosts currently available as its improved biomass properties do lead to increased volumetric yields without the need to resort to complex control or cultivation schemes. This is anticipated to be of particular value in the production of challenging targets such as membrane proteins.
Original languageEnglish
Article number47
Pages (from-to)47
JournalMicrobial Cell Factories
Volume9
DOIs
Publication statusPublished - 17 Jun 2010

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Recombinant proteins
Recombinant Proteins
Yeast
Biomass
Saccharomyces cerevisiae
Industrial plants
Yeasts
Productivity
Membrane Proteins
Proteins
Growth
Genomics
Membranes
Ethanol
Glucose
Temperature
Cells

Bibliographical note

© 2010 Ferndahl et al; licensee BioMed Central Ltd.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Keywords

  • recombinant protein production
  • protein
  • structural genomics
  • biopharmaceutical manufacture
  • yeast
  • respiratory Saccharomyces cerevisiae strain
  • TM6*
  • productivity
  • recombinant proteins
  • respiro-fermentative strains

Cite this

Ferndahl, Cecilia ; Bonander, Nicklas ; Logez, Christel ; Wagner, Renaud ; Gustafsson, Lena ; Larsson, Christer ; Hedfalk, Kristina ; Darby, Richard A.J. ; Bill, Roslyn M. / Increasing cell biomass in Saccharomyces cerevisiae increases recombinant protein yield : the use of a respiratory strain as a microbial cell factory. In: Microbial Cell Factories. 2010 ; Vol. 9. pp. 47.
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Increasing cell biomass in Saccharomyces cerevisiae increases recombinant protein yield : the use of a respiratory strain as a microbial cell factory. / Ferndahl, Cecilia; Bonander, Nicklas; Logez, Christel; Wagner, Renaud; Gustafsson, Lena; Larsson, Christer; Hedfalk, Kristina; Darby, Richard A.J.; Bill, Roslyn M.

In: Microbial Cell Factories, Vol. 9, 47, 17.06.2010, p. 47.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Increasing cell biomass in Saccharomyces cerevisiae increases recombinant protein yield

T2 - the use of a respiratory strain as a microbial cell factory

AU - Ferndahl, Cecilia

AU - Bonander, Nicklas

AU - Logez, Christel

AU - Wagner, Renaud

AU - Gustafsson, Lena

AU - Larsson, Christer

AU - Hedfalk, Kristina

AU - Darby, Richard A.J.

AU - Bill, Roslyn M.

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Y1 - 2010/6/17

N2 - BACKGROUND: Recombinant protein production is universally employed as a solution to obtain the milligram to gram quantities of a given protein required for applications as diverse as structural genomics and biopharmaceutical manufacture. Yeast is a well-established recombinant host cell for these purposes. In this study we wanted to investigate whether our respiratory Saccharomyces cerevisiae strain, TM6*, could be used to enhance the productivity of recombinant proteins over that obtained from corresponding wild type, respiro-fermentative strains when cultured under the same laboratory conditions. RESULTS: Here we demonstrate at least a doubling in productivity over wild-type strains for three recombinant membrane proteins and one recombinant soluble protein produced in TM6* cells. In all cases, this was attributed to the improved biomass properties of the strain. The yield profile across the growth curve was also more stable than in a wild-type strain, and was not further improved by lowering culture temperatures. This has the added benefit that improved yields can be attained rapidly at the yeast's optimal growth conditions. Importantly, improved productivity could not be reproduced in wild-type strains by culturing them under glucose fed-batch conditions: despite having achieved very similar biomass yields to those achieved by TM6* cultures, the total volumetric yields were not concomitantly increased. Furthermore, the productivity of TM6* was unaffected by growing cultures in the presence of ethanol. These findings support the unique properties of TM6* as a microbial cell factory. CONCLUSIONS: The accumulation of biomass in yeast cell factories is not necessarily correlated with a proportional increase in the functional yield of the recombinant protein being produced. The respiratory S. cerevisiae strain reported here is therefore a useful addition to the matrix of production hosts currently available as its improved biomass properties do lead to increased volumetric yields without the need to resort to complex control or cultivation schemes. This is anticipated to be of particular value in the production of challenging targets such as membrane proteins.

AB - BACKGROUND: Recombinant protein production is universally employed as a solution to obtain the milligram to gram quantities of a given protein required for applications as diverse as structural genomics and biopharmaceutical manufacture. Yeast is a well-established recombinant host cell for these purposes. In this study we wanted to investigate whether our respiratory Saccharomyces cerevisiae strain, TM6*, could be used to enhance the productivity of recombinant proteins over that obtained from corresponding wild type, respiro-fermentative strains when cultured under the same laboratory conditions. RESULTS: Here we demonstrate at least a doubling in productivity over wild-type strains for three recombinant membrane proteins and one recombinant soluble protein produced in TM6* cells. In all cases, this was attributed to the improved biomass properties of the strain. The yield profile across the growth curve was also more stable than in a wild-type strain, and was not further improved by lowering culture temperatures. This has the added benefit that improved yields can be attained rapidly at the yeast's optimal growth conditions. Importantly, improved productivity could not be reproduced in wild-type strains by culturing them under glucose fed-batch conditions: despite having achieved very similar biomass yields to those achieved by TM6* cultures, the total volumetric yields were not concomitantly increased. Furthermore, the productivity of TM6* was unaffected by growing cultures in the presence of ethanol. These findings support the unique properties of TM6* as a microbial cell factory. CONCLUSIONS: The accumulation of biomass in yeast cell factories is not necessarily correlated with a proportional increase in the functional yield of the recombinant protein being produced. The respiratory S. cerevisiae strain reported here is therefore a useful addition to the matrix of production hosts currently available as its improved biomass properties do lead to increased volumetric yields without the need to resort to complex control or cultivation schemes. This is anticipated to be of particular value in the production of challenging targets such as membrane proteins.

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