Impact of protein uptake and degradation on recombinant protein secretion in yeast

Keith E. J. Tyo, Zihe Liu, Ylva Magnusson, Dina Petranovic, Jens Nielsen

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Protein titers, a key bioprocessing metric, depend both on the synthesis of protein and the degradation of protein. Secreted recombinant protein production in Saccharomyces cerevisiae is an attractive platform as minimal media can be used for cultivation, thus reducing fermentation costs and simplifying downstream purification, compared to other systems that require complex media. As such, engineering S. cerevisiae to improve titers has been then the subject of significant attention, but the majority of previous efforts have been focused on improving protein synthesis. Here, we characterize the protein uptake and degradation pathways of S. cerevisiae to better understand its impact on protein secretion titers. We do find that S. cerevisiae can consume significant (in the range of 1 g/L/day) quantities of whole proteins. Characterizing the physiological state and combining metabolomics and transcriptomics, we identify metabolic and regulatory markers that are consistent with uptake of whole proteins by endocytosis, followed by intracellular degradation and catabolism of substituent amino acids. Uptake and degradation of recombinant protein products may be common in S. cerevisiae protein secretion systems, and the current data should help formulate strategies to mitigate product loss.
Original languageEnglish
JournalApplied Microbiology and Biotechnology
Volume98
Issue number16
Pages (from-to)7149-7159
ISSN0175-7598
DOIs
Publication statusPublished - 2014

Keywords

  • endocytosis
  • protein degradation pathway
  • protein uptake
  • Fungi Plantae (Fungi, Microorganisms, Nonvascular Plants, Plants) - Ascomycetes [15100] Saccharomyces cerevisiae species yeast common strain-CEN.PK 113-7D
  • amino acid catabolism intracellular degradation
  • metabolic marker
  • recombinant protein
  • regulatory marker
  • 10060, Biochemistry studies - General
  • 10064, Biochemistry studies - Proteins, peptides and amino acids
  • 39008, Food microbiology - General and miscellaneous
  • 51522, Plant physiology - Chemical constituents
  • Biochemistry and Molecular Biophysics
  • Bioprocess Engineering
  • BIOTECHNOLOGY
  • SACCHAROMYCES-CEREVISIAE
  • METABOLOMICS
  • EXTRACTION
  • TOPOLOGY
  • SOFTWARE
  • RATES
  • Endocytosis
  • Saccharomyces cerevisiae
  • Protein degradation
  • Protein production
  • Secretion
  • PROTEIN synthesis
  • HASH(0x34dfcd0)

Cite this

Tyo, Keith E. J. ; Liu, Zihe ; Magnusson, Ylva ; Petranovic, Dina ; Nielsen, Jens. / Impact of protein uptake and degradation on recombinant protein secretion in yeast. In: Applied Microbiology and Biotechnology. 2014 ; Vol. 98, No. 16. pp. 7149-7159.
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Impact of protein uptake and degradation on recombinant protein secretion in yeast. / Tyo, Keith E. J.; Liu, Zihe; Magnusson, Ylva; Petranovic, Dina; Nielsen, Jens.

In: Applied Microbiology and Biotechnology, Vol. 98, No. 16, 2014, p. 7149-7159.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Impact of protein uptake and degradation on recombinant protein secretion in yeast

AU - Tyo, Keith E. J.

AU - Liu, Zihe

AU - Magnusson, Ylva

AU - Petranovic, Dina

AU - Nielsen, Jens

PY - 2014

Y1 - 2014

N2 - Protein titers, a key bioprocessing metric, depend both on the synthesis of protein and the degradation of protein. Secreted recombinant protein production in Saccharomyces cerevisiae is an attractive platform as minimal media can be used for cultivation, thus reducing fermentation costs and simplifying downstream purification, compared to other systems that require complex media. As such, engineering S. cerevisiae to improve titers has been then the subject of significant attention, but the majority of previous efforts have been focused on improving protein synthesis. Here, we characterize the protein uptake and degradation pathways of S. cerevisiae to better understand its impact on protein secretion titers. We do find that S. cerevisiae can consume significant (in the range of 1 g/L/day) quantities of whole proteins. Characterizing the physiological state and combining metabolomics and transcriptomics, we identify metabolic and regulatory markers that are consistent with uptake of whole proteins by endocytosis, followed by intracellular degradation and catabolism of substituent amino acids. Uptake and degradation of recombinant protein products may be common in S. cerevisiae protein secretion systems, and the current data should help formulate strategies to mitigate product loss.

AB - Protein titers, a key bioprocessing metric, depend both on the synthesis of protein and the degradation of protein. Secreted recombinant protein production in Saccharomyces cerevisiae is an attractive platform as minimal media can be used for cultivation, thus reducing fermentation costs and simplifying downstream purification, compared to other systems that require complex media. As such, engineering S. cerevisiae to improve titers has been then the subject of significant attention, but the majority of previous efforts have been focused on improving protein synthesis. Here, we characterize the protein uptake and degradation pathways of S. cerevisiae to better understand its impact on protein secretion titers. We do find that S. cerevisiae can consume significant (in the range of 1 g/L/day) quantities of whole proteins. Characterizing the physiological state and combining metabolomics and transcriptomics, we identify metabolic and regulatory markers that are consistent with uptake of whole proteins by endocytosis, followed by intracellular degradation and catabolism of substituent amino acids. Uptake and degradation of recombinant protein products may be common in S. cerevisiae protein secretion systems, and the current data should help formulate strategies to mitigate product loss.

KW - endocytosis

KW - protein degradation pathway

KW - protein uptake

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KW - amino acid catabolism intracellular degradation

KW - metabolic marker

KW - recombinant protein

KW - regulatory marker

KW - 10060, Biochemistry studies - General

KW - 10064, Biochemistry studies - Proteins, peptides and amino acids

KW - 39008, Food microbiology - General and miscellaneous

KW - 51522, Plant physiology - Chemical constituents

KW - Biochemistry and Molecular Biophysics

KW - Bioprocess Engineering

KW - BIOTECHNOLOGY

KW - SACCHAROMYCES-CEREVISIAE

KW - METABOLOMICS

KW - EXTRACTION

KW - TOPOLOGY

KW - SOFTWARE

KW - RATES

KW - Endocytosis

KW - Saccharomyces cerevisiae

KW - Protein degradation

KW - Protein production

KW - Secretion

KW - PROTEIN synthesis

KW - HASH(0x34dfcd0)

U2 - 10.1007/s00253-014-5783-7

DO - 10.1007/s00253-014-5783-7

M3 - Journal article

VL - 98

SP - 7149

EP - 7159

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

IS - 16

ER -