The temporal analysis of yeast exponential phase using shotgun proteomics as a fermentation monitoring technique

Eric L. Huang, Valérie Orsat, Manesh B. Shah, Robert L. Hettich, Nathan C. VerBerkmoes, Mark G. Lefsrud

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

System biology and bioprocess technology can be better understood using shotgun proteomics as a monitoring system during the fermentation. We demonstrated a shotgun proteomic method to monitor the temporal yeast proteome in early, middle and late exponential phases. Our study identified a total of 1389 proteins combining all 2D-LC-MS/MS runs. The temporal Saccharomyces cerevisiae proteome was enriched with proteolysis, radical detoxification, translation, one-carbon metabolism, glycolysis and TCA cycle. Heat shock proteins and proteins associated with oxidative stress response were found throughout the exponential phase. The most abundant proteins observed were translation elongation factors, ribosomal proteins, chaperones and glycolytic enzymes. The high abundance of the H-protein of the glycine decarboxylase complex (Gcv3p) indicated the availability of glycine in the environment. We observed differentially expressed proteins and the induced proteins at mid-exponential phase were involved in ribosome biogenesis, mitochondria DNA binding/replication and transcriptional activator. Induction of tryptophan synthase (Trp5p) indicated the abundance of tryptophan during the fermentation. As fermentation progressed toward late exponential phase, a decrease in cell proliferation was implied from the repression of ribosomal proteins, transcription coactivators, methionine aminopeptidase and translation-associated proteins.

Original languageEnglish
Pages (from-to)5206-5214
Number of pages9
JournalJournal of Proteomics
Volume75
Issue number17
DOIs
StatePublished - Sep 18 2012

Funding

We thank Dr. David Tabb and the Yates Proteomics Laboratory at Scripps Research Institute for DTASelect/Contrast software, the Institute for Systems Biology for proteome bioinformatics tools used in analysis of the MS data, and the ORNL Genome Analysis and System Modeling Group for computational resources for proteomic analysis. The ORNL part of this research was sponsored in part by the U.S. Department of Energy under Contract DE-AC05-00OR22725 with Oak Ridge National Laboratory, managed and operated by UT-Battelle, LLC.

FundersFunder number
U.S. Department of EnergyDE-AC05-00OR22725
Oak Ridge National Laboratory

    Keywords

    • Fermentation monitoring
    • Mass spectrometry
    • MudPIT
    • Process development
    • Saccharomyces cerevisiae
    • Shotgun proteomics

    Fingerprint

    Dive into the research topics of 'The temporal analysis of yeast exponential phase using shotgun proteomics as a fermentation monitoring technique'. Together they form a unique fingerprint.

    Cite this