Genome-scale analysis of the high-efficient protein secretion system of Aspergillus oryzae

Lifang Liu, Amir Feizi, Tobias Osterlund, Carsten Hjort, Jens Nielsen

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Abstract

Background: The koji mold, Aspergillus oryzae is widely used for the production of industrial enzymes due to its particularly high protein secretion capacity and ability to perform post-translational modifications. However, systemic analysis of its secretion system is lacking, generally due to the poorly annotated proteome. Results: Here we defined a functional protein secretory component list of A. oryzae using a previously reported secretory model of S. cerevisiae as scaffold. Additional secretory components were obtained by blast search with the functional components reported in other closely related fungal species such as Aspergillus nidulans and Aspergillus niger. To evaluate the defined component list, we performed transcriptome analysis on three a-amylase over-producing strains with varying levels of secretion capacities. Specifically, secretory components involved in the ER-associated processes (including components involved in the regulation of transport between ER and Golgi) were significantly up-regulated, with many of them never been identified for A. oryzae before. Furthermore, we defined a complete list of the putative A. oryzae secretome and monitored how it was affected by overproducing amylase. Conclusion: In combination with the transcriptome data, the most complete secretory component list and the putative secretome, we improved the systemic understanding of the secretory machinery of A. oryzae in response to high levels of protein secretion. The roles of many newly predicted secretory components were experimentally validated and the enriched component list provides a better platform for driving more mechanistic studies of the protein secretory pathway in this industrially important fungus.
Original languageEnglish
JournalB M C Systems Biology
Volume8
Issue number73
ISSN1752-0509
DOIs
Publication statusPublished - 2014

Keywords

  • Aspergillus oryzae
  • Batch Cell Culture Techniques
  • Fungal Proteins
  • Genome, Fungal
  • Genomics
  • Transcription, Genetic
  • alpha-Amylases
  • EC 3.2.1.1 alpha-Amylases
  • post translational modification
  • Fungi Plantae (Fungi, Microorganisms, Nonvascular Plants, Plants) - Ascomycetes [15100] Saccharomyces cerevisiae species
  • Fungi Plantae (Fungi, Microorganisms, Nonvascular Plants, Plants) - Fungi Imperfecti or Deuteromycetes [15500] Aspergillus niger species Aspergillus nidulans species Aspergillus oryzae species
  • alpha-amylase 9000-90-2 EC 3.2.1.1
  • functional protein secretory component
  • gene
  • 03502, Genetics - General
  • 03504, Genetics - Plant
  • 10802, Enzymes - General and comparative studies: coenzymes
  • 20504, Nervous system - Physiology and biochemistry
  • Biochemistry and Molecular Biophysics
  • endoplasmic reticulum
  • Golgi-Mazzoni corpuscle nervous system
  • genome-scale analysis laboratory techniques, genetic techniques
  • transcriptome analysis laboratory techniques, genetic techniques
  • Molecular Genetics
  • MATHEMATICAL
  • ENDOPLASMIC-RETICULUM
  • TRANSCRIPTIONAL ACTIVATOR
  • SACCHAROMYCES-CEREVISIAE
  • FILAMENTOUS FUNGI
  • GENE-EXPRESSION
  • STRESS-RESPONSE
  • AMYLASE GENES
  • CELL-WALLS
  • NIGER
  • INDUCTION
  • alpha-amylase
  • Comparative genomics
  • Secretory pathway
  • Transcriptome

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