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Bioreactor Controls-III

Strain improvement is a foundational strategy in industrial microbiology aimed at maximizing microbial productivity, particularly because natural isolates typically yield commercially valuable products in very low concentrations. Although optimizing the culture medium and environmental conditions can improve yields, these adjustments are inherently limited by the organism’s genetic potential. As a result, the focus shifts toward genetic modifications to enhance biosynthetic capacity. The...

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Related Experiment Video

Updated: May 20, 2026

Protein Engineering by Yeast Surface Display
05:49

Protein Engineering by Yeast Surface Display

Published on: November 29, 2024

Engineering a robust cell-surface display platform in the multi-stress-tolerant yeast Issatchenkia orientalis.

Yoshiaki Kawahara1, Ryo Nasuno2, Yong-Su Jin3,4

  • 1Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan.

Applied Microbiology and Biotechnology
|May 19, 2026
PubMed
Summary

We engineered the yeast Issatchenkia orientalis to display cellulolytic enzymes on its surface, enabling efficient biomass fermentation. This strain thrives even with lignocellulosic fermentation inhibitors, paving the way for sustainable biofuel production.

Keywords:
Issatchenkia orientalisLignocellulosic biomassMulti-stress toleranceYeast cell-surface displayβ-glucosidase

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Enzymatic Modification and Flow Cytometry Assessment of Yeast Surface Displayed Proteins
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Enzymatic Modification and Flow Cytometry Assessment of Yeast Surface Displayed Proteins

Published on: May 30, 2025

Area of Science:

  • Biotechnology
  • Synthetic Biology
  • Microbial Engineering

Background:

  • Cell surface display of enzymes integrates biomass saccharification and fermentation.
  • Lignocellulosic biomass (LCB) requires robust microbial strains tolerant to harsh pretreatment conditions.
  • Issatchenkia orientalis exhibits natural tolerance to multi-stress conditions.

Purpose of the Study:

  • To establish a cell-surface display system for cellulolytic enzymes in Issatchenkia orientalis.
  • To engineer a strain capable of utilizing LCB-derived sugars under inhibitory conditions.
  • To develop a platform for sustainable LCB bioconversion.

Main Methods:

  • Constructed a modified cell-surface display cassette using the I. orientalis SED1 gene.
  • Displayed fluorescent protein and β-glucosidase (BGL) on the yeast cell surface.
  • Assessed strain growth and cellobiose utilization under stress conditions.

Main Results:

  • Successfully established a functional cell-surface display system in I. orientalis.
  • The engineered strain robustly utilized cellobiose as the sole carbon source.
  • The BGL-displaying strain maintained growth despite the presence of lignocellulosic fermentation inhibitors.

Conclusions:

  • Demonstrated the first successful immobilization of functional proteins on the I. orientalis cell surface.
  • Highlighted the potential of I. orientalis as a robust platform for sustainable LCB bioconversion.
  • Showcased the strain's ability to assimilate LCB-derived intermediates under stress.