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Functional Dipeptide Production by Immobilized Enzyme on Yeast Cell Surface
Sejin Geum1, Seoyoung Lee1, Sunghee Kim1
1School of Food Science and Biotechnology, Kyungpook National University, Daegu 41566, Republic of Korea.
Journal of Microbiology and Biotechnology
|March 12, 2026
Summary
This study developed a reusable whole-cell biocatalyst using engineered yeast for efficient L-Alanyl-L-glutamine (Ala-Gln) production. The system offers a cleaner, more stable alternative for industrial peptide synthesis.
Area of Science:
- Biotechnology
- Enzyme Engineering
- Industrial Microbiology
Background:
- L-Alanyl-L-glutamine (Ala-Gln) is a valuable dipeptide with enhanced stability and bioavailability for nutritional applications.
- Whole-cell biocatalysts, particularly those with enzyme cell-surface immobilization, present a sustainable and efficient alternative to chemical synthesis for peptide production.
- Enzyme immobilization on yeast cells enhances enzyme stability and simplifies catalyst recovery, crucial for industrial scalability.
Purpose of the Study:
- To develop a clean and efficient whole-cell biocatalytic system for L-Alanyl-L-glutamine (Ala-Gln) production.
- To engineer a *Saccharomyces cerevisiae* strain for surface display of *Sphingobacterium siyangensis* SY1's α-amino acid ester acyltransferase (SsAET).
- To optimize reaction conditions and evaluate the performance and reusability of the engineered biocatalyst for Ala-Gln synthesis.
Main Methods:
- Engineering *Saccharomyces cerevisiae* for cell-surface display of α-amino acid ester acyltransferase (SsAET).
- Optimization of reaction parameters including pH, time, cell concentration, and substrate ratio (AlaOMe/Gln).
- Evaluation of Ala-Gln production yield, catalyst stability, and reusability through repeated-batch reactions.
Main Results:
- An engineered *S. cerevisiae* strain displaying SsAET was successfully developed for Ala-Gln production.
- Optimal conditions (pH 8.0, 3 h, 20 g DCW/l, AlaOMe/Gln = 1:2) led to a 6.7-fold increase in Ala-Gln production.
- Maximum Ala-Gln concentration of 14.12 mM was achieved, with the biocatalyst retaining over 60% activity after three cycles and stable conversion rates.
Conclusions:
- A clean, efficient, and reusable whole-cell biocatalytic system for Ala-Gln production was established using enzyme cell-surface immobilization.
- The engineered yeast biocatalyst demonstrates significant potential for industrial-scale peptide synthesis.
- This approach offers a sustainable and cost-effective method for producing high-value dipeptides like Ala-Gln.
