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Background <p><i>Escherichia coli</i> is a well-known host species for studies on recombinant protein expression, typically induced during the exponential phase. Nutrient depletion and the accumulation of toxic substances, which are common during the stationary phase, negatively impact recombinant protein expression, thereby limiting its suitability for efficient protein production. This study aimed to explore a physiological state distinct from conventional strategies for a complementary approach by investigating recombinant protein expression during the stationary phase.</p> Results <p>An unconventional stationary-like state was engineered using glucose to trigger acetate-mediated growth arrest and reduce tryptone to prevent regrowth. Under these conditions, the absolute and relative expression of signal peptide-fused human interferon gamma (mCsn2-SP-hIFN-γ) increased &gt; 2.4 and 2.5-fold, respectively, compared with that in exponential-phase induction.</p> Conclusions <p>The present findings suggest that regardless of the unfavorable conditions of the stationary phase for recombinant protein production, glucose utilization may enable the establishment of a stationary-phase model to address these limitations. Overall, these findings underscore the importance of the intracellular metabolic state and expression timing in optimizing recombinant protein production. Although only a single protein was examined in this study, the results highlight the potential of the unconventional stationary phase as an effective new strategy for enhancing recombinant protein expression.</p>

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A novel stationary phase approach for improved recombinant protein expression in Escherichia coli: beyond the exponential phase

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年2月19日

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  • Yoon-Hyeok Kang,
  • Yong-Kyun Ryu,
  • Jaewon Lee,
  • Eunyoung Jo,
  • Yehui Gang,
  • Taeho Kim,
  • Gun-Hoo Park,
  • Chulhong Oh

摘要

Background

Escherichia coli is a well-known host species for studies on recombinant protein expression, typically induced during the exponential phase. Nutrient depletion and the accumulation of toxic substances, which are common during the stationary phase, negatively impact recombinant protein expression, thereby limiting its suitability for efficient protein production. This study aimed to explore a physiological state distinct from conventional strategies for a complementary approach by investigating recombinant protein expression during the stationary phase.

Results

An unconventional stationary-like state was engineered using glucose to trigger acetate-mediated growth arrest and reduce tryptone to prevent regrowth. Under these conditions, the absolute and relative expression of signal peptide-fused human interferon gamma (mCsn2-SP-hIFN-γ) increased > 2.4 and 2.5-fold, respectively, compared with that in exponential-phase induction.

Conclusions

The present findings suggest that regardless of the unfavorable conditions of the stationary phase for recombinant protein production, glucose utilization may enable the establishment of a stationary-phase model to address these limitations. Overall, these findings underscore the importance of the intracellular metabolic state and expression timing in optimizing recombinant protein production. Although only a single protein was examined in this study, the results highlight the potential of the unconventional stationary phase as an effective new strategy for enhancing recombinant protein expression.