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Engineering polar protein-based biomolecular condensates for modular biosynthesis.

Buhan Yao1, Jingyu Zhou2, Xinyue Su2

  • 1School of Life Sciences, Anhui University, Hefei, 230601, China; Key Laboratory of Human Microenvironment and Precision Medicine of Anhui Higher Education Institutes, Anhui University, Hefei, 230601, Anhui, China; Anhui Province Joint Construction Discipline Key Laboratory of Nanobody Technology, Hefei, China; Anhui Healcurer Heath Biotech Co., Ltd. - Anhui University Joint Postgraduate Training Base of Anhui Province, Hefei, China; Institute of Biochemistry and Microbiology, Anhui University, Hefei, China.

International Journal of Biological Macromolecules
|December 30, 2025
PubMed
Summary

Engineering biomolecular condensates spatially isolates enzymatic reactions, significantly boosting the production of valuable compounds like Lacto-N-tetraose (LNT). This method enhances metabolic flux and yield compared to traditional free enzyme systems.

Keywords:
Biomolecular condensatesFermentationLacto-N-tetraosePolar protein

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Area of Science:

  • Synthetic biology
  • Metabolic engineering
  • Biocatalysis

Background:

  • Microbiology offers sustainable production of chemicals and pharmaceuticals.
  • Uncontrolled enzymes lead to intermediate accumulation and low product yields.

Purpose of the Study:

  • To engineer biomolecular condensates for spatial isolation of enzymatic reactions.
  • To establish an efficient cascade catalytic system using engineered condensates.
  • To enhance modular metabolic assembly and optimize catalytic performance.

Main Methods:

  • Engineered PodJIDR polar protein-based biomolecular condensates.
  • Recruitment of enzymes via peptide pairing or direct fusion.
  • Systematic pathway module refinement by adjusting plasmid copy numbers.
  • Utilized Lacto-N-tetraose (LNT) biosynthesis pathway as a model system.

Main Results:

  • Achieved spatial isolation of enzymatic reactions within synthetic compartments.
  • Significantly improved LNT production efficiency, with yields increasing 4.71-fold over wild-type and 1.79-fold over optimized free enzymes.
  • Demonstrated a corresponding increase in metabolic flux.
  • Validated the potential of biomolecular condensates in multienzyme catalysis.

Conclusions:

  • Biomolecular condensates enhance targeted metabolic pathway efficiency.
  • This technique provides a strategy for reinforcing metabolic flux regulation.
  • Engineered condensates improve product titers and offer a novel approach for biocatalysis.