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Related Experiment Video

Updated: Apr 3, 2026

Immobilization of Multi-biocatalysts in Alginate Beads for Cofactor Regeneration and Improved Reusability
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Inferring an ancestral alginate lyase for improved stability and high-level expression using fed-batch fermentation.

Hongxiao Mu1, Renquan Guo2, Beichen Zhao3

  • 1School of Life Science and Technology, Wuhan Polytechnic University, Wuhan, China.

Frontiers in Microbiology
|April 2, 2026
PubMed
Summary

Engineered alginate lyase variants (AncAlyA1, AncAlyA2) show enhanced activity and stability. Optimized fermentation in Escherichia coli achieved high enzyme yields, paving the way for industrial applications.

Keywords:
alginate lyaseancestral sequence inferringfed-batch fermentationheterologous expressionpH and thermal stability

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Area of Science:

  • Biotechnology
  • Enzymology
  • Microbial Engineering

Background:

  • Alginate lyases are crucial biocatalysts with industrial potential.
  • Their application is hindered by poor stability and low production yields.
  • Heterologous expression and protein engineering are key strategies for improvement.

Purpose of the Study:

  • To engineer enhanced alginate lyase variants with improved stability and activity.
  • To develop an efficient industrial-scale production strategy for the engineered enzymes.
  • To overcome limitations in current alginate lyase applications.

Main Methods:

  • Heterologous expression of alginate lyase from *Flavobacterium* sp. in *Escherichia coli*.
  • Protein engineering using ancestral sequence inference to create improved variants (AncAlyA1, AncAlyA2).
  • Fed-batch fermentation optimization using glucose and yeast extract for large-scale enzyme production.

Main Results:

  • Engineered variants AncAlyA1 and AncAlyA2 exhibited significantly increased catalytic activities (132.4% and 87.3% higher than wild-type).
  • Both variants demonstrated superior thermal and pH stability, with AncAlyA2 retaining ~60% activity at 60°C and pH 4.0/10.0.
  • Optimized fed-batch fermentation achieved high enzyme activities: 5164.8 U/mL for AncAlyA1 and 4220.1 U/mL for AncAlyA2.

Conclusions:

  • Engineered alginate lyases possess enhanced enzymatic properties suitable for industrial use.
  • An efficient fermentation strategy enables high-level production of these improved biocatalysts.
  • This study presents a viable approach for the industrial-scale manufacturing of alginate lyase.