Applied Microbiology and Biotechnology

, Volume 103, Issue 2, pp 807–817 | Cite as

Study on expression and action mode of recombinant alginate lyases based on conserved domains reconstruction

  • Min Yang
  • Nannan Li
  • Suxiao Yang
  • Yuan Yu
  • Zhenlian Han
  • Li LiEmail author
  • Haijin MouEmail author
Biotechnologically relevant enzymes and proteins


Understanding the effect of conserved domains reconstruction of alginate lyases on action mode is essential for their application and in-depth study. We report the expression and action mode of recombinant alginate lyase (AlyM) and its conserved domain reconstruction forms (AlyMΔCBM, cAlyM, and AlyMΔ58C). The enzymatic activities of AlyM, AlyMΔCBM, cAlyM, and AlyMΔ58C were 61.77, 150.57, 388.97, and 308.21 U/mg, respectively. The transcription level of cAlyM was 49.89-fold of AlyM. cAlyM and AlyMΔ58C showed higher thermal stability than AlyM, indicating that the removal of F5_F8_type_C domain was beneficial for the increase of thermal stability. The enzymes were bifunctional alginate lyases and preferred polyG to polyM. The enzymes degraded alginate to produce unsaturated disaccharide, trisaccharide, and tetrasaccharide as the main end-products. Pentamannuronic acid and pentaguluronic acid were the smallest substrates that could be degraded by AlyM, with unsaturated trisaccharide/tetrasaccharide (40.61%/44.42%) and disaccharide/trisaccharide (10.57%/83.85%) as the main products, respectively. The action modes of enzymes remain unaffected after conserved domain reconstruction, but the affinity of AlyMΔ58C toward polyM increased. This study provides a new strategy for rational modification of alginate lyase based on conserved domain reconstruction.


Alginate lyase Action mode Conserved domain reconstruction Transcription level Thermal stability Microbulbifer 


Funding information

This work was supported by the Shandong Province Key Research and Development Project (2017YYSP003), Natural Science Foundation of Shandong Province (ZR2017MD006), and Public Science and Technology Research Funds Projects of Ocean (201505022).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement

This article does not contain any studies with human participants or animals performed by any of the authors.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.College of Food Science and EngineeringOcean University of ChinaQingdaoChina

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