Applied Microbiology and Biotechnology

, Volume 103, Issue 3, pp 1289–1298 | Cite as

Structure-based design of agarase AgWH50C from Agarivorans gilvus WH0801 to enhance thermostability

  • Pujuan Zhang
  • Jinru Zhang
  • Lujia Zhang
  • Jianan Sun
  • Yuan Li
  • Lian Wu
  • Jiahai Zhou
  • Changhu Xue
  • Xiangzhao MaoEmail author
Biotechnologically relevant enzymes and proteins


AgWH50C, an exo-β-agarase of GH50 isolated from Agarivorans gilvus WH0801, plays a key role in the enzymatic production of neoagarobiose, which has great application prospect in the cosmetics and pharmaceutical industry. In contrast, the poor thermostability becomes the main obstructive factor of glycoside hydrolase (GH) family 50 agarases, including AgWH50C. Herein, based on the AgWH50C crystal structure, we designed several mutants by a multiple cross-linked rational design protocol used thermostability predicting softwares ETSS, PoPMuSiC, and HotMuSiC. To our surprise, the mutant K621F increased its relative activity by as much as 45% and the optimal temperature increased to 38 °C compared to that of wild-type, AgWH50C (30 °C). The thermostability of K621F also exhibited a substantial improvement. Considering that the gelling temperature of the agarose is higher than 35 °C, K621F can be used to hydrolyze agarose for neoagarobiose production.


Agarase Thermostability Structure Predicting softwares Mutants 


Funding information

This work was supported by the National Natural Science Foundation of China (31471607), Applied Basic Research Program of Qingdao (16-5-1-17-jch), China Postdoctoral Science Foundation (2016M590661), and special Program for Applied Research on Super Computation of the NSFC-Guangdong Joint Fund (the second phase) under Grant No. U1501501.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

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

Supplementary material

253_2018_9540_MOESM1_ESM.pdf (1.8 mb)
ESM 1 (PDF 1840 kb)


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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
  2. 2.State Key Laboratory of Bio-organic and Natural Products Chemistry, Chinese Academy of SciencesShanghai Institute of Organic ChemistryShanghaiChina
  3. 3.Shanghai Engineering Research Center of Molecular Therapeutics and New Drug Department, Department of Chemistry, School of Chemistry and Molecular EngineeringEast China Normal UniversityShanghaiChina
  4. 4.Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and TechnologyQingdaoChina

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