Food and Fermentation Industries >

2024 , Vol. 50 >Issue 5: 14 - 21

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.035557

Replacement of cellulose binding domain of Aspergillus niger thermostable glucanase and properties of fusion enzymes

  • SHAN Yilan ,
  • YANG Menglian ,
  • ZHAO Lin ,
  • SHEN Wei ,
  • YANG Haiquan ,
  • XIA Yuanyuan ,
  • CHEN Xianzhong
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  • 1(The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China)
    2(Anhui Huaheng Biotechnology Co.Ltd., Hefei 231131, China)

Received date: 2023-03-21

  Revised date: 2023-05-08

  Online published: 2024-04-09

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Abstract

This work aimed to investigate a novel approach to obtain thermostable glucanases capable of cellulose hydrolysis by replacing the cellulose binding domain (CBD) with carbohydrate binding domain (CBM) from various families.The focus of this study was the AnEglA6 enzyme obtained from Aspergillus niger and has excellent thermostability and potential application in poultry feed production.Fusion genes were constructed by replacing the CBD-encoding region of eglA6 with CBMs and then expressed in Pichia pastoris.The fusion enzyme AnEg-CBM9 was obtained by combining the catalytic domain of AnEglA6 with the CBM of xylanase from the hyperthermophilic bacterium Caldicellulosiruptor kristjansonii.AnEg-CBM10 was composed of eglA6 and CBM of xylanase from Pseudomonas fluorescens.Both fusion enzymes exhibited high cellulose hydrolysis activity and expanded their substrate range by demonstrating certain hydrolysis activity on microcrystalline cellulose.AnEg-CBM10 showed remarkable improvement in thermal stability, with a half-life of approximately 40 min at 85 ℃ and the ability to withstand high temperatures of 90 ℃ for a brief period.Therefore, AnEg-CBM10 is a novel glucanase with improved thermal stability and cellulose hydrolysis activity.

Key words： glucanase; thermal stability; combined domain; CBM9; CBM10

Cite this article

SHAN Yilan , YANG Menglian , ZHAO Lin , SHEN Wei , YANG Haiquan , XIA Yuanyuan , CHEN Xianzhong . Replacement of cellulose binding domain of Aspergillus niger thermostable glucanase and properties of fusion enzymes[J]. Food and Fermentation Industries, 2024 , 50(5) : 14 -21 . DOI: 10.13995/j.cnki.11-1802/ts.035557

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