Food and Fermentation Industries >

2023 , Vol. 49 >Issue 4: 216 - 223

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.031705

Molecular truncation of a hyperthermophilic glucanase from Aspergillus niger and optimization of its preparation conditions

  • WU Shangwu ,
  • YANG Menglian ,
  • SHEN Wei ,
  • SHAN Yilan ,
  • WANG Shilan ,
  • YANG Haiquan ,
  • XIA Yuanyuan ,
  • CHEN Xianzhong
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  • 1(School of Biotechnology, Key Laboratory of Industrial Biotechnology, Jiangnan University, Wuxi 214122, China)
    2(Wuxi AppTec New Drug Development Co. Ltd., Suzhou 215000, China)

Received date: 2022-03-25

  Revised date: 2022-04-25

  Online published: 2023-03-20

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Abstract

This paper aims to increase the expression level of hyperthermophilic β-glucanase AnEglA6 from Aspergillus niger molecular truncation, and establish the fermentation conditions for the recombinant strain. Hyperthermophilic β-1,4-glucanase plays an important role in poultry feed production. The glucanase AnEglA6 from Aspergillus niger has superior heat resistance, and it is suitable for use in the poultry feed industry. The molecular weight of glucanase AnEglA6 was reduced by C-terminal truncation, and the gene segment, which encoded the cellulose-binding domain and linker peptide, was removed from the gene eglA6. The truncated gene eglA6c was expressed in Pichia pastoris and Kluyveromyces lactis, and the expression products were designated as AnEglA6cP and AnEglA6cK, respectively. The recombinant enzyme AnEglA6cK, which was expressed by K. lactis, showed highest activity at 75 ℃, and its half-life reached 55 min at 80 ℃. In addition, the enzyme can withstand high temperature of 85 ℃ for a short time. The application performance of AnEglA6cK was evidently better than that of AnEglA6cP. The enzymatic activity of the truncated enzyme AnEglA6cK decreased rapidly in the later stage of fermentation; thus, the organic nitrogen source of the culture medium was optimized. The growth time of the enzymatic activity was prolonged by replacing casein peptone with soybean peptone, and the highest enzymatic activity of fermentation broth in a shake flask was greatly improved.

Key words： hyperthermophilic β-1,4-glucanase; AnEglA6; molecular truncation; Kluyveromyces lactis; soybean peptone

Cite this article

WU Shangwu , YANG Menglian , SHEN Wei , SHAN Yilan , WANG Shilan , YANG Haiquan , XIA Yuanyuan , CHEN Xianzhong . Molecular truncation of a hyperthermophilic glucanase from Aspergillus niger and optimization of its preparation conditions[J]. Food and Fermentation Industries, 2023 , 49(4) : 216 -223 . DOI: 10.13995/j.cnki.11-1802/ts.031705

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