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The unique GH5 cellulase member in the extreme halotolerant fungus Aspergillus glaucus CCHA is an endoglucanase with multiple tolerance to salt, alkali and heat: prospects for straw degradation applications

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Abstract

In a halotolerant fungus Aspergillus glaucus CCHA, several functional proteins with stress-tolerant activity have been studied, but no secretory enzymes have been identified yet. The unique GH5 cellulase candidate from A. glaucus, an endoglucanase termed as AgCMCase, was cloned, expressed in the Pichia pastoris system and the purified enzyme was characterized. A large amount of recombinant enzyme secreted by the P. pastoris GS115 strain was purified to homogeneity. The molecular weight of the purified endoglucanase is about 55.0 kDa. The AgCMCase exhibited optimum catalytic activity at pH 5.0 and 55 °C. However, it remained relatively stable at temperatures ranging from 45 to 80 °C and pH ranging from 4.0 to 9.0. In addition, it showed higher activity at extreme NaCl concentrations from 1.0 to 4.0 M, suggesting it is an enzyme highly stable under heat, acid, alkaline and saline conditions. To evaluate the catalytic activity of AgCMCase, the hydrolysis products of rice and corn straws were successfully studied. In conclusion, the AgCMCase is a thermostable and salt-tolerant cellulase with potential for industrial application.

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Abbreviations

NCBI:

The National Center for Biotechnology Information

CAZY:

Carbohydrate-active enzymes

CMCase:

Carboxymethyl cellulase

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

IPTG:

Isopropyl-β-d-thiogalactopyranoside

CMC-Na:

Sodium carboxymethyl cellulose

MES:

2-(N-morpholino)ethanesulfonic acid

HEPES:

4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid

EDTA:

Ethylenediaminetetraacetic acid

K m :

Michaelis–Menten constant

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (NSFC: 31171794, 31370689, 31670141, and 31671972).

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Authors and Affiliations

  1. College of Plant Sciences, Jilin University, Changchun, 130062, People’s Republic of China

    Zhengqun Li, Xue Pei, Ziyu Zhang, Yi Wei, Yanyue Song, Lina Chen, Shouan Liu & Shi-Hong Zhang

  2. The High School Attached to Jilin University, Changchun, 130021, People’s Republic of China

    Ziyu Zhang

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  1. Zhengqun Li
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  2. Xue Pei
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  3. Ziyu Zhang
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  4. Yi Wei
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  7. Shouan Liu
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Correspondence to Shouan Liu or Shi-Hong Zhang.

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Communicated by A. Driessen.

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Li, Z., Pei, X., Zhang, Z. et al. The unique GH5 cellulase member in the extreme halotolerant fungus Aspergillus glaucus CCHA is an endoglucanase with multiple tolerance to salt, alkali and heat: prospects for straw degradation applications. Extremophiles 22, 675–685 (2018). https://doi.org/10.1007/s00792-018-1028-5

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