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Screening, cloning, enzymatic properties of a novel thermostable cellulase enzyme, and its potential application on water hyacinth utilization

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Abstract

Cellulose is the cheapest, natural, renewable organic substance that is used as a carbon source in various fields. Water hyacinth, an aquatic plant rich in cellulose, is often used as a raw material in fuel production. However, natural cellulase can be hardly used in industrial production on account of its low thermal stability and activity. In this study, a metagenomic library was constructed. Then, a new cellulase gene, cel1029, was screened by Congo red staining and expressed in the prokaryotic system. Enzymatic properties of Cel1029 were explored, including optimum temperature and pH, thermal and pH stability, and tolerance against organic solvents, metal ions, and salt solutions. Finally, its ability of degrading water hyacinth was identified and evaluated. Cel1029 displayed high homology with endoglucanase in the glycoside hydrolase family 5 (GH5) and had high stability across a broad temperature range. More than 86% of its enzymatic activities were retained between 4 and 60 °C after 24 h of incubation. Single-factor analysis and orthogonal design were further conducted to determine the optimal conditions for the highest reducing sugar yield of water hyacinth. Interestingly, Cel1029 efficiently transformed water hyacinth with a reducing sugar yield of 430.39 mg/g in 22 h. These findings may open the door for significant industrial applications of a novel GH5 cellulase (NCBI Reference Sequence: MK051001, Cel1029) and help identify more efficient methods to degrade cellulose-rich plants.

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Acknowledgements

This work was financially supported by the Guangzhou Basic Clean Cosmetics Manufacturing Co. Ltd.

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This research has been funded by Natural Science Foundation of China (31400680), Science and Technology Plan Project of Guangzhou (201802030009), the Innovation and Strengthening School Project from Guangdong Pharmaceutical University (2016KTSCX067 and 2016SFKC_28), Science and Technology Plan Project of Guangdong Province (2017A010105011, 2014A020208134, 2014A020212602), and Education Project of Guangdong Province (2013KJCX0107).

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Authors and Affiliations

  1. School of Life Sciences and Biopharmaceuticals, Guangdong Pharmaceutical University, Guangzhou, 511400, China

    Xiaoshen Zhao, Zujun Deng & He Li

  2. Guangdong Key Laboratory of Bioactive Drug Research, Guangzhou, 511400, China

    Xiaoshen Zhao, Zujun Deng & He Li

  3. School of Business Medicine, Guangdong Pharmaceutical University, Guangzhou, 511400, China

    Liyang Liu

  4. Guangzhou Basic Clean Cosmetics Manufacturing Co., Ltd, Guangzhou, 510080, China

    Shan Liu, Jeonyun Yun & Xiong Xiao

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  1. Xiaoshen Zhao
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Conceptualization, X.S.Z.; methodology, J.Y.; software, Z.J.D.; validation, X.S.Z., S.L., and J.Y.; formal analysis, X.S.Z.; investigation, Z.J.D.; resources, Z.J.D.; data curation, X.X.; writing—original draft preparation, X.S.Z.; writing—review and editing, L.Y.L.; visualization, X.X.; supervision, H.L.; project administration, H.L.; funding acquisition, S.L.. All authors have read and agreed to the published version of the manuscript.

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Zhao, X., Liu, L., Deng, Z. et al. Screening, cloning, enzymatic properties of a novel thermostable cellulase enzyme, and its potential application on water hyacinth utilization. Int Microbiol 24, 337–349 (2021). https://doi.org/10.1007/s10123-021-00170-4

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