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Cloning and Heterologous Expression of a Novel Xylanase Gene TAX1 from Trichoderma atroviride and Its Application in the Deconstruction of Corn Stover

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Abstract

Xylanase plays a vital role in the efficient utilization of xylan, which accounts for up to 30% of plant dry matter. However, the production cost of xylanase remains high, and the enzymatic characteristics of xylanases of most microorganisms are not suitable for industrial production. Therefore, it is of great significance to discover and develop new and efficient xylanases. In this study, the xylanase gene TAX1 (672 bp cDNA) was cloned from Trichoderma atroviride 3.3013 and expressed in Pichia pastoris. The TAX1 gene encoded a 223-amino acid protein (TAX1) with a molecular weight of 24.2 kDa which showed high similarity to glycoside hydrolase family 11. Enzyme activity assay verified that the recombinant xylanase TAX1 had optimal activity (215.3 IU/mL) at 50°C and pH 6.0. Stable working conditions were measured as pH 4.0–7.0 and 40–60°C. By adding Zn2+, the relative enzymatic activity of recombinant TAX1 was enhanced by 26%. The recombinant xylanase showed high activity toward birchwood xylan and corn stover. The Km and Kcat for xylan and corn stover were 0.36 mg/mL and 0.204 S−1 and 0.48 mg/mL and 0.149 S−1, respectively. The enzymatic activity of the TAX1 produced by P. pastoris was about 2.4–4 times higher that directly isolated from T. atroviride, so engineered P. pastoris for xylanase production could be an ideal candidate for industrial enzyme production.

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Funding

This work was partially supported by funds provided by the Natural Science Foundation of Heilongjiang Province (C2015002 and LH2020C011) and the Postdoctoral Scientific Research Developmental Fund of Heilongjiang Province (LBH-Q18023).

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  1. Jin Cai and Xiu-Ling Chen contributed equally to this work.

Authors and Affiliations

  1. Heilongjiang Vocational College of Agricultural Technology, Jiamusi, 154007, People’s Republic of China

    Jin Cai & Xiao-Mei Huang

  2. Northeast Agricultural University, Harbin, 150030, People’s Republic of China

    Xiu-Ling Chen, Jin-Xia Fan, Rui Li, Xu-Dong Sun, Qing-Qing Li & Dong-Yu Li

  3. The College of Engineering, Northeast Agricultural University, Harbin, 150030, People’s Republic of China

    Jin-Xia Fan

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  1. Jin Cai
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Contributions

JC and XC performed research and wrote the paper. JF designed the study and wrote the paper. XH designed the study, provided funding, and edited the paper. RL analyzed the data and edited the paper. XS, QL, and DL performed research.

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Correspondence to Jin-Xia Fan or Xiao-Mei Huang.

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Cai, J., Chen, XL., Fan, JX. et al. Cloning and Heterologous Expression of a Novel Xylanase Gene TAX1 from Trichoderma atroviride and Its Application in the Deconstruction of Corn Stover. Appl Biochem Biotechnol 193, 3029–3044 (2021). https://doi.org/10.1007/s12010-021-03582-0

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