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Purification and characterization of the low molecular weight xylanase from Bacillus cereus L-1

  • Environmental Microbiology - Research Paper
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Abstract

Xylanase is widely used in various industries such as food processing, paper, textiles, and leather tanning. In this study, Bacillus cereus L-1 strain was isolated and identified as capable of producing low molecular weight xylanase through 16 s rRNA sequencing. Maximum xylanase yield of 15.51 ± 2.08 U/mL was achieved under optimal fermentation conditions (5% inoculum, 20 g/L xylan, pH 6.0, for 24 h). After purification via ammonium sulfate precipitation and High-S ion exchange chromatography, electrophoretic purity xylanase was obtained with a 28-fold purification and specific activity of 244.97 U/mg. Xylanase had an optimal pH of 6.5 and temperature of 60 °C and displayed thermostability at 30 °C and 40 °C with 48.56% and 45.97% remaining activity after 180 min, respectively. The xylanase retained more than 82.97% of its activity after incubation for 24 h at pH 5.0 and was sensitive to metal ions, especially Mg2+ and Li+. Purified xylanase showed a molecular weight of 23 kDa on SDS-PAGE, and partial peptide sequencing revealed homology to the endo-1,4-beta-xylanase with a molecular weight of 23.3 kDa through LC/MS–MS (liquid chromatography-tandem mass spectrometry). This study suggests that the purified xylanase is easier to purify and enriches low molecular weight xylanases from bacteria source.

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Data availability

All data generated or analyzed during this study are included in this published article and its additional files. The datasets generated during the study are available in the NCBI database (https://blast.ncbi.nlm.nih.gov/Blast.cgi).

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Funding

The work was supported by Key Project of Yunnan Academy of Tobacco Sciences (2021YL04).

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Author notes

  1. Ge Zhang and Zhihao Li contributed equally to this work.

Authors and Affiliations

  1. Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, 266101, China

    Ge Zhang, Zhihao Li, Guoqiang Chen, Shuaijun Deng & Haibo Zhang

  2. Yichang Tobacco Company of Hubei Province, Yichang, 443000, China

    Zhihao Li

  3. China Tobacco Sichuan Industrial Co., Ltd, Chengdu, 610000, China

    Liang Zhang & Wen Cai

  4. Technology Center of China Tobacco Yunnan Industrial Co., Ltd, Kunming, 650032, China

    Lijun Wu

  5. Technology Center of China Tobacco Shandong Industrial Co., Ltd, Qingdao, 266101, China

    Hongtao Li

  6. Key Laboratory of Tobacco Biology and Processing, Tobacco Research Institute of Chinese Academy of Agricultural Sciences, Qingdao, 266101, China

    Haobao Liu

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Contributions

GZ and ZL contributed equally to this work. GZ and ZL carried out the experiments in this study and drafted the manuscript. HL, LW, and HL conducted the design of the experiment and helped draft and finalized the manuscript. WC, LZ, and HZ revised the manuscript. ZL and GZ assisted with the screening and fermentation experiments. GC and SD participated in the xylanase activity detection and data analysis work. All authors read and approved the final manuscript.

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Correspondence to Lijun Wu, Hongtao Li or Haobao Liu.

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Zhang, G., Li, Z., Chen, G. et al. Purification and characterization of the low molecular weight xylanase from Bacillus cereus L-1. Braz J Microbiol 54, 2951–2959 (2023). https://doi.org/10.1007/s42770-023-01129-5

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