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A Novel Chitosanase from Penicillium oxalicum M2 for Chitooligosaccharide Production: Purification, Identification and Characterization

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Abstract

This study discovered a novel chitosanase from Penicillium oxalicum M2 based on a new screening strategy. An extracellular chitosanase was isolated and purified from the fermentation broth of Penicillium oxalicum M2. A 19.34-fold purification was achieved on a cation exchange column. Using sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, chitosanase was determined at approximately 42 kDa without any subunits. The sequence of peptide in the protein was identified as SALNKNYITNFSTLR by MALTI-TOF/TOF MS. The maximum catalytic activity of the purified enzyme was 60.45 U/mg at the optimum pH and temperature of 5.5 and 60 °C. The enzyme activity held stability in the range of 35–50 °C and pH 3–4.5. Ca2+, Mn2+, non-ionic surfactants (Tween 20/40/60/80 and Trition X-100) and some common reducing agents (DTT and β-ME) could significantly activate chitosanase. The purified enzyme showed rigorous specificity to chitosan as a substrate. The hydrolysate in the final stage of hydrolysis consisted of chitooligosaccharides with a degree of polymerization ranging from 2 to 5 and without glucosamine or acetylglucosamine. The monomeric enzyme obtained by one-step purification reveal applications potential in sugar industry, and expanded our understanding of the GH75 family chitosanases simultaneously.

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Acknowledgements

This work is supported by National Key R&D Program of China (2018YFD0901104), National Natural Science Foundation of China (31800679), National Natural Science Foundation of China (31801575), China Postdoctoral Science Foundation (2018M642167), Fundamental Research Funds for the Central Universities Institute of Food Science and Technology International Journal of Food Science & Technology (JUSRP121008) and China Agriculture Research System of MOF and MARA

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, 214122, China

    Shining Cao, Pei Gao, Wenshui **a & Bin Wang

  2. Synergetic Innovation Center of Food Safety and Quality Control, Jiangsu Province, Wuxi, 214122, China

    Shining Cao, Pei Gao, Wenshui **a & Bin Wang

  3. Department of Food Science and Technology, National University of Singapore, Science Drive 2, Singapore, 117546, Singapore

    Shaoquan Liu

  4. National University of Singapore (Suzhou) Research Institute, No. 377 Linquan Street, Suzhou Industrial Park, Suzhou, 215123, Jiangsu, China

    Shaoquan Liu

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  1. Shining Cao
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Correspondence to Wenshui **a.

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Cao, S., Gao, P., **a, W. et al. A Novel Chitosanase from Penicillium oxalicum M2 for Chitooligosaccharide Production: Purification, Identification and Characterization. Mol Biotechnol 64, 947–957 (2022). https://doi.org/10.1007/s12033-022-00461-9

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