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Expression, enzymatic properties, and product functions of a chitosanase from Bacillus halotolerans
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Ling OU, Hongyan ZHANG, Yuling WEI, Taohong WANG, Ying HUANG, Jiayan ZHANG, Naikun SHEN*
Acta Microbiologica Sinica | 2026, 66(10) : 5237 - 5254
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Acta Microbiologica Sinica | 2026, 66(10): 5237-5254
• Research Article •
Expression, enzymatic properties, and product functions of a chitosanase from Bacillus halotolerans
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Ling OU, Hongyan ZHANG, Yuling WEI, Taohong WANG, Ying HUANG, Jiayan ZHANG, Naikun SHEN*
Affiliations
  • Guangxi Key Laboratory for Polysaccharide Materials and Modification, School of Marine Sciences and Biotechnology, Guangxi Minzu University, Nanning, Guangxi, China
Published: 2026-10-04 doi: 10.13343/j.cnki.wsxb.20260245
Outline
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[Objective] To achieve efficient expression and comprehensive functional characterization of chitosanase (Csn, EC 3.2.1.132). [Methods] The chitosanase gene (CsnBh) was cloned from Bacillus halotolerans GXK615 and heterologously expressed in Escherichia coli BL21(DE3). [Results] Following purification via nickel-affinity chromatography, the recombinant CsnBh exhibited a specific activity of 400.47 U/mg, which represented 3.03-fold purification. Enzymatic characterization revealed that CsnBh demonstrated high specificity toward colloidal chitosan, with optimal activity observed at 40 ℃ and pH 6.0. The enzyme demonstrated remarkable thermal stability across a broad temperature range (30-70 ℃) and pH stability over pH 5.0-9.0. Among the metal ions tested, Mn2+ significantly enhanced enzymatic activity, whereas Cu2+, Fe2+, and Fe3+ completely inhibited the activity. Hydrolysis pattern analysis confirmed an endo-acting cleavage mechanism, with chitotriose to chitopentaose identified as the predominant oligosaccharide products. Further investigation showed that the chitooligosaccharide (COS) mixture generated by CsnBh-mediated hydrolysis exerted potent inhibitory effects against E. coli, Staphylococcus aureus, and Phytophthora nicotianae. Moreover, COS treatment significantly upregulated the activities of key plant defense-related enzymes, including superoxide dismutase, peroxidase, and phenylalanine ammonia-lyase, in tobacco leaves. [Conclusion] This study establishes both a robust theoretical foundation and a highly promising enzyme candidate for the efficient production of chitosanases and the green synthesis and application of bioactive COSs.

chitosanase  /  Bacillus halotolerans  /  heterologous expression  /  enzymatic property  /  product function
欧玲, 张红岩, 韦玉玲, 王韬闳, 黄莹, 张佳艳, 申乃坤. 耐受盐芽孢杆菌壳聚糖酶的表达、酶学性质及其产物功能分析. 微生物学报, 2026 , 66 (10) : 5237 -5254 . DOI: 10.13343/j.cnki.wsxb.20260245
Ling OU, Hongyan ZHANG, Yuling WEI, Taohong WANG, Ying HUANG, Jiayan ZHANG, Naikun SHEN. Expression, enzymatic properties, and product functions of a chitosanase from Bacillus halotolerans[J]. Acta Microbiologica Sinica, 2026 , 66 (10) : 5237 -5254 . DOI: 10.13343/j.cnki.wsxb.20260245
Funding
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  • National Natural Science Foundation of China(32560023)
  • Guangxi Zhuang Autonomous Region “Jianfeng” Action Plan(桂科JF2504850012)
  • Provincial College Student Innovation and Entrepreneurship Project of Guangxi Minzu University(202310608017)
  • Provincial College Student Innovation and Entrepreneurship Project of Guangxi Minzu University(S202510608309)
Appendix
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Year 2026 volume 66 Issue 10
166
80
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Article Info
doi: 10.13343/j.cnki.wsxb.20260245
  • Receive Date：2026-03-25
  • Online Date：2026-10-13
  • Published：2026-10-04
Article Data
Affiliations
History
  • Received：2026-03-25
  • Accepted：2026-04-17
Funding
National Natural Science Foundation of China(32560023)
Guangxi Zhuang Autonomous Region “Jianfeng” Action Plan(桂科JF2504850012)
Provincial College Student Innovation and Entrepreneurship Project of Guangxi Minzu University(202310608017)
Provincial College Student Innovation and Entrepreneurship Project of Guangxi Minzu University(S202510608309)
Affiliations
    Guangxi Key Laboratory for Polysaccharide Materials and Modification, School of Marine Sciences and Biotechnology, Guangxi Minzu University, Nanning, Guangxi, China

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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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