期刊介绍 期刊简介 收录情况 栏目设置 发表周期 读者对象 联系我们 投稿须知 投稿方式 征稿范围与栏目介绍 书写要求 数据论文投稿须知 测序类论文要求 增强出版附加材料 实验伦理要求 文章的署名 关于版权 发表费及稿费 重复度检测 审稿程序及提前发表 政策制度 出版道德声明 生成性人工智能(AI)政策 开放获取声明 数据共享政策 评审政策 1河南工业大学 粮油食品学院，河南 郑州 450001;2中国农业大学 工学院，北京 100083;3中原食品实验室，河南 漯河 462000;4中国农业大学 食品科学与营养工程学院，北京 100083 魏家永：实验数据收集，数据整理，撰写文章；谭琳慧：实验数据收集；李延啸：研究课题指导，实验方法设计；王楠楠：研究材料采集；江正强：研究课题指导，方案设计，审阅与修订。 国家自然科学基金 22378419 国家自然科学基金(22378419)

1College of Food Science and Engineering, Henan University of Technology, Zhengzhou 450001, Henan, China;2College of Engineering, China Agricultural University, Beijing 100083, China;3Food Laboratory of Zhongyuan, Luohe 462000, Henan, China;4College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China the National Natural Science Foundation of China 22378419 This work was supported by the National Natural Science Foundation of China (22378419). 背景 天然芽孢杆菌产β-1,3-1,4-葡聚糖酶水平整体偏低，难以大规模应用。 目的 从土壤中筛选高产β-1,3-1,4-葡聚糖酶的菌株，研究其相关酶学性质，为开发新型β-1,3-1,4-葡聚糖酶及其在食品等工业中的应用提供技术支撑。 方法 通过形态学观察和16S rRNA基因序列分析进行菌种鉴定。采用单因素试验对菌株的液体发酵产酶条件进行优化，经硫酸铵分级沉淀和Q-Sepharose Fast Flow阴离子交换层析纯化后，测定其酶学性质。 结果 从土壤中分离获得1株高产β-1,3-1,4-葡聚糖酶的细菌，鉴定为斯皮氏芽孢杆菌( Bacillus spizizenii )，命名为HWD28。以6.0%大麦粉为碳源，4.0%干酪素为氮源，0.5%吐温-20为表面活性剂，在初始pH 8.0和40 ℃条件下发酵4 d，发酵液酶活力高达4 048 U/mL，高于目前已报道细菌产β-1,3-1,4-葡聚糖酶活力。两步纯化获得分子量约为28 kDa的电泳级纯酶BsGluA。该酶最适反应条件为pH 7.0、50 ℃，并在pH 5.0-10.0范围内保持良好的稳定性。60 ℃处理30 min后仍能保持80%以上酶活。BsGluA对β-1,3-1,4-葡聚糖底物表现出严格专一性，仅能高效水解大麦β-葡聚糖、燕麦β-葡聚糖和地衣多糖。 结论 斯皮氏芽孢杆菌HWD28产β-1,3-1,4-葡聚糖酶产量高且酶学性质好，在食品和饲料工业中具有良好的应用前景。 Background The low production of β-1,3-1,4-glucanase by naturally occurring Bacillus strains limits large-scale industrial applications. Objective To isolate a soil-derived strain producing high levels of β-1,3-1,4-glucanase and characterize the properties of the enzyme, thus providing technical support for the development of novel β-1,3-1,4-glucanases for use in the food and related industries. Methods The strain was identified by morphological observation and 16S rDNA sequence analysis. Single-factor experiments were performed to optimize liquid fermentation conditions for enzyme production. The enzyme was purified by ammonium sulfate fractional precipitation followed by Q-Sepharose Fast Flow anion-exchange chromatography, and its enzymatic properties were subsequently characterized. Results A strain with a high yield of β-1,3-1,4-glucanase was isolated from soil and identified as B . spizizenii (designated HWD28). Under optimized conditions (6.0% barley flour as the carbon source, 4.0% casein as the nitrogen source, and 0.5% Tween 20 as a surfactant), fermentation at an initial pH 8.0 and 40 ℃ for 4 days yielded an activity of 4 048 U/mL, which is higher than most previously reported bacterial β-1,3-1,4-glucanases. A two-step purification procedure yielded an electrophoretically homogeneous enzyme (BsGluA) with an apparent molecular weight of approximately 28 kDa. The optimal reaction conditions for BsGluA were pH 7.0 and 50 ℃, and the enzyme was stable within a range of pH 5.0-10.0. After incubation at 60 ℃ for 30 min, the enzyme retained 80% of the initial activity. BsGluA exhibited strict specificity toward β-1,3-1,4-glucans and efficiently hydrolyzed only barley β-glucan, oat β-glucan, and lichenan. Conclusion B . spizizenii HWD28 produces a β-1,3-1,4-glucanase with a high yield and desirable enzymatic properties, demonstrating considerable potential for application in the food and feed industries. 魏家永，谭琳慧，李延啸，王楠楠，江正强. 斯皮氏芽孢杆菌产 β-1,3-1,4- 葡聚糖酶的发酵条件优化与酶学性质[J]. 微生物学通报, 2026, 53(10): 5379-5392 收稿日期: 2026-01-02 在线发布日期: 2026-10-19