芽孢杆菌属HMM所产纤维素酶的纯化与特性分析
Purification and characterization of cellulase produced by Bacillus sp. HMM.
作者信息
Huang Lanqian, Mo Lei, Jiang Jiaming, Huo Yu, Yu Shisong, Ou Jianmei, Liu Siyu, Xie Xinyu, Yan Guyu, Liao Chuzun, Wu Shangxin, Zhang Huiqing, Huang Shiru, Wang Qing, Shang Changhua
机构信息
Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection (Guangxi Normal University), Ministry of Education & Guangxi Key Laboratory of Landscape Resources Conservation and Sustainable Utilization in Lijiang River Basin (Guangxi Normal University) & University Engineering Research Center of Bioinformation and Genetic Improvement of Specialty Crops, Guilin, 541006, Guangxi, China.
出版信息
Antonie Van Leeuwenhoek. 2025 Jun 27;118(8):97. doi: 10.1007/s10482-025-02112-w.
Cellulose is long-chain glucose polymer, which is linked by β-1,4-glycosidic bond. Its degradation relies on the synergistic action of multi-component cellulase system, including endoglucanase, exoglucanase and β-glucosidase. Cellulase hydrolyzes cellulose to produce glucose through the sequential reactions. While the biodegradation of lignocellulosic wastes has been extensively studied, researches on cellulase production with the mixed agricultural wastes as carbon sources remain limited. To address this gap, this study innovatively employed wheat bran (WB), sugarcane bagasse (SB) and rice hull (RH) to prepare the mixed carbon sources for cellulase production by Bacillus sp. HMM. Bacillus sp. HMM in culture medium with WB showed the highest cellulase activity (3.93 ± 0.13 U/mL) and reducing sugar content (0.97 ± 0.02 mg/mL) at 12 h. Cellulase was purified by (NH)SO and DEAE-52 column, and investigated by SDS-PAGE. Mw of purified cellulase was between 50 and 70 kDa with purification fold of 23.10. This cellulase had endoglucanase, exoglucanase and β-glucosidase activities, and the wide substrate specificity, which was conducive to the complete degradation of lignocellulose. CMCase activities showed the high resistance under stress conditions such as metal ions, surfactants and inhibitors. CMCase activities still retained over 90% under the treatment of most additives for 4 h, especially DTT greatly promoted CMCase activity, reaching 129.2% of control. Bacillus sp. HMM could efficiently utilize wheat bran to produce cellulase for the conversion of lignocellulosic biomass.
纤维素是一种长链葡萄糖聚合物,由β-1,4-糖苷键连接。其降解依赖于多组分纤维素酶系统的协同作用,该系统包括内切葡聚糖酶、外切葡聚糖酶和β-葡萄糖苷酶。纤维素酶通过一系列反应将纤维素水解产生葡萄糖。虽然木质纤维素废弃物的生物降解已得到广泛研究,但以混合农业废弃物为碳源生产纤维素酶的研究仍然有限。为了填补这一空白,本研究创新性地采用麦麸(WB)、甘蔗渣(SB)和稻壳(RH)制备混合碳源,用于芽孢杆菌HMM生产纤维素酶。在含有WB的培养基中,芽孢杆菌HMM在12小时时显示出最高的纤维素酶活性(3.93±0.13 U/mL)和还原糖含量(0.97±0.02 mg/mL)。纤维素酶通过硫酸铵和DEAE-52柱进行纯化,并通过SDS-PAGE进行研究。纯化后的纤维素酶分子量在50至70 kDa之间,纯化倍数为23.10。这种纤维素酶具有内切葡聚糖酶、外切葡聚糖酶和β-葡萄糖苷酶活性,且底物特异性广泛,有利于木质纤维素的完全降解。羧甲基纤维素酶(CMCase)活性在金属离子、表面活性剂和抑制剂等应激条件下表现出高抗性。在大多数添加剂处理4小时后,CMCase活性仍保留超过90%,尤其是二硫苏糖醇(DTT)极大地促进了CMCase活性,达到对照的129.2%。芽孢杆菌HMM能够有效地利用麦麸生产纤维素酶,用于木质纤维素生物质的转化。