Dong Mingjie, Yang Yunjuan, Tang Xianghua, Shen Jidong, Xu Bo, Li Junjun, Wu Qian, Zhou Junpei, Ding Junmei, Han Nanyu, Mu Yuelin, Huang Zunxi
School of Life Science, Yunnan Normal University, Kunming, 650500 People's Republic of China.
Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, No.1 Yuhua District, Chenggong, Kunming, 650500 Yunnan People's Republic of China.
Springerplus. 2016 Jun 16;5(1):746. doi: 10.1186/s40064-016-2360-9. eCollection 2016.
Bos frontalis, which consumes bamboo and weeds, may have evolved unique gastrointestinal microorganisms that digest cellulase. A Paenibacillus sp. YD236 strain was isolated from B. frontalis feces, from which a GH8 endoglucanase gene, pglue8 (1107 bp, 54.5 % GC content), encoding a 368-residue polypeptide (PgluE8, 40.4 kDa) was cloned. PgluE8 efficiently hydrolyzed barley-β-d-glucan followed by CMC-Na, soluble starch, laminarin, and glucan from black yeast optimally at pH 5.5 and 50 °C, and retained 78.6, 41.6, and 34.5 % maximum activity when assayed at 20, 10, and 0 °C, respectively. Enzyme activity remained above 176.6 % after treatment with 10.0 mM β-mercaptoethanol, and was 83.0, 78, and 56 % after pre-incubation in 30 % (w/v) NaCl, 16.67 mg/mL trypsin, and 160.0 mg/mL protease K, respectively. Cys23 and Cys364 residues were critical for PgluE8 activity. pglue8, identified from B. frontalis feces for the first time in this study, is a potential alternative for applications including food processing, washing, and animal feed preparation.
以竹子和杂草为食的竹牛可能进化出了独特的能消化纤维素酶的胃肠道微生物。从竹牛粪中分离出一株芽孢杆菌属YD236菌株,从中克隆出一个GH8内切葡聚糖酶基因pglue8(1107 bp,GC含量54.5%),该基因编码一个368个残基的多肽(PgluE8,40.4 kDa)。PgluE8在pH 5.5和50℃时能高效水解大麦-β-D-葡聚糖,其次是羧甲基纤维素钠、可溶性淀粉、海带多糖和黑酵母葡聚糖,在20、10和0℃测定时分别保留最大活性的78.6%、41.6%和34.5%。用10.0 mMβ-巯基乙醇处理后酶活性保持在176.6%以上,在30%(w/v)NaCl、16.67 mg/mL胰蛋白酶和160.0 mg/mL蛋白酶K中预孵育后分别为83.0%、78%和56%。Cys23和Cys364残基对PgluE8活性至关重要。本研究首次从竹牛粪中鉴定出的pglue8,在食品加工、洗涤和动物饲料制备等应用中是一种潜在的替代品。