Wang Xiaobei, Cheng Huaixu, Lu Mingsheng, Fang Yaowei, Jiao Yuliang, Li Weijuan, Zhao Gengmao, Wang Shujun
a Marine Resources Development Institute of Jiangsu , Lianyungang , PR China.
b Key Laboratory of Marine Biology , Nanjing Agricultural University , Nanjing , Jiangsu , PR China.
Biofouling. 2016 Nov;32(10):1223-1233. doi: 10.1080/08927014.2016.1239722.
Dental plaque is a biofilm of water-soluble and water-insoluble polysaccharides, produced primarily by Streptococcus mutans. Dextranase can inhibit biofilm formation. Here, a dextranase gene from the marine microorganism Arthrobacter oxydans KQ11-1 is described, and cloned and expressed using E. coli DH5α competent cells. The recombinant enzyme was then purified and its properties were characterized. The optimal temperature and pH were determined to be 60°C and 6.5, respectively. High-performance liquid chromatography data show that the final hydrolysis products were glucose, maltose, maltotriose, and maltotetraose. Thus, dextranase can inhibit the adhesive ability of S. mutans. The minimum biofilm inhibition and reduction concentrations (MBIC and MBRC) of dextranase were 2 U ml and 5 U ml, respectively. Scanning electron microscopy and confocal laser scanning microscope (CLSM) observations confirmed that dextranase inhibited biofilm formation and removed previously formed biofilms.
牙菌斑是一种主要由变形链球菌产生的水溶性和水不溶性多糖生物膜。葡聚糖酶可抑制生物膜形成。在此,描述了一种来自海洋微生物氧化节杆菌KQ11 - 1的葡聚糖酶基因,并使用大肠杆菌DH5α感受态细胞进行克隆和表达。然后对重组酶进行纯化并表征其性质。确定最佳温度和pH分别为60°C和6.5。高效液相色谱数据表明,最终水解产物为葡萄糖、麦芽糖、麦芽三糖和麦芽四糖。因此,葡聚糖酶可抑制变形链球菌的黏附能力。葡聚糖酶的最小生物膜抑制和减少浓度(MBIC和MBRC)分别为2 U/ml和5 U/ml。扫描电子显微镜和共聚焦激光扫描显微镜(CLSM)观察证实,葡聚糖酶抑制生物膜形成并去除先前形成的生物膜。