Panosyan Hovik, Di Donato Paola, Poli Annarita, Nicolaus Barbara
Department of Biochemistry, Microbiology and Biotechnology, Yerevan State University, Alex Manoogian 1, 0025, Yerevan, Armenia.
Institute of Biomolecular Chemistry, National Council of Research (C.N.R.), Via Campi Flegrei 34, 80078, Pozzuoli, NA, Italy.
Extremophiles. 2018 Sep;22(5):725-737. doi: 10.1007/s00792-018-1032-9. Epub 2018 May 19.
The thermal ecosystems, including geothermal springs, are proving to be source of thermophiles able to produce extracellular polysaccharides (EPSs). Among the sixteen thermophilic bacilli isolated from sediment sampled from Arzakan geothermal spring, Armenia, two best EPSs producer strains were identified based on 16S rRNA gene sequence analysis and phenotypic characteristics, and designated as Geobacillus thermodenitrificans ArzA-6 and Geobacillus toebii ArzA-8 strains. EPSs production was investigated under different time, temperature and culture media's composition. The highest specific EPSs production yield (0.27 g g dry cells and 0.22 g g dry cells for strains G. thermodenitrificans ArzA-6 and G. toebii ArzA-8, respectively) was observed after 24 h when fructose was used as sole carbon source at 65 °C and pH 7.0. Purified EPSs displayed a high molecular mass: 5 × 10 Da for G. thermodenitrificans ArzA-6 and 6 × 10 Da for G. toebii ArzA-8. Chemical composition and structure of the biopolymers, determined by GC-MS, HPAE-PAD and NMR, showed that both the two EPSs are heteropolymers composed by mannose as major monomer unit. Optical rotation values [α] of the two EPSs (2 mg ml HO) were - 142,135 and - 128,645 for G. thermodenitrificans ArzA-6 and G. toebii ArzA-8, respectively.
包括地热泉在内的热生态系统正被证明是能够产生胞外多糖(EPSs)的嗜热菌的来源。从亚美尼亚阿尔扎坎地热泉采集的沉积物中分离出的16株嗜热芽孢杆菌中,基于16S rRNA基因序列分析和表型特征鉴定出两株最佳EPSs产生菌株,分别命名为嗜热脱氮地芽孢杆菌ArzA-6和托氏地芽孢杆菌ArzA-8菌株。在不同时间、温度和培养基组成条件下研究了EPSs的产生情况。当在65℃和pH 7.0条件下使用果糖作为唯一碳源时,24小时后观察到最高的特定EPSs产量(嗜热脱氮地芽孢杆菌ArzA-6菌株为0.27 g g干细胞,托氏地芽孢杆菌ArzA-8菌株为0.22 g g干细胞)。纯化后的EPSs显示出高分子量:嗜热脱氮地芽孢杆菌ArzA-6为5×10 Da,托氏地芽孢杆菌ArzA-8为6×10 Da。通过气相色谱-质谱联用(GC-MS)、高效阴离子交换色谱-脉冲安培检测(HPAE-PAD)和核磁共振(NMR)测定的生物聚合物的化学组成和结构表明,这两种EPSs都是以甘露糖为主要单体单元组成的杂聚物。两种EPSs(2 mg ml H₂O)的旋光度[α]值,嗜热脱氮地芽孢杆菌ArzA-6为-142,135,托氏地芽孢杆菌ArzA-8为-128,645。