从尼泊尔地热泉分离出的BTPS-2中纯化并鉴定一种优质耐热性藻类淀粉液化α-淀粉酶
Purification and characterization of a noble thermostable algal starch liquefying alpha-amylase from BTPS-2 isolated from geothermal spring of Nepal.
作者信息
Timilsina Parash Mani, Pandey Gyanu Raj, Shrestha Asmita, Ojha Manish, Karki Tika Bahadur
机构信息
Department of Biotechnology, Kathmandu University, Dhulikhel, 6250, Nepal.
Biotechnological Research and Developmental Center, Bharatpur, Chitwan, 44200, Nepal.
出版信息
Biotechnol Rep (Amst). 2020 Nov 1;28:e00551. doi: 10.1016/j.btre.2020.e00551. eCollection 2020 Dec.
A thermophilic strain, BTPS-2 was isolated from Bhurung geothermal spring of Nepal. The 16 s rRNA sequence showed 99.8 % similarity with the type strain DSM 3670. The morphological, physiological and biochemical properties were similar to the type strain. Alpha-amylase from BTPS-2 was purified to 19-fold purification by DEAE-Cellulose ion exchange chromatography. The K value of amylase on starch was 0.51 ± 0.05 mg/mL. The optimum pH and temperature were 7.0 and 70 °C. SDS-PAGE analysis showed a single band at 100 kDa. The half-life of the enzyme at 80 °C was 2.81 h. The enzyme showed an inhibitory effect in the presence of Fe, Pb, Sn and Hg at 10 mM concentrations. TLC analysis showed that the enzyme is a liquifying alpha-amylase. The enzyme reduced the viscosity of algal biomass suspension up to 74.2 ± 0.17 % which was more efficient than alpha-amylase (80.5 ± 0.2 %).
从尼泊尔布隆地热泉中分离出嗜热菌株BTPS - 2。其16 s rRNA序列与模式菌株DSM 3670显示出99.8%的相似性。形态、生理和生化特性与模式菌株相似。通过DEAE -纤维素离子交换色谱法将来自BTPS - 2的α -淀粉酶纯化至19倍。淀粉酶对淀粉的K值为0.51±0.05 mg/mL。最适pH和温度分别为7.0和70°C。SDS - PAGE分析在100 kDa处显示单一条带。该酶在80°C下的半衰期为2.81 h。在10 mM浓度的铁、铅、锡和汞存在下，该酶表现出抑制作用。TLC分析表明该酶是一种液化α -淀粉酶。该酶将藻类生物质悬浮液的粘度降低了74.2±0.17%，比α -淀粉酶（80.5±0.2%）更有效。
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