Ali Imran, Akbar Ali, Yanwisetpakdee Benjawan, Prasongsuk Sehanat, Lotrakul Pongtharin, Punnapayak Hunsa
Plant Biomass Utilization Research Unit, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand ; Institute of Biochemistry, University of Balochistan, Quetta 87300, Pakistan.
Food Engineering and Bioprocess Technology, School of Environment, Resources and Development, Asian Institute of Technology, Klong Luang, Pathumthani 12120, Thailand.
Biomed Res Int. 2014;2014:106937. doi: 10.1155/2014/106937. Epub 2014 May 14.
An obligate halophilic Aspergillus gracilis which was isolated from a hypersaline man-made saltern from Thailand was screened for its potential of producing extracellular α -amylase in the previous studies. In this study the α -amylase was extracted and purified by the help of column chromatography using Sephadex G-100 column. Presence of amylase was verified by SDS-PAGE analysis, showing a single band of approximately 35 kDa. The specific activity of the enzyme was found to be 131.02 U/mg. The Lineweaver-Burk plot showed the V max and K m values of 8.36 U/mg and 6.33 mg/mL, respectively. The enzyme was found to have the best activity at 5 pH, 60°C, and 30% of NaCl concentration, showing its polyextremophilic nature. The use of various additives did not show much variation in the activity of enzyme, showing its resilience against inhibitors. The enzyme, when tested for its use for synthetic waste water remediation by comparing its activity with commercial amylase in different salt concentrations showed that the α -amylase from A. gracilis was having better performance at increasing salt concentrations than the commercial one. This shows its potential to be applied in saline waste water and other low water activity effluents for bioremediation.
在之前的研究中,从泰国一个高盐度人工盐场分离出的嗜盐纤细曲霉被筛选用于生产胞外α-淀粉酶的潜力。在本研究中,借助使用葡聚糖G-100柱的柱色谱法提取并纯化了α-淀粉酶。通过SDS-PAGE分析验证了淀粉酶的存在,显示出一条约35 kDa的单一带。发现该酶的比活性为131.02 U/mg。Lineweaver-Burk图显示Vmax和Km值分别为8.36 U/mg和6.33 mg/mL。发现该酶在pH 5、60°C和30%的NaCl浓度下具有最佳活性,表明其多极端嗜性。使用各种添加剂并未显示出酶活性有太大变化,表明其对抑制剂具有抗性。通过在不同盐浓度下将其活性与商业淀粉酶进行比较,测试该酶用于合成废水修复时发现,来自纤细曲霉的α-淀粉酶在盐浓度增加时比商业淀粉酶表现更好。这表明其有潜力应用于含盐废水和其他低水分活度废水的生物修复。