Department of Biology, Science Faculty, Ataturk University, 25240, Erzurum, Turkey.
Department of Biology, Science Faculty, Ataturk University, 25240, Erzurum, Turkey.
Protein Expr Purif. 2021 Apr;180:105819. doi: 10.1016/j.pep.2021.105819. Epub 2021 Jan 5.
Lipase producer bacterium isolated from Erzurum was identified as Aeromonas caviae LipT51 (GenBank ID: MN818567.1) by 16S rDNA sequencing and conventional methods. Extracellular lipase was purified by ammonium sulphate precipitation, centrifugal filtration, and anion-exchange chromatography resulting in 6.1-fold purification with 28% final yield. Molecular weight was 31.6 kDa on SDS-PAGE. Lipase was stable over a broad range of pH (6-11) and temperature (25-70 °C), and showed optimum activity at pH 9 and 60 °C. Km and Vmax for pNPP hydrolysis were 0.88 mM and 34.2 U/mg protein, respectively. Ba, Ca, Co, Cu, Fe, and Mg increased activity, while Mn, Mo, Ni, Zn, and other additives partially decreased. Activity and stability increased with laundry detergent and slightly decreased with handwash and dishwashing detergents. Alkaline and thermostable lipase from newly isolated A. caviae has been shown for the first time to be remarkably compatible with laundry detergent and improve washing performance by enhanced oil-stain removal.
从埃尔祖鲁姆分离出的脂肪酶产生菌通过 16S rDNA 测序和常规方法鉴定为豚鼠气单胞菌 LipT51（GenBank ID：MN818567.1）。通过硫酸铵沉淀、离心过滤和阴离子交换层析，从细胞外粗提物中分离和纯化了脂肪酶，得到了 6.1 倍的纯化倍数，最终产率为 28%。SDS-PAGE 显示脂肪酶的分子量为 31.6 kDa。脂肪酶在广泛的 pH 值（6-11）和温度（25-70°C）范围内稳定，在 pH 9 和 60°C 时表现出最佳活性。pNPP 水解的 Km 和 Vmax 分别为 0.88 mM 和 34.2 U/mg 蛋白。Ba、Ca、Co、Cu、Fe 和 Mg 提高了酶活性，而 Mn、Mo、Ni、Zn 和其他添加剂则部分降低了酶活性。活性和稳定性随洗衣洗涤剂增加，而随手洗和洗碗洗涤剂略有降低。从新分离的豚鼠气单胞菌中首次显示出碱性和热稳定的脂肪酶与洗衣洗涤剂具有极好的相容性，并通过增强去除油斑来提高洗涤性能。
