Abstract
An extracellular lipase from Aureobasidium pullulans was obtained and purified with a specific activity of 17.7 U/mg of protein using ultrafiltration and a DEAE-Sepharose Fast Flow column. Characterization of the lipase indicated that it is a novel finding from the species A. pullulans. The molecular weight of the lipase was 39.5 kDa, determined by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme exhibited its optimum activity at 40 °C and pH of 7. It also showed a remarkable stability in some organic solutions (30%, v/v) including n-propanol, isopropanol, dimethyl sulfoxide (DMSO), and hexane. The catalytic activity of the lipase was enhanced by Ca2+ and was slightly inhibited by Mn2+ and Zn2+ at a concentration of 10 mmol/L. The lipase was activated by the anionic surfactant SDS and the non-ionic surfactants Tween 20, Tween 80, and Triton X-100, but it was drastically inhibited by the cationic surfactant cetyl trimethyl ammonium bromide (CTAB). Furthermore, the lipase was able to hydrolyze a wide variety of edible oils, such as peanut oil, corn oil, sunflower seed oil, sesame oil, and olive oil. Our study indicated that the lipase we obtained is a potential biocatalyst for industrial use.
概 要
目 的
从出芽短梗霉所产的脂肪酶中筛选具有独特酶学 性质的脂肪酶
创新点
发现了一种新的产自出芽短梗霉的脂肪酶, 并对 其酶学性质进行了研究。
方 法
通过超滤和 DEAE-Sepharose Fast Flow 阴离子层 析柱方法对脂肪酶进行纯化, 随后分别用对硝基 酚邻酸盐(pNPP) 法对纯化得到的脂肪酶进行了 酶学性质研究, 并用酸碱中和法检测了脂肪酶对 可食用油脂的水解。
结 论
对分离纯化得到的脂肪酶的酶学性质研究表明, 该酶的分子量为39.5 kDa, 具有一个亚基, 为胞外酶。最佳催化温度为40 °C, 最佳催化pH 为7。该酶对一些有机溶剂、表面活性剂和离子具有优良的抗性。此外, 它可以水解常见的食用油。这些良好的特性使该脂肪酶有可能被应用于洗涤剂生产、生物柴油合成和食品制造等一些工业领域。
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Li, Y., Liu, Tj., Zhao, Mj. et al. Screening, purification, and characterization of an extracellular lipase from Aureobasidium pullulans isolated from stuffed buns steamers. J. Zhejiang Univ. Sci. B 20, 332–342 (2019). https://doi.org/10.1631/jzus.B1800213
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DOI: https://doi.org/10.1631/jzus.B1800213