Department of Biotechnology, Motilal Nehru National Institute of Technology, Allahabad 211 004, Uttar Pradesh, India.
Bioresour Technol. 2012 Sep;120:300-4. doi: 10.1016/j.biortech.2012.06.018. Epub 2012 Jun 16.
The aim of present work was to bioremediate the waste cooking oil using a novel lipase produced in solid medium containing waste grease and wheat bran by Penicillium chrysogenum. Enzyme extracted with phosphate buffer was purified 10.6 and 26.28-fold after 90% ammonium sulfate precipitation and ion-exchange chromatography, respectively. The partial characterization of enzyme revealed its K(m) and V(max) value for p-nitrophenolpamitate as 0.4mM and 47.61 U/ml, respectively. The relative molecular mass of lipase was 40 kDa by SDS-PAGE and confirmed by zymogram. Purified lipase was most stable at 40°C and at 8.0 pH. Lipase activity was enhanced by metal ions such as Mg(2+), Fe(2+), Ca(2+) and non-ionic surfactant TritonX-100, while suppressed in the presence of SDS. Crude lipase was applied on cooking oil waste and the acid value was 26.92 mg/g. This showed that the enzyme could be employed for the bioremediation of used cooking oil.
本工作旨在利用新型脂肪酶从含废油脂和麦麸的固体培养基中生物修复废烹饪油。用磷酸盐缓冲液提取的酶经 90%硫酸铵沉淀和离子交换层析分别纯化 10.6 和 26.28 倍。酶的部分特性研究表明,其对 p-硝基苯棕榈酸酯的 K(m)和 V(max)值分别为 0.4mM 和 47.61 U/ml。SDS-PAGE 显示脂肪酶的相对分子质量为 40 kDa,通过酶谱得到证实。纯化的脂肪酶在 40°C 和 8.0 pH 下最稳定。脂肪酶活性被 Mg(2+)、Fe(2+)、Ca(2+)等金属离子和非离子表面活性剂 TritonX-100 增强,而在 SDS 存在下受到抑制。粗脂肪酶应用于废烹饪油,酸值为 26.92mg/g。这表明该酶可用于生物修复废烹饪油。