eng Oriental Scientific Publishing Company Biosciences Biotechnology Research Asia 0973-1245 2017-09-25 14 3 1081 1088 10.13005/bbra/2545 27846 article Anita H. Permana 1,4 Fida Madayanti Warganegara 1 Deana Wahyuningrum 2 Made Puspasari Widhiastuty 1 Akhmaloka 1,3 Biochemistry Research Group, Faculty of Mathematics and Natural Science, Institut Teknologi Bandung, Indonesia. Organic Chemistry Research Group, Faculty of Mathematics and Natural Science, Institut Teknologi Bandung, Indonesia. Department of Chemistry, Faculty of Science and Computer, Universitas Pertamina, Indonesia. Study Program of Quality Assurance of Food Industry, Politeknik AKA Bogor, Indonesia. Heterologous expression and purification of thermostable lipase from Geobacillus thermoleovorans PPD2 had been carried out through Escherichia coli BL21 as host. Two bands obtained showed lipolytic activity with the size at around 51 (LipA) and 43 (LipB) kDa, respectively. The activities were identified by zymogram analysis, while the control protein from Escherichia coli BL21(DE3) do not show any lipolytic activity. Purification of crude extract using chromatography affinity Ni-NTA resulted one dominant band of LipA, meanwhile LipB did not appeared on the gel. Another purification for LipB was carried out by acetone fractionation. Both of LipA and LipB showed high activity toward medium chain length substrates, with optimum activity at 50^oC and pH 8.5. The activities of LipA and LipB showed tolerance toward short chain alcohols, such as methanol, ethanol, n-propanol, and isopropanol. https://www.biotech-asia.org/vol14no3/heterologous-expression-and-characterization-of-thermostable-lipases-from-geobacillus-thermoleovorans-ppd2-through-escherichia-coli/ Acetone Fractionation Geobacillus Thermoleovorans PPD2; Thermostable Lipase; Recombinant Protein; Ni-NTA Affinity Chromatography;