Febriani, Ihsanawat, Rukman Hertadi, Fida Madayanti, Akhmaloka
The crude extract of lipase from Thermus aquaticus isolated from Kamojang A hot spring, West Java, Indonesia was partialy purified using ammonium fractionations and hydrophobic interactions-based chromatography. The highest specific activity of the fractions collected from the chromatography was still remaining activity at about 19.46 unit/mg protein. The optimum activity of lipase at 65 °C was attained at pH 8.0. As the temperature varied at this optimum pH, if was found three temperature optima i.e. 55, 65, and 75 °C indicating that the enzyme is consisted of more than one lipases with different level of thermostability. Further analysis at each optimum temperature showed that the enzymes which are active at temperature 55 and 65 °C comprised of more than one active lipase as indicated by the appearance of more than one pH optima, while at 75 °C, it has only a single optimum pH i.e pH 8.5, thereby confirming a single active lipase exist in the condition. Further characterisation of the enzyme showed that Ca2+ and Mn2+ ions could increasing the enzyme activity but the reverse effect was exhibited by Co2+, K+, Na+, Ba2+, Cu2+, Fe3+, Sn2+, Fe3+ions. The addition of some organic molecules, such as EDTA, DTT, SDS, and PMSF were sharply reduced the enzyme activity. Investigat ion on the substrate specificity was found that the enzyme prefer to hydrolyze p-nitrophenyl myristate and palmitate with specific activity of about 9.35 and 9.77 unit/mg protein, respectively. © OmicsVista Group, All rights reserved.
Department of Chemistry, University of Syiah Kuala, Banda Aceh, Tgk Tanoh Abee 3 Darussalam, Indonesia; Biochemistry Research Group, Institut Teknologi Bandung, Bandung, Ganesha 10, Indonesia
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