Journal of Pure and Applied MicrobiologyVol. 11 No. 1

Heterologous Expression of Gene Encoded Thermostable Lipase and Lipolytic Activity

Nurhasanah1,2, Santi Nurbaiti1, Fida Madayanti1 and Akhmaloka1,3

1Biochemistry Research Group, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Indonesia. 2Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Negeri Lampung, Indonesia. 3Department of Chemistry, Faculty of Sciences and Computer, Universitas Pertamina, Indonesia.

Received on 01 December 2016 and accepted on 20 January 2017

 

ABSTRACT

Three lipase genes namely LK2, LK3 and LK5 were successfully sub-cloned into expression vector pET30a. The recombinant vectors were heterologically expressed into Escherichia coli BL21 (DE3) by 1 mM IPTG induced at 37 C. The protein with the size of 32, 31 and 28 kDa with correspond to LK2, LK3 and LK5 clones were over expressed following SDS-PAGE analysis. Further analysis to quantity the level of expression showed that the protein were over expressed at around 30, 44 and 21 % of the total protein for LK2, LK3 and LK5 respectively. The lipolytic activity of the proteins by using lauric acid (C12) as substrate at 50C, appeared that LK2 showed the highest activity among the other (1.49U/mg) followed by LK5 (1,10U/mg) and the lowest activity is LK3 (0.94U/mg).

Keywords : Thermostable lipase, Heterologous expression, Lipolytic activity.