Recombinant Production and Analysis of Olive (Olea europae L.) Lipase Enzyme

Yurtsever M., Öztürk S. P., Vural Korkut Ş.

10th International Molecular Biology and Biotechnology Congress, Ankara, Turkey, 4 - 08 October 2021, pp.29

  • Publication Type: Conference Paper / Summary Text
  • City: Ankara
  • Country: Turkey
  • Page Numbers: pp.29
  • Yıldız Technical University Affiliated: Yes


Lipases are catalytic and versatile enzymes found in plants, animals and microorganisms. These enzymes provide the conversion of triacylglycerols to glycerol and free fatty acids. They also catalyze reactions such as esterification and transesterification under certain conditions. Due to these versatile properties, lipases are used in many areas such as the food industry, pharmaceutical and biofuel production. Although mostly microbial lipases are used in these areas, plant lipases are much less studied. Lipases are commonly found in oilseeds, leaves, rubber, and latex of plants. The full-length cDNA of olive leaf lipase was obtained in an earlier study in our laboratory (GeneBank:MG663218.2). In the present study, this cDNA was cloned with primers including KpnI and NotI recognition sites and transferred into the pGAPZαA vector. The recombinant plasmid amplified in E.coli DH5α cells was linearized and transformed into Pichia pastoris X33 competent cells and integrated into the yeast genome. Expression was carried out at 28C for 96 hours. The activity analyses of the enzyme purified with Ni-NTA resin using the His tail were performed at different reaction temperatures and times using the pNPP substrate. For X33 control group, the activity was calculated as 182.4 U/mg in the 1-hour reaction at 37C, while it was 323.6 U/mg for the olive lipase. Our tests showed olive lipase active in the range of pH 8-9.