Isolation and characterization of lipase enzyme from plant seed

Lipases in the group of hydrolases are called enzymes that provide hydrolysis of triglycerides in aqueous media. In this study, it was aimed to isolate and characterize lipase enzyme from the broom seed (Sorghum vulgare). Broom seed lipase was purified 2-fold by using Triple Phase System and the enzyme activity was determined from the broom seed by titrimetric method. 7 different substrates were used to determine the substrate specificity of broom seed lipase. Olive oil was determined as the best substrate for the broom seed lipase. In the kinetic studies, Michaelis–Menten values for tributyrin were found as Km: 19,28 mM and Vmax: 45,87 U/min.mg.enzyme. The optimum reaction time of the lipase obtained from the broom seed was determined as 6 minutes. The purified enzyme had the maximum activity at pH = 6 and 60 ºC. In the determination of the storage stability, the loss of the activity of the broom seed lipase waiting at -20 °C for 1 year was determined as 25,6%. It was determined that CaCl2 acted as activator in enzyme activity and the increase in enzyme activity at 2 mM CaCl2 concentration was determined as 121%. The calculated activation energy of broom seed lipase was 11,34 kJ / mol. In addition, the effect of some metals and EDTA on enzyme activity was investigated and the activity of enzyme was increased approximately 4-fold with EDTA and Cu2+, whereas Fe2+ increased the enzyme activity approximately 2-fold. Mg2+ and Co2+ increased activity in the range of 6-10%. However, the other tested metals inhibited the enzyme in the range of 10% to 40%. The effects of detergents on the activity of broom seed lipase enzyme were also investigated. According to the results, the sodium deoxycholate used as anionic detergent did not cause any change in enzyme activity. However, the cationic detergent CTAB caused 60% increase in enzyme activity. The non-ionic Triton X-100 increased activity by 2 times while Tween-80 increased by 50%. The enzyme activity was not inhibited in any of the examples of detergent used.