Studies on the catalytic behavior of a membrane-bound lipolytic enzyme from the microalgae Nannochloropsis oceanica CCMP1779

Abstract

The catalytic behavior of a membrane-bound lipolytic enzyme (MBL-Enzyme) from the microalgae Nannochloropsis oceanica CCMP1779 was investigated. The biocatalyst showed maximum activity at 50 °C and pH 7.0, and was stable at pH 7.0 and temperatures from 40 to 60 °C. Half-lives at 60 °C, 70 °C and 80 °C were found 866.38, 150.67 and 85.57 min respectively. Thermal deactivation energy was 68.87 kJ mol^-1. The enzyme's enthalpy (ΔΗ*), entropy (ΔS*) and Gibb's free energy (ΔG*) were in the range of 65.86-66.27 kJ mol^-1, 132.38-140.64 J mol^-1 K^-1 and 107.80-115.81 kJ mol^-1, respectively. Among p-nitrophenyl esters of fatty acids tested, MBL-Enzyme exhibited the highest hydrolytic activity against p-nitrophenyl palmitate (pNPP). The K_m and V_max values were found 0.051 mM and of 0.054 mmole pNP mg protein^-1 min^-1, respectively with pNPP as substrate. The presence of Mn²⁺ increased lipolytic activity by 68.25%, while Fe³⁺ and Cu²⁺ ions had the strongest inhibitory effect. MBL-Enzyme was stable in the presence of water miscible (66% of the initial activity in ethanol) and water immiscible (71% of the initial activity in n-octane) solvents. Myristic acid was found to be the most efficient acyl donor in esterification reactions with ethanol. Methanol was the best acyl acceptor among the primary alcohols tested.

Keywords: Biochemical properties; Esterification activity; Hydrolytic activity; Membrane-bound lipolytic enzyme (MBL-Enzyme); Nannochloropsis oceanica CCMP1779.