Novel thermostable lipase produced by a thermo-halophilic bacterium that catalyses hydrolytic and transesterification reactions

Abstract

Lipase belongs to the class of hydrolytic enzymes that are widely used in the biotechnology industries. The goal of this research was to purify and characterize lipase produced from a thermo-halophilic bacterium, namely Pria Laot Sabang 80 (PLS 80). Purification was performed using ammonium sulphate fractionation, followed by gel filtration chromatography using Sepharose Cl-6B. After purification, the enzyme had a specific activity of 326.6 U/mg with a purity of 6.02 higher than the crude extract; with a molecular weight of around 50 kDa. The optimum activity was observed at 70 °C and pH 9. The activity increased in the presence of 10 mM Mn 2+ , K + and Ca 2+ ions, while Hg 2+ only slightly increased the enzyme activity. In contrast, the activity decreased in 10 mM Mg 2+ , Zn 2+ , Co 2+ , EDTA, and PMSF. The enzyme showed good hydrolytic activity on long fatty acids substrates ( p -nitrophenyl palmitate) with a value of 35.5 U/mL. It was also able to catalyze a transesterification reaction. GC-MS result showed that the biodiesel consisted of methyl octanoate (5.3%), methyl caprate (12.4%), methyl laurate (34.1%), methyl myristate (10.7%), methyl palmitate (3.9%), and methyl stearate (1.2%) when using coconut oil as the substrate. The results suggested that the lipase from PLS 80 had unique attributes that could be useful in various industrial applications.

Utilization of vanillin to prepare sulfated Calix[4]resorcinarene as efficient organocatalyst for biodiesel production based on methylation of palmitic acid and oleic acid

“…Compared to the methylation of free fatty acids, the transesterification reaction is more complicated due to the stepwise conversion mechanism of triglycerides to glycerol and methyl esters [ 11 ]. Furthermore, the nature of plant oils depends on the geochemical and climate factors; thus, it is not easy to control and adjust as a standard comparison for literature purposes [ [12] , [13] , [14] , [15] ]. In contrast, methylation of free fatty acids is more preferable due to its exact physicochemical properties; thus, a faster and more efficient production process could be comprehensively studied.…”

“…Couples of organocatalysts for the methylation of palmitic acid and oleic acid have been reported [ 14 , [29] , [30] , [31] , [32] , [33] , [34] , [35] , 37 , [41] , [42] , [43] , [44] , [45] ]. Table 1 , Table 2 showed that organocatalysts exhibit higher catalytic activity to produce methyl palmitate and methyl oleate in up to 98.3 and 97.6% yield, respectively.…”

Synthesis of neryl acetate by free lipase-catalyzed transesterification in organic solvents and its kinetics

“…Some carboxylic acids are difficult to dissolve or extremely unstable in organic solvents, whereas esters can dissolve and remain stable in almost all organic solvents (Yue et al, 2022). Therefore, transesterification of esters is more important than direct esterification of alcohol and acid, which makes enzyme catalyzed transesterification reactions to synthesize perfume ester an important research direction (Shah et al, 2004;Gómez et al, 2020;Febriani et al, 2020). The preparation of neryl acetate by lipase-catalyzed transesterification has obvious environmental advantages.…”