Production and characterization of esterase and lipase from Haloarcula marismortui

Abstract

The present study was conducted to investigate the capability of Haloarcula marismortui to synthesize esterases and lipases, and the effect of physicochemical conditions on the growth and the production of esterases and lipases. Finally, the effect of NaCl concentration and temperature on esterase and lipase activities was studied using intracellular crude extracts. In order to confirm the genomic prediction about the esterase and lipase synthesis, H. marismortui was cultured on a rich medium and the crude extracts (intra- or extracellular) obtained were assayed for both activities using p-nitrophenyl esters and triacylglycerides as substrates. Studies on the kinetics of growth and production of esterase and lipase of H. marismortui were performed, reaching a maximum growth rate of 0.053 h(-1) and maximal productions of intracellular esterase and lipase of 2.094 and 0.722 U l(-1) using p-nitrophenyl valerate and p-nitrophenyl laurate, respectively. Both enzymes were produced as growth-associated metabolites. The effects of temperature, pH, and NaCl concentration on the growth rate and production of enzymes were studied by using a Box-Behnken response surface design. The three response variables were significantly influenced by the physicochemical factors and an interaction effect between temperature and NaCl concentration was also evidenced. The surface response method estimated the following maximal values for growth rate and productions of esterase and lipase: 0.086 h(-1) (at 42.5 degrees C, pH 7.4, and 3.6 mol l(-1) NaCl), 2.3 U l(-1) (at 50 degrees C, pH 7.5, and 4.3 mol l(-1) NaCl), and 0.58 U l(-1) (at 50 degrees C, pH 7.6, and 4.5 mol l(-1) NaCl), respectively. Esterases were active at different salt concentrations, showing two optimal activities (at 0.5 and 5 mol l(-1) NaCl), which suggested the presence of two different esterases. Interestingly, in the absence of salt, esterase retained 50% residual activity. Esterases and lipase activities were maximal at 45 degrees C and inactive at 75 degrees C. This study represents the first report evidencing the synthesis of esterase and lipase by H. marismortui.

Archaeal Diversity and Their Biotechnological Potential

“…For instance, Haloarcula marismortui, a halophilic archaeon whose genome was sequenced contains genes encoding for putative esterase and lipase. These genomic predictions have been verified recently (Camacho et al 2009). They reported that Haloarcula marismortui displays esterase and lipase activity intracellularly and extracellularly.…”

“…They reported that Haloarcula marismortui displays esterase and lipase activity intracellularly and extracellularly. While lipase was accumulated mainly extracellular, esterase was generally accommodated intracellular (Camacho et al, 2009). The esterase and lipase genes of Pyrococcus furiosus, a hyperthermophilic archaeon, have been cloned in E. coli and functional properties have been determined.…”

Enzymatic activity of a novel halotolerant lipase from Haloarcula hispanica 2TK2

“…4C). Results indicated that Haloarcula hispanica 2TK2 lipase was stable in a wide range of salt concentrations unlike most of the halophilic lipases 31, 32 . The lipase preserved its activity even at low salt concentrations.…”

Characterization and Screening of Lipolytic Bacteria from Thai Fermented Fish

“…Many microorganisms are found in fermented fish products that can produce extracellular lipases that catalyze the hydrolysis of triacylglycerols to glycerol and free fatty acids, including low molecular weight volatile fatty acids, such as acetic acid, propionic acid and butyric acid (Fukami et al 2002). These later components are associated with the aroma and flavor in fermented fish products (Camacho et al 2009). The aim of this research was to isolate, identify and screen the lipase activity of halophilic bacterial strains from Thai fermented fish products.…”

“…(Gupta et al 2004), Geobacillus sp. TW1 (Li & Zhang 2005), Haloarcula marismortui (Camacho et al 2009), Bacillus sp. strain DVL2 (Kumar et al 2012), B. mesenterichs, B. subtilis, B. cereus (Miettinen et al 2013) and Idiomarina sp.…”