The aim of this study was to characterize lipases from two thermophilic bacteria, <i> Geobacillus stearothermophilus </i> (GS) and <i> Anoxybacillus flavithermus </i> (AF) in heat treated cell lysates. The pH optimum, pH stability, temperature stability and substrate kinetics and specificity of the lipases w ere determined. Optimum activity of the lipase from GS (LGS) was observed at pH 7.5, and the optimum activity of the lipase from AF (LAF) was at pH 8.0. LGS was stable up to 70 ° C after 12 hrs w hile LAF was stable up to 90 ° C after 12 hrs. Both enzymes were stable at a pH range of 6 to 8 over 12 h at 4 ° C . LGS had a highest <i> V max </i> value of 22 mM · min - 1 · mg - 1 with p-nitrophenyl acetate while the lowest <i> K m </i> was 0.8 mM with p-nitrophenyl laurate. The highest <i> V max </i> of LAF was 2.5 mM · min - 1 · mg - 1 with p-nitrophenyl myristate, and the lowest <i> K m </i> was 0.4 mM with p-nitrophenyl octanoate. LGS preferentially hydrolyzed p-nitrophenyl acetate and p-nitrophenyl octanoate while LAF preferentially hydrolyzed p-nitrophenyl myristate and p-nitrophenyldodecanoate. Lipases from both GS and AF showed characteristics that would be beneficial in food processing.
Chandra, P., Enespa, S.R. and Arora, P.K. (2020) Microbial Lipases and Their Industrial Applications: A Comprehensive Review. Microbial Cell Factories, 19, 169. https://doi.org/10.1186/s12934-020-01428-8
Ekinci, A.P., Dincer, B., Baltas, N. and Adigüzel, A. (2016) Partial Purification and Characterization of Lipase from Geobacillus stearothermophilus AH22. Journal of Enzyme Inhibition and Medicinal Chemistry, 31, 325-331. https://doi.org/10.3109/14756366.2015.1024677
Nazina, T., Tourova, T., Poltaraus, A., Novikova, E., Grigoryan, A., Ivanova, A., Lysenko, A., Petrunyaka, V., Osipov, G. and Belyaev, S. (2001) Taxonomic Study of Aerobic Thermophilic Bacilli: Descriptions of Geobacillus subterraneus gen. nov., sp. nov. and Geobacillusuzenensis sp. nov. from Petroleum Reservoirs and Transfer of Bacillus stearothermophilus, Bacillus thermocatenulatus, Bacillus thermoleovorans, Bacillus kaustophilus, Bacillus thermodenitrificans to Geobacillus as the New Combinations G. stearothermophilus, G. thermocatenulatus, G. thermoleovorans, G. kaustophilus, G. thermoglucosidasius and G. thermodenitrificans. International Journal of Systematic and Evolutionary Microbiology, 51, 433-446. https://doi.org/10.1099/00207713-51-2-433
Pikuta, E., Lysenko, A., Chuvilskaya, N., Mendrock, U., Hippe, H., Suzina, N., Nikitin, D., Osipov, G. and Laurinavichius, K. (2000) Anoxybacillus pushchinensis gen. nov., sp. nov., a Novel Anaerobic, Alkaliphilic, Moderately Thermophilic Bacterium from Manure, and Description of Anoxybacillus flavithermus comb. nov. International Journal of Systematic and Evolutionary Microbiology, 50, 2109-2117. https://doi.org/10.1099/00207713-50-6-2109
Coelho, A.L.S. and Orlandelli, R.C. (2020) Immobilized Microbial Lipases in the Food Industry: A Systematic Review. Critical Reviews in Food Science and Nutrition, 61, 1689-1703. https://doi.org/10.1080/10408398.2020.1764489
Guerrand, D. (2017) Lipases Industrial Applications: Focus on Food and Agroindustries. OCL, 24, D403. https://doi.org/10.1051/ocl/2017031
DiCosimo, R., McAuliffe, J., Poulose, A.J. and Bohlmann, G. (2013) Industrial Use of Immobilized Enzymes. Chemical Society Reviews, 42, 6437-6474. https://doi.org/10.1039/c3cs35506c
Ewis, H.E., Abdelal, A.T. and Lu, C.D. (2004) Molecular Cloning and Characterization of Two Thermostable Carboxyl Esterases from Geobacillus stearothermophilus. Gene, 329, 187-195. https://doi.org/10.1016/j.gene.2003.12.029
Bakir, Z.B. and Metin, K. (2016) Purification and Characterization of an Alkali-Thermostable Lipase from Thermophilic Anoxybacillus flavithermus HBB 134. Journal of Microbiology and Biotechnology, 26, 1087-1097. https://doi.org/10.4014/jmb.1512.12056
Beatty, N.F. and Walsh, M.K. (2016) Influence of Thermosonication on Geobacillus stearothermophilus Inactivation in Skim Milk. International Dairy Journal, 61, 10-17. https://doi.org/10.1016/j.idairyj.2016.03.011
Saw, J.H., Mountain, B.W., Feng, L., Omelchenko, M.V., Hou, S., Saito, J.A., Stott, M.B., Zhao, D., Li, G. and Wu, J. (2008) Encapsulated in Silica: Genome, Proteome, and Physiology of the Thermophilic Bacterium Anoxybacillus flavithermus WK1. Genome Biology, 9, R161. https://doi.org/10.1186/gb-2008-9-11-r161
Lee, D.W., Kim, H.W., Lee, K.W., Kim, B.C., Choe, E.A., Lee, H.S., Kim, D.S. and Pyun, Y.R. (2001) Purification and Characterization of Two Distinct Thermostable Lipases from the Gram-Positive Thermophilic Bacterium Bacillus thermoleovorans ID-1. Enzyme and Microbial Technology, 29, 363-371. https://doi.org/10.1016/S0141-0229(01)00408-2
Glogauer, A., Martini, V.P., Faoro, H., Couto, G.H., Müller-Santos, M., Monteiro, R.A., Mitchell, D.A., de Souza, E.M., Pedrosa, F.O. and Krieger, N. (2011) Identification and Characterization of a New True Lipase Isolated through Metagenomic Approach. Microbial Cell Factories, 10, 54. https://doi.org/10.1186/1475-2859-10-54
Najm, T.A. and Walsh, M.K. (2021) Comparison of Four Purification Methods to Purify Lipases from Thermophilic Bacteria. BABT, 64, e21200045.
Jiang, Y., Zhou, X. and Chen, Z. (2010) Cloning, Expression, and Biochemical Characterization of a Thermostable Lipase from Geobacillus stearothermophilus JC. World Journal of Microbiology and Biotechnology, 26, 747-751. https://doi.org/10.1007/s11274-009-0213-1
Ay, F., Karaoglu, H., Inan, K., Canakci, S. and Belduz, A.O. (2011) Cloning, Purification, and Characterization of a Thermostable Carboxylesterase from Anoxybacillus sp. PDF1. Protein Expression and Purification, 80, 74-79. https://doi.org/10.1016/j.pep.2011.06.019
Colak, A., Sisik, D., Saglam, N., Güner, S., Canakci, S. and Beldüz, A.O. (2005) Characterization of a Thermoalkalophilic Esterase from a Novel Thermophilic Bacterium, Anoxybacillus gonensis G2. Bioresource Technology, 96, 625-631. https://doi.org/10.1016/j.biortech.2004.06.003
Chen, P.T., Lin, C.H., Chen, Y.T., Hsu, F.Y. and Shsw, J.F. (2020) Isolation, Expression, and Characterization of the Thermophilic Recombinant Esterase from Geobacillus thermodenitrificans PS01. Applied Biochemistry and Biotechnology, 191, 112-124. https://doi.org/10.1007/s12010-020-03225-w
Faiz, O., Colak, A., Saglam, N., Canakci, S. and Belduz, A.O. (2007) Determination and Characterization of Thermostableesterolytic Activity from a Novel Thermophilic Bacterium Anoxybacillus gonensis A4. BMB Reports, 40, 588-594. https://doi.org/10.5483/BMBRep.2007.40.4.588
Lee, K.T. and Foglia, T. (2000) Fractionation of Chicken Fat Triacylglycerols: Synthesis of Structured Lipids with Immobilized Lipases. Journal of Food Science, 65, 826-831. https://doi.org/10.1111/j.1365-2621.2000.tb13595.x
Gibon, V. and Kellens, M. (2014) Latest Developments in Chemical and Enzymatic Interesterification for Commodity Oils and Specialty Fats. In: Trans Fats Replacement Solutions, Elsevier, Amsterdam, 153-185. https://doi.org/10.1016/B978-0-9830791-5-6.50013-7
Sharma, A., Sharma, T., Meena, K. and Kanwar, S. (2017) Physical Adsorption of Lipase onto Mesoporous Silica. International Journal of Current Advanced Research, 6, 3836-3841. https://doi.org/10.24327/ijcar.2017.3841.0378
Ebrahimpour, A., Rahman, R., Basri, M. and Salleh, A.B. (2011) High Level Expression and Characterization of a Novel Thermostable, Organic Solvent Tolerant, 1,3-Regioselective Lipase from Geobacillus sp. Strain ARM. Bioresource Technology, 102, 6972-6981. https://doi.org/10.1016/j.biortech.2011.03.083
Tayyab, M., Rashid, N. and Akhtar, M. (2011) Isolation and Identification of Lipase Producing Thermophilic Geobacillus sp. SBS-4S: Cloning and Characterization of the Lipase. Journal of Bioscience and Bioengineering, 111, 272-278. https://doi.org/10.1016/j.jbiosc.2010.11.015
Fucinos, P., González, R., Atanes, E., Sestelo, A.B.F., Pérez-Guerra, N. and Pastrana Rúa, M.L. (2012) Lipases and Esterases from Extremophiles: Overview and Case Example of the Production and Purification of an Esterase from Thermus thermophilus HB27. Methods in Molecular Biology, 861, 239-266. https://doi.org/10.1007/978-1-61779-600-5_15
Romero, C.M., Pera, L.M., Loto, F., Vallejos, C., Castro, G. and Baigori, M.D. (2012) Purification of an Organic Solvent-Tolerant Lipase from Aspergillus niger MYA 135 and Its Application in Ester Synthesis. Biocatalysis and Agricultural Biotechnology, 1, 25-31. https://doi.org/10.1016/j.bcab.2011.08.013
Esteban-Torres, M., Barcenilla, J.M., Mancheno, J.M., de las Rivas, B. and Munoz, R. (2014) Characterization of a Versatile Arylesterase from Lactobacillus plantarum Active on Wine Esters. Journal of Agricultural and Food Chemistry, 62, 5118-5125. https://doi.org/10.1021/jf500991m