Abstract
The yeast Yarrowia lipolytica possesses multiple paralogues of genes dedicated to hydrophobic substrates metabolization. Among them, 16 lipase encoding genes, involved in lipid or grease breakdown, were highlighted in the yeast genome. However, little information on all those paralogues has been yet obtained. Microarray data suggest that only a few of them could be expressed. Lipase synthesis seems to be dependent on the fatty acid or oil used as carbon source confirming the high adaptation of Y. lipolytica to different hydrophobic substrate. This review focuses on the biochemical characterization of those enzymes with special emphasis on the Lip2p lipase which is the isoenzyme mainly synthesized by Y. lipolytica. The 3D structure of this lipase established by homology modeling confirms that Lip2p is a lipase sensu stricto with a lid covering the active site of the enzyme in its closed conformation. Recent findings on enzyme conditioning in dehydrated or liquid formulation and in enzyme immobilization by entrapment in natural polymers from either organic or mineral origins are also discussed together with long-term storage strategies.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Alloue W, Destain J, Amighi K, Thonart P (2007) Storage of Yarrowia lipolytica lipase after spray-drying in the presence of additives. Process Biochem 42:1357–1361
Alloue W, Destain J, Ongena M, Blecker C, Thonart P (2008) Effect of monopropylene glycol and gamma irradiation on Yarrowia lipolytica lipase stabilization. Prep Biochem Biotechnol 38:217–228
Aloulou A, Rodriguez JA, Puccinelli D, Mouz N, Leclaire J, Leblond Y, Carriere F (2007) Purification and biochemical characterization of the LIP2 lipase from Yarrowia lipolytica. Biochim Biophys Acta 1771:228–237
Bigey F, Tuery K, Bougard D, Nicaud JM, Moulin G (2003) Identification of a triacylglycerol lipase gene family in Candida deformans: molecular cloning and functional expression. Yeast 20:233–248
Bordes F, Cambon E, Dossat-Letisse V, Andre I, Croux C, Nicaud JM, Marty A (2009) Improvement of Yarrowia lipolytica lipase enantioselectivity by using mutagenesis targeted to the substrate binding site. Chembiochem 10:1705–1713
Brockerhoff H (1971) On the function of bile salts and proteins as cofactors of lipase. J Biol Chem 246:5828–5831
Carriere F, Thirstrup K, Hjorth S, Ferrato F, Nielsen PF, Withers-Martinez C, Cambillau C, Boel E, Thim L, Verger R (1997) Pancreatic lipase structure-function relationships by domain exchange. Biochemistry 36:239–248
Casaregola S, Neuveglise C, Lepingle A, Bon E, Feynerol C, Artiguenave F, Wincker P, Gaillardin C (2000) Genomic exploration of the hemiascomycetous yeasts: 17. Yarrowia lipolytica. FEBS Lett 487:95–100
Corzo G, Revah S (1999) Production and characteristics of lipase from Yarrowia lipolytica 681. Bioresour Technol 70:173–180
Desfougeres T, Haddouche R, Fudalej F, Neuveglise C, Nicaud JM (2010) SOA genes encode proteins controlling lipase expression in response to triacylglycerol utilization in the yeast Yarrowia lipolytica. FEMS Yeast Res 10:93–103
Destain J (1998) Production, purification et caractérisation de la lipase de Yarrowia lipolytica. PhD thesis, Faculté Universitaire des Sciences Agronomiques de Gembloux, Gembloux, Belgium
Destain J, Roblain D, Thonart P (1997) Improvement of lipase production from Yarrowia lipolytica. Biotechnol Lett 19:105–107
Destain J, Fickers P, Weekers F, Moreau B, Thonart P (2005) Utilization of methyl oleate in production of microbial lipase. Appl Biochem Biotechnol 121–124:269–277
Dokic P, Jakovljevic J, Dokic-Baucal L (1998) Molecular characteristics of maltodextrin and rheological behaviour of diluted and concentrated solutions. Colloids Surf A 141:435–440
Dominguez A, Costas M, Longo MA, Sanroman A (2003) A novel application of solid state culture: production of lipases by Yarrowia lipolytica. Biotechnol Lett 25:1225–1229
Dujon B, Sherman D, Fischer G, Durrens P, Casaregola S, Lafontaine I, De Montigny J, Marck C, Neuveglise C, Talla E, Goffard N, Frangeul L, Aigle M, Anthouard V, Babour A, Barbe V, Barnay S, Blanchin S, Beckerich JM, Beyne E, Bleykasten C, Boisrame A, Boyer J, Cattolico L, Confanioleri F, De Daruvar A, Despons L, Fabre E, Fairhead C, Ferry-Dumazet H, Groppi A, Hantraye F, Hennequin C, Jauniaux N, Joyet P, Kachouri R, Kerrest A, Koszul R, Lemaire M, Lesur I, Ma L, Muller H, Nicaud JM, Nikolski M, Oztas S, Ozier-Kalogeropoulos O, Pellenz S, Potier S, Richard GF, Straub ML, Suleau A, Swennen D, Tekaia F, Wesolowski-Louvel M, Westhof E, Wirth B, Zeniou-Meyer M, Zivanovic I, Bolotin-Fukuhara M, Thierry A, Bouchier C, Caudron B, Scarpelli C, Gaillardin C, Weissenbach J, Wincker P, Souciet JL (2004) Genome evolution in yeasts. Nature 430:35–44
Fickers P, Nicaud JM (2013) Biotechnological applications of Yarrowia lipolytica lipases: an overview. In: Barth G (ed) Yarrowia lipolytica. Springer, Heidelberg
Fickers P, Nicaud JM, Destain J, Thonart P (2003) Overproduction of lipase by Yarrowia lipolytica mutants. Appl Microbiol Biotechnol 63:136–142
Fickers P, Nicaud JM, Gaillardin C, Destain J, Thonart P (2004) Carbon and nitrogen sources modulate lipase production in the yeast Yarrowia lipolytica. J Appl Microbiol 96:742–749
Fickers P, Benetti PH, Wache Y, Marty A, Mauersberger S, Smit MS, Nicaud JM (2005a) Hydrophobic substrate utilisation by the yeast Yarrowia lipolytica, and its potential applications. FEMS Yeast Res 5:527–543
Fickers P, Fudalej F, Le Dall MT, Casaregola S, Gaillardin C, Thonart P, Nicaud JM (2005b) Identification and characterisation of LIP7 and LIP8 genes encoding two extracellular triacylglycerol lipases in the yeast Yarrowia lipolytica. Fungal Genet Biol 42:264–274
Fickers P, Fudalej F, Nicaud JM, Destain J, Thonart P (2005c) Selection of new over-producing derivatives for the improvement of extracellular lipase production by the non-conventional yeast Yarrowia lipolytica. J Biotechnol 115:379–386
Fickers P, Nicaud JM, Destain J, Thonart P (2005d) Involvement of hexokinase Hxk1 in glucose catabolite repression of LIP2 encoding extracellular lipase in the yeast Yarrowia lipolytica. Curr Microbiol 50:133–137
Fickers P, Ongena M, Destain J, Weekers F, Thonart P (2006) Production and down-stream processing of an extracellular lipase from the yeast Yarrowia lipolytica. Enzyme Microb Technol 38:756–759
Fickers P, Destain J, Thonart P (2009) Improvement of Yarrowia lipolytica lipase production by fed-batch fermentation. J Basic Microbiol 49:212–215
Gombert A, Pinto A, Castilho L, Freire D (1999) Lipase production by Penicillium restrictum in solid state fermentation. Process Biochem 35:85–90
Guerzoni ME, Lanciotti R, Vannini L, Galgano F, Favati F, Gardini F, Suzzi G (2001) Variability of the lipolytic activity in Yarrowia lipolytica and its dependence on environmental conditions. Int J Food Microbiol 69:79–89
Han D, Rhee JS (1986) Characteristics of lipase-catalyzed hydrolysis of olive oil in AOT-isooctane reversed micelles. Biotechnol Bioeng 28:1250–1255
Han K, Levenspiel O (1988) Extended monod kinetics for substrate, product, and cell inhibition. Biotechnol Bioeng 32:430–447
Jolivet P, Bordes F, Fudalej F, Cancino M, Vignaud C, Dossat V, Burghoffer C, Marty A, Chardot T, Nicaud JM (2007) Analysis of Yarrowia lipolytica extracellular lipase Lip2p glycosylation. FEMS Yeast Res 7:1317–1327
Lupescu I, Groposila-Constantinescu D, Jurcoane S, Diguta C, Cozea A, Tcacenco L (2007) Production of lipase by a strain of the non-conventional yeast Yarrowia lipolytica and isolation of crude enzyme. Rom Biotechnol Lett 12:27–37
Mauersberger S, Wang HJ, Gaillardin C, Barth G, Nicaud JM (2001) Insertional mutagenesis in the n-alkane-assimilating yeast Yarrowia lipolytica: generation of tagged mutations in genes involved in hydrophobic substrate utilization. J Bacteriol 183:5102–5109
Mekouar M, Blanc-Lenfle I, Ozanne C, Da Silva C, Cruaud C, Wincker P (2010) Detection and analysis of alternative splicing in Yarrowia lipolytica reveal structural constraints facilitating nonsense-mediated decay of intron-retaining transcripts. Genome Biol 11:R65
Nga B, Gaillardin C, Fournier P, Heslot H (1989) Genetic analysis of lipase low producing mutant of Yarrowia lipolytica. J Gen Microbiol 135:2439–2443
Noel M, Lozano P, Combes D (2005) Polyhedric alcohol protective effect on Rhizomucor miehei lipase deactivation enhanced by pressure and temperature treatment. Bioprocess Biosyst Eng 27:375–380
Novotny C, Dolezalova L, Musil P, Novak M (1988) The production of lipase by some Candida and Yarrowia yeast. J Basic Microbiol 4:221–227
Novotny C, Dolezalova L, Lieblova J (1994) Dimorphic growth and lipase production in lipolytic yeasts. Folia Microbiol (Praha) 39:71–73
Ollis DL, Cheah E, Cygler M, Dijkstra B, Frolow F, Franken SM, Harel M, Remington SJ, Silman I, Schrag J (1992) The alpha/beta hydrolase fold. Protein Eng 5:197–211
Ota Y, Nakamiya T, Yamada K (1970) Lipase from Candida paralipolytica: purification, some properties and modification of the purified enzyme with concentrated solution of sodium chloride. Agric Biol Chem 34:1368–1374
Ota Y, Gomi S, Morimoto Y, Minod Y (1982) Purification and some properties of cell-bound lipase from Saccharomycopsis lipolytica. Agric Biol Chem 46:2885–2893
Peireira-Meirelles F, Rocha L, Sant’Anna G (1997) A stable lipase from Candida lipolytica : cultivation conditions and crude enzyme characteristics. Appl Biochem Biotechnol 63–65:73–85
Pignede G, Wang H, Fudalej F, Gaillardin C, Seman M, Nicaud JM (2000a) Characterization of an extracellular lipase encoded by LIP2 in Yarrowia lipolytica. J Bacteriol 182:2802–2810
Pignede G, Wang HJ, Fudalej F, Seman M, Gaillardin C, Nicaud JM (2000b) Autocloning and amplification of LIP2 in Yarrowia lipolytica. Appl Environ Microbiol 66:3283–3289
Pleiss J, Scheib H, Schmid RD (2000) The His gap motif in microbial lipases: a determinant of stereoselectivity toward triacylglycerols and analogs. Biochimie 82:1043–1052
Sarda L, Desnuelle P (1958) Actions of pancreatic lipase on esters in emulsions. Biochim Biophys Acta 30:513–521
Song H, Jiang Z, Ma L (2006) Expression and purification of two lipase from Yarrowia lipolytica AS 2.1216. Protein Expr Purif 47:393–397
Suzuki T, Imamura K, Yamamoto K, Satoh T, Okazaki M (1997) Thermal stabilization of freeze-dried enzymes by sugars. J Chem Eng Jpn 30:609–613
Tanaka A, Sugimoto H, Muta Y, Mizuno T, Senoo K, Obata H (2003) Differential scanning calorimetry of the effects of calcium on the thermal unfolding of Pseudomonas cepacia lipase. Biosci Biotechnol Biochem 67:207–210
Theveniau F, Gaillardin C, Nicaud JM (2009) Application of the non-conventional yeast Yarrowia lipolytica. In: Satyanarayana T, Kunze G (eds) Yeast biotechnology: diversity and applications. Springer, Dordrecht, pp 589–613
Thevenieau F, Le Dall MT, Nthangeni B, Mauersberger S, Marchal R, Nicaud JM (2007) Characterization of Yarrowia lipolytica mutants affected in hydrophobic substrate utilization. Fungal Genet Biol 44:531–542
Turki S, Kraeim IB, Weeckers F, Thonart P, Kallel H (2009) Isolation of bioactive peptides from tryptone that modulate lipase production in Yarrowia lipolytica. Bioresour Technol 100:2724–2731
Turki S, Ayed A, Chalghoumi N, Weekers F, Thonart P, Kallel H (2010) An enhanced process for the production of a highly purified extracellular lipase in the non-conventional yeast Yarrowia lipolytica. Appl Biochem Biotechnol 160:1371–1385
Verger R (1997) Interfacial activation of lipase: facts and artifacts. Trends Biotechnol 15:32–38
Yu M, Lange S, Richter S, Tan T, Schmid RD (2007a) High-level expression of extracellular lipase Lip2 from Yarrowia lipolytica in Pichia pastoris and its purification and characterization. Protein Expr Purif 53:255–263
Yu M, Qin S, Tan T (2007b) Purification and characterisation of the extracellular lipase Lip2 from Yarrowia lipolytica. Process Biochem 42:384–391
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer-Verlag Berlin Heidelberg
About this chapter
Cite this chapter
Fickers, P., Sauveplane, V., Nicaud, JM. (2013). The Lipases from Y. lipolytica: Genetics, Production, Regulation, and Biochemical Characterization. In: Barth, G. (eds) Yarrowia lipolytica. Microbiology Monographs, vol 25. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-38583-4_3
Download citation
DOI: https://doi.org/10.1007/978-3-642-38583-4_3
Published:
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-38582-7
Online ISBN: 978-3-642-38583-4
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)