Grasas y Aceites, Vol 70, No 4 (2019)

Monitoring of the enzymatic activity of intracellular lipases of Ustilago maydis expressed during the growth under nitrogen limitation and its correlation in lipolytic reactions

M. G. Araiza-Villanueva
Instituto Politécnico Nacional. Escuela Nacional de Ciencias Biológicas, Departamento de Microbiología, Mexico

D. R. Olicón-Hernández
Instituto Politécnico Nacional. Escuela Nacional de Ciencias Biológicas, Departamento de Microbiología - Universidad Nacional Autónoma de México, Facultad de Medicina, Departamento de Bioquímica, Mexico

J. P. Pardo
Universidad Nacional Autónoma de México, Facultad de Medicina, Departamento de Bioquímica, Mexico

H. Vázquez-Meza
Universidad Nacional Autónoma de México, Facultad de Medicina, Departamento de Bioquímica, Mexico

G. Guerra-Sánchez
Instituto Politécnico Nacional. Escuela Nacional de Ciencias Biológicas, Departamento de Microbiología, Mexico


Under nitrogen starvation, Ustilago maydis forms lipid droplets (LDs). Although the dynamics of these organelles are known in the literature, the identity of the lipases implicated in their degradation is unknown. We determined lipase activity and identified the intracellular lipases expressed during growth under nitrogen starvation and YPD media by zymograms. The results showed that cytosolic extracts exhibited higher lipase activity when cells were grown in YPD. Under nitrogen starvation, lipase activity was not detected after 24 h of culture, resulting in lipid accumulation in LDs. This suggests that these lipases could be implicated in LD degradation. In the zymogram, two bands, one of 25 and the other of 37 kDa, presented lipase activity. The YPD extracts showed lipase activity in olive and almond oils, which contain triacylglycerols with mono and polyunsaturated fatty acids. This is the first report about U. maydis cytosolic lipases involved in LD degradation.


Cytosolic lipases; Fatty acids; Lipid droplets; Lipid droplet index; Nitrogen starvation; Triacylglycerol

Full Text:



Athenstaedt K, Daum G. 2003. YMR313c/TGL3 encodes a novel triacylglycerol lipase located in lipid particles of Saccharomyces cerevisiae. J. Biol. Chem. 278, 23317–23323.

Athenstaedt K, Daum G. 2005. Tgl4p and Tgl5p, two triacylglycerol lipases of the yeast Saccharomyces cerevisiae are localized to lipid particles. J. Biol. Chem. 280, 37301–37309.

Benarouche A, Point V, Carriere F, Cavalier JF. 2014. An interfacial and comparative in vitro study of gastrointestinal lipases and Yarrowia lipolytica LIP2 lipase, a candidate for enzyme replacement therapy. Biochimie 102, 145–153.

Berhanu A, Amare G. 2012. Microbial lipases and their industrial applications: Review. Biotechnology 11, 100–118.

Bermudez B, Lopez S, Ortega A, Varela LM, Pacheco YM, Abia R, Muriana FJG. 2011. Oleic acid in olive oil: from a metabolic framework toward a clinical perspective. Current Pharmaceutical Design 17, 831–843.

Bozaquel-Morais BL, Madeira JB, Maya-Monteiro CM, Masuda CA, Montero-Lomeli M. 2010. A new fluorescence-based method identifies protein phosphatases regulating lipid droplet metabolism. PLoS One 5, e13692.

Brabcova J, Prchalova D, Demianova Z, Bucankova A, Vogel H, Valterova I, Pichova I, Zarevucka M. 2013. Characterization of neutral lipase BT-1 isolated from the labial gland of Bombus terrestris males. PLoS One 8, e80066.

Brundiek H, Sass S, Evitt A, Kourist R, Bornscheuer UT. 2012. The short form of the recombinant CAL-A-type lipase UM03410 from the smut fungus Ustilago maydis exhibits an inherent trans-fatty acid selectivity. Appl. Microbiol. Biotechnol. 94, 141–150.

Buerth C, Kovacic F, Stock J, Terfruchte M, Wilhelm S, Jaeger KE, Feldbrugge M, Schipper K, Ernst JF, Tielker D. 2014. Uml2 is a novel CalB-type lipase of Ustilago maydis with phospholipase A activity. Appl. Microbiol. Biotechnol. 98, 4963–4973.

Chavan S, Smith SM. 2014. A rapid and efficient method for assessing pathogenicity of Ustilago maydis on maize and teosinte lines. J. Vis. Exp. 83. e50712.

Fickers P, Marty A, Nicaud JM. 2011. The lipases from Yarrowia lipolytica: genetics, production, regulation, biochemical characterization and biotechnological applications. Biotechnol. Adv. 29, 632–644.

Givianrad MH, Saber-Tehrani M, Jafari Mohammadi SA. 2013. Chemical composition of oils from wild almond (Prunus scoparia) and wild pistachio (Pistacia atlantica). Grasas Aceites 64 (1), 77–84.

Grillitsch K, Connerth M, Kofeler H, Arrey TN, Rietschel B, Wagner B, Karas M, Daum G. 2011. Lipid particles/ droplets of the yeast Saccharomyces cerevisiae revisited: lipidome meets proteome. Biochim. Biophys. Acta 1811, 1165–1176.

Gupta N, Rathi P, Gupta R. 2002. Simplified para-nitrophenyl palmitate assay for lipases and esterases. Analytical Biochemistry 311, 98–99.

Kanwar SS, Kaushal RK, Jawed A, Gupta R, Chimni SS. 2005. Methods for inhibition of residual lipase activity in colorimetric assay: a comparative study. Indian J. Biochem. Biophys. 42, 233–237.

Kerkhoven EJ, Pomraning KR, Baker SE, Nielsen J. 2016. Regulation of amino-acid metabolism controls flux to lipid accumulation in Yarrowia lipolytica. Npj Systems Biology and Applications 2, 16005.

Klug L, Daum G. 2014. Yeast lipid metabolism at a glance. FEMS Yeast Res. 14, 369–388.

Kouker G, Jaeger KE. 1987. Specific and sensitive plate assay for bacterial lipases. Appl. Environ. Microbiol. 53, 211–213.

Kumar D, Kumar L, Nagar S, Raina C, Parshad R, Gupta V. 2012. Screening, isolation and production of lipase/esterase producing Bacillus sp. strain DVL2 and its potential evaluation in esterification and resolution reactions Arch. Appl. Sci. Res. 4, 1763–1770.

Li D, Song JZ, Li H, Shan MH, Liang Y, Zhu J, Xie Z. 2015. Storage lipid synthesis is necessary for autophagy induced by nitrogen starvation. FEBS Lett. 589, 269–276.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275.

Manda NK, Thunuguntla VBSC, Bokka C, Singh BJ. 2017. Ymr210wp leads to the accumulation of phospholipids and steryl esters in yeast. Bioinformation 13, 360–365.

Ortega A, Varela LM, Bermudez B, Lopez S, Muriana FJG, Abia R. 2012. Nutrigenomics and atherosclerosis: The postprandial and long-term effects of virgin olive oil ingestion, in Parthasarathy S (ed) Atherogenesis. IntechOpen, Shangai, 135–160.

Paulino BN, Pessoa MG, Molina G, Kaupert Neto AA, Oliveira JVC, Mano MCR, Pastore GM. 2017. Biotechnological production of value-added compounds by ustilaginomycetous yeasts. Appl. Microbiol. Biotechnol. 101, 7789–7809.

Perez D, Martin S, Fernandez-Lorente G, Filice M, Guisan JM, Ventosa A, Garcia MT, Mellado E. 2011. A novel halophilic lipase, LipBL, showing high efficiency in the production of eicosapentaenoic acid (EPA). PLoS One 6, e23325.

Rambold AS, Cohen S, Lippincott-Schwartz J. 2015. Fatty acid trafficking in starved cells: regulation by lipid droplet lipolysis, autophagy, and mitochondrial fusion dynamics. Dev. Cell 32, 678–692.

Romero-Aguilar L, Pardo JP, Montero-Lomeli M, Luqueño- Bocardo OI, Juarez Oropeza MA, Guerra Sanchez G. 2017. Lipid droplets accumulation and other biochemical changes induced in the fungal pathogen Ustilago maydis under nitrogen-starvation. Arch. Microbiol. 199, 1195–1209.

Roncero JM, Álvarez-Ortí M, Pardo-Giménez A, Gómez R, Rabadán A, Pardo JE. 2016. Virgin almond oil: Extraction methods and composition. Grasas Aceites 67, e143.

Saavedra E, Ramos-Casillas LE, Marin-Hernandez A, Moreno- Sanchez R, Guerra-Sanchez G. 2008. Glycolysis in Ustilago maydis. FEMS Yeast Res. 8, 1313–1323.

Schmidt C, Athenstaedt K, Koch B, Ploier B, Korber M, Zellnig G, Daum G. 2014. Defects in triacylglycerol lipolysis affect synthesis of triacylglycerols and steryl esters in the yeast. Biochim. Biophys. Acta 1842, 1393–1402.

Snellman EA, Sullivan ER, Colwell RR. 2002. Purification and properties of the extracellular lipase, LipA, of Acinetobacter sp. RAG-1. European Journal of Biochemistry 269, 5771–5779.

Ugur A, Sarac N, Boran R, Ayaz B, Ceylan O, Okmen G. 2014. New lipase for biodiesel production: Partial purification and pharacterization of LipSB 25–4. ISRN Biochem. 2014, 289749.

Vingering N, Oseredczuk M, du Chaffaut L, Ireland J, Ledoux M. 2010. Fatty acid composition of commercial vegetable oils from the French market analysed using a long highly polar column. OCL 17, 185–192.

Welte MA. 2015. Expanding roles for lipid droplets. Curr. Biol. 25, R470–481.

Yanty NAM, Marikkar JMN, Long K. 2011. Effect of varietal differences on composition and thermal characteristics of avocado oil. J.A.O.C.S. 88, 1997–2003.

Zavala-Moreno A, Arreguin-Espinosa R, Pardo JP, Romero- Aguilar L, Guerra-Sánchez G. 2014. Nitrogen source affects glycolipid production and lipid accumulation in the phytopathogen fungus Ustilago maydis. Advances in Microbiology 4, 934–944.

Zhu Z, Ding Y, Gong Z, Yang L, Zhang S, Zhang C, Lin X, Shen H, Zou H, Xie Z, Yang F, Zhao X, Liu P, Zhao ZK. 2015. Dynamics of the lipid droplet proteome of the oleaginous yeast Rhodosporidium toruloides. Eukaryot Cell 14, 252–264.

Copyright (c) 2019 Consejo Superior de Investigaciones Científicas (CSIC)

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.

Contact us

Technical support