Belo, Isabel

Braga, Adelaide Correia

Bujna, Erika

Farkas, Csilla

Ferreira, Patrícia

Lopes, Marlene

Nagy, Edina Szandra

Nguyen, Quang D.

Sipiczki, Gizella

English

Universidade do Minho: RepositoriUM

XX EuroFoodChem Conference Su_P11  POSTER    284  Enhancement of lipase production by Yarrowia divulgata  Edina Szandra Nagy1*, Erika Bujna1, Gizella Sipiczki1, Csilla Farkas1, Isabel Belo2, Marlene Lopes2, Adelaide Braga2, Patrícia Ferreira2, Quang Duc Nguyen1 1Research Centre for Bioengineering and Process Engineering, Szent István University, Ménesi út 45, H-1118 Budapest, Hungary 2CEB - Centre of Biological Engineering, University of Minho, 4710-057 Braga, Portugal * Nagy.Edina.Szandra@etk.szie.hu   Lipase is a highly valuable compound which has a wide range of usability, for example in the food-, pharmaceutical- and beauty industries. Yarrowia lipolytica is known from its remarkably high lipolytic and proteolytic activity. In the last years some novel species belonging to the Yarrowia genus were described, and some of them also have the ability to produce lipase and other valuable compounds. The main aims of this research were to enhance the lipase production of some novel Yarrowia strains by using olive oil and Tween 80 supplements, and optimizing concentrations of olive oil.  Thirty-five strains of Yarrowia divulgata, Y. porcina and Y. bubula isolated from raw, grounded pork or beef were screened for lipase production by streaking them on the surface of Gorodkowa medium supplemented with olive oil. Inoculum (5%, v/v) was transferred into 500 mL flasks, containing 200 mL of fermentation medium (2% glucose, 0.64% peptone, 1% yeast extract) and supplemented with 1%, 5%, 10%, 20% and 50% olive oil and 0.05% Tween 80. Experiments were carried out for 72 or 148 hours at 28°C in orbital shaker (160 rpm). Samples were centrifuged and supernatants were used for measurement of extracellular lipase activity; additionally, yeast cells were disrupted before the quantification of intracellular lipase. 25mM p-nitrophenyl-laurate was used as substrate and the reaction was performed at 37°C in phosphate buffer (pH 7.2) for 10 min. Lipase activity was assayed spectrophotometrically at 405 nm. One unit (U) of lipase activity was defined as the amount of enzyme that releases 1 μM of p-nitrophenol per minute (pH 7.2, 37°C).  Almost all examined Yarrowia strain showed lipolytic activity, but Y. divulgata 5257 and 2062 were selected for further experiments. In the cases of Y. divulgata strains, 4.03 U/l and 8.11 U/l of extracellular enzyme activities were attained after 8 h and 48 h, respectively. Olive oil and Tween 80 have been published (Corzo & Revah, 1999; Galvagno et al., 2011) to enhance lipase activity and in our experiments this positive effect has also been proved: maximal lipase activity rised to 6,59 U/l and 26.00 U/l at 48 and 64 hours in the prescence of 1% olive oil, and to 83.47 U/l and 203.61 U/l at 72 hours when both additives have been added. The effect of olive oil concentration was also dinvestigated. Y. divulgata 2062 showed the highest extra- and intracellular lipase activity in the prescence of 10% olive oil, although in case of Y. divulgata 5257, 5 and 50% olive oil supplementation was the most successful in case of both intra- and extracellular activity, which is controversial to previously published data (Pignède et al., 2000; Coca & Dustet, 2006). Y. divulgata 5257 showed hight intracellular lipase activity (924.44 U/l), and Y. divulgata 156.73 U/l without Tween 80.  Y. divulgata strains exhibited the best performance and the addition of proper concentration of olive oil and Tween 80 led to the increase of lipase production. Thus besides Y. lipolytica members of other species may have great industrial potential and are worth to study.    Acknowledgments: This work was supported by National Research, Development and Innovation Office through project No. TÉT_16-1-2016-0084 as well as project No. EFOP-3.6.3-VEKOP-16-2017-00005 and bilateral cooperation project FCT/NKFIH.  References: [1] G. Corzo, S. Revah: Bioresource Technology, 70(2) (1999) 173-180. [2] M. A. Galvagno, L. J. Iannone, J. Bianchi, F. Kronberg, E. Rost, M. R. Carstens, P. Cerrutti. Rev Argent Microbiol., 43(3) (2011) 218-25. [3] G. Pignède,H. Wang, F. Fudalej, C. Gaillardin, M. Seman, J. M. Nicaud. Journal of Bacteriology, 182(10) (2000) 2802-2810. [4] J. Coca, J. C. Dustet. Biotecnología Aplicada, 23 (2006) 224-228.  

Enhancement of lipase production by Yarrowia divulgata

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Enhancement of lipase production by Yarrowia divulgata

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