ENZYMATIC CHARACTERIZATION OF YEAST ISOLATED FROM NATURALLY FERMENTED HERBS

Year 2021, Volume: 46 Issue: 5, 1081 - 1091, 05.08.2021

Erkan Güneş Furkan Aydın İbrahim Çakır

https://doi.org/10.15237/gida.GD21088

Cited By: 2

Abstract

This study aims to identify yeasts from spontaneously fermented herbs used for Van herby cheese and to evaluate their enzymatic activities. Sequencing of partial 26S rRNA gene revealed the species of Pichia membranifaciens (n = 8), Kazachstania servazzii (n = 6), Debaryomyces hansenii (n = 2), Kluyveromyces marxianus (n = 2), and Pichia fermentans (n = 2). Enzyme profiles were determined using API-ZYM strips. The isolates had diverse enzyme activities, including alkaline and acid phosphatase, esterase, esterase lipase, aminopeptidases, and proteases mostly at different levels, which may have crucial roles during ripening of the cheese. K. marxianus M8.1, D. hansenii M6.1, and M6.3, P. membraniciens M13.1, M13.2, and M14.1 had superior and diverse enzymatic characteristics. Such enzymatic activities could be of great technological importance for the determination of adjunct culture along with starter lactic acid bacteria for the production of Van herby cheese.

Keywords

Herby cheese, traditional cheese, yeast, enzyme activity.

Thanks

The authors would like to thank Prof. Dr. Hayri COSKUN and Prof. Dr. Yusuf TUNCTURK for supplying herb brines from Van province of Turkey.

FERMENTE SALAMURA OTLARDAN İZOLE EDİLEN MAYALARIN ENZİMATİK KARAKTERİZASYONU

Abstract

Bu çalışmada, Van otlu peyniri üretiminde kullanılan fermente salamura otlarından mayaların izolasyonu, identifikasyonu ve enzimatik aktivitelerinin belirlenmesi amaçlanmıştır. Yirmi adet maya izolatı izole edilerek moleküler olarak tanımlanmıştır. Tanımlamada 26S rRNA gen bölgesinin D1/D2 alt bölgesi çoğaltılarak sekanslanmıştır. Sekanslama sonucunda Pichia membranifaciens (n = 8), Kazachstania servazzii (n = 6), Debaryomyces hansenii (n = 2), Kluyveromyces marxianus (n = 2) ve Pichia fermentans (n = 2) türleri tanımlanmıştır. İzolatların enzim profilleri API-ZYM test kitleri kullanılarak belirlenmiştir. İzolatların, peynirin olgunlaşması sırasında önemli rollere sahip olabilecek alkali ve asit fosfataz, esteraz, esteraz lipaz, aminopeptidazlar ve proteazlar dahil olmak üzere çeşitli enzim aktivitelerine sahip olduğu belirlenmiştir. İzolatlar arasından, K. marxianus M8.1, D. hansenii M6.1 ile M6.1, P. membraniciens M13.1, M13.2 ile M14.1 suşlarının üstün enzimatik özellikler taşıdığı görülmüştür. Bu tür enzimatik faaliyetlerin, Van otlu peyniri üretimi için starter laktik asit bakterileri ile birlikte ek kültürün belirlenmesi için teknolojik bir öneme sahip olabileceği düşünülmektedir.

Keywords

Otlu peynir, geleneksel peynir, maya, enzim aktivitesi.

References

• Akuzawa, R., P.F. Fox. (2004). Acid phosphatase in cheese. Anim. Sci. 75 (5): 385-391, doi: 10.1111/j.1740-0929.2004.00202.x
• Al-Otaibi, M.M. (2012). Isolation and identification of lactic acid bacteria and yeasts from Sameel milk: a Saudi traditional fermented milk. Int. J. Dairy Sci. 7: 73-83, doi: 10.3923/ijds.2012.73.83
• Álvarez-Martín, P., Flórez, A.B., López-Díaz, T.M., Mayo, B. (2007). Phenotypic and molecular identification of yeast species associated with Spanish blue-veined Cabrales cheese. Int. Dairy J. 17 (8): 961-967, doi: 10.1016/j.idairyj.2006.11.005
• Aponte, M., Pepe, O., Blaiotta, G. (2010). Identification and technological characterization of yeast strains isolated from samples of water buffalo Mozzarella cheese. J. Dairy Sci. 93(6): 2358-2361, doi: 10.3168/jds.2009-2948
• Atanassova, M.R., Fernández-Otero, C., Rodríguez-Alonso, P., Fernández-No, I.C., Garabal, J. I., J., Centeno, A. (2016). Characterization of yeasts isolated from artisanal short-ripened cows' cheeses produced in Galicia (NW Spain). Food Microbiol. 53: 172-181, doi: 10.3168/jds.2009-2948
• Aydin, F., Ozer, G., Alkan, M., Cakir, I. (2020). The utility of iPBS retrotransposons markers to analyze genetic variation in yeast. Int. J. Food Microbiol. 325C: 108647, doi: 10.1016/j.ijfoodmicro.2020.108647
• Banjara, N., Suhr, M.J., Hallen-Adams, H.E. (2015). Diversity of yeast and mold species from a variety of cheese types. Curr. Microbiol. 70(6): 792-800, doi: 10.1007/s00284-015-0790-1
• Biagiotti, C., Ciani, M., Canonico, L., Comitini, F. (2018). Occurrence and involvement of yeast biota in ripening of Italian Fossa cheese. Eur. Food Res. Technol. 244 (11): 1921-1931, doi: 10.1007/s00217-018-3104-6
• Binetti, A., Carrasco, M., Reinheimer, J., Suárez, V. (2013). Yeasts from autochthonal cheese starters: technological and functional properties. J. Appl. Microbiol. 115(2): 434-444, doi: 10.1111/jam.12228
• Gobbetti, M., De Angelis, M., Di Cagno, R., Mancini, L., Fox, P.F. (2015). Pros and cons for using non-starter lactic acid bacteria (NSLAB) as secondary/adjunct starters for cheese ripening. Trends Food Sci. Technol. 45(2): 167-178, doi: 10.1016/j.tifs.2015.07.016
• Cakir I. (2010). Antibacterial and antifungal activities of some lactic acid bacteria isolated from naturally fermented herbs. J Food Agric. Environ. 8: 223-226.
• Cardoso, V.M., Borelli, B.M., Lara, C. A., Soares, M.A., Pataro, C., Bodevan, E.C., Rosa, C.A. (2015). The influence of seasons and ripening time on yeast communities of a traditional Brazilian cheese. Food Res. Int. 69: 331-340, doi: 10.1016/j.foodres.2014.12.040
• Ceugniez, A., Drider, D., Jacques, P., Coucheney, F. (2015). Yeast diversity in a traditional French cheese “Tomme d'orchies” reveals infrequent and frequent species with associated benefits. Food Microbiol. 52: 177-184, doi: 10.1016/j.fm.2015.08.001
• Ceugniez, A., Taminiau, B., Coucheney, F., Jacques, P., Delcenserie, V., Daube, G., Drider. D. (2017). Fungal diversity of “Tomme d'Orchies” cheese during the ripening process as revealed by a metagenomic study. Int. J. Food Microbiol. 258: 89–93, doi: 10.1016/j.ijfoodmicro.2017.07.015
• Coskun, H. (1998). Microbiological and biochemical changes in herby cheese during ripening. Nahrung. 42(5): 309-313, doi: http://dx.doi.org/10.1002/(SICI)1521-3803(199810)42:05%3C309::AID-FOOD309%3E3.0.CO;2-U
• De Freitas, I., Pinon, N., Berdagué, J. L., Tournayre, P., Lortal, S., Thierry, A. (2008). Kluyveromyces lactis but not Pichia fermentans used as adjunct culture modifies the olfactory profiles of Cantalet cheese. J. Dairy Sci. 91(2): 531-543, doi: 10.3168/jds.2007-0136.
• Decimo, M., Brasca, M., Ordóñez, J.A., Cabeza, M.C. (2017). Fatty acids released from cream by psychrotrophs isolated from bovine raw milk. Int. J. Dairy Technol. 70 (3): 339-344, doi: 10.1111/1471-0307.12347
• Dodor, D.E., Tabatabai, M.A. (2007). Arylamidase activity as an index of nitrogen mineralization in soils. Commun. Soil Sci. Plan. 38 (15-16): 2197-2207, doi: 10.1080/00103620701549132
• Dos Santos, M.T.P.G., Benito, M. J., de Guía Córdoba, M., Alvarenga, N. And Herrera, S.R. M.S. (2017). Yeast community in traditional Portuguese Serpa cheese by culture-dependent and-independent DNA approaches. Int. J. Food Microbiol. 262: 63-70, doi: 10.1016/j.ijfoodmicro.2017.09.013
• Dugat-Bony, E., Garnier, L., Denonfoux, J., Ferreira, S., Sarthou, A.S., Bonnarme P., Irlinger, F. 2016. Highlighting the microbial diversity of 12 French cheese varieties. Int. J. Food Microbiol, 238: 265-273, doi: 10.1016/j.ijfoodmicro.2016.09.026
• Frohlich‐Wyder, M.T., Arias‐Roth, E., E. Jakob. (2019). Cheese yeasts. Yeast. 36 (3): 129-141, doi: 10.1002/yea.3368
• Haastrup, M. K., Johansen, P., Malskær, A. H., Castro-Mejía, J. L., Kot, W., Krych, Arneborg, N., Jespersen, L. (2018). Cheese brines from Danish dairies reveal a complex microbiota comprising several halotolerant bacteria and yeasts. Int. J. Food Microbiol. 285: 173-187, doi: 10.1016/j.ijfoodmicro.2018.08.015
• Hayaloglu, A.A., N. Y. Farkye. 2011. Cheese with Added Herbs, Spices and Condiments. In: Encyclopedia of Dairy Sciences, Fuquay JW, Fox PF and McSweeney PLH (eds.), Second Edition, Volume. 1, Academic Press, US, pp. 783–789, doi: 10.1016/B978-0-12-374407-4.00507-0
• Karasu-Yalcin, S., Senses-Ergul, S., Ozbas, Z.Y. (2012). Identification and enzymatic characterization of the yeasts isolated from Erzincan tulum cheese. Mljekarstvo. 62(1): 53-61.
• Karasu-Yalcin, S., Senses-Ergul, S., Ozbas, Z.Y. (2017). Enzymatic characterization of yeast strains originated from traditional Mihalic cheese. J. Microbiol. Biotechnol. Food Sci. 6(5): 1152-1156, doi: 10.15414/jmbfs.2017.6.5.1152-1156
• Kesenkas, H., Akbulut, N. (2008). Yeasts as ripening adjunct cultures in Turkish white brined cheese production. Turk J. Vet. Anim. Sci. 32(5): 327-333.
• Kumar, S., Stecher, G., Li, M., Knyaz, C., Tamura, K. (2018). MEGA X: Molecular evolutionary genetics analyses across computing platforms. Mol. Biol. Evol. 35: 1547-1549, doi: 10.1093/molbev/msy096
• Kurtzman, C.P., Robnett, C.J. (1998). Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences. Anton. Leeuw. Int. J. G. 73(4): 331-371, doi: 10.1023/A:1001761008817
• Landell, M.F., Hartfelder, C., Valente, P. (2006). Identification and enzymatic profile of yeasts isolated from artisanal cheese in Southern Brazil. Acta Sci. Vet. 34 (1): 49-55.
• Lane, M.M., Morrissey, J.P. (2010). Kluyveromyces marxianus: a yeast emerging from its sister's shadow. Fungal Biol.Rev. 24(1-2): 17-26, doi: 10.1016/j.fbr.2010.01.001
• Lavoie, K., Touchette, M., St-Gelais, D., Labrie, S. (2012). Characterization of the fungal microflora in raw milk and specialty cheeses of the province of Quebec. Dairy Sci. Technol. 92(5): 455-468, doi: 10.1007/s13594-011-0051-4
• Lhomme, E., Lattanzi, A., Dousset, X., Minervini, F., De Angelis, M., Lacaze, G., Onno, B., Gobbetti, M. 2015. Lactic acid bacterium and yeast microbiotas of sixteen French traditional sourdoughs. Int. J. Food Microbiol. 215: 161-170, doi: 10.1016/j.ijfoodmicro.2015.09.015
• Mathara, J. M., Schillinger, U., Kutima, P. M., Mbugua, S. K., Holzapfel, W.H. (2004). Isolation, identification and characterisation of the dominant microorganisms of kule naoto: the Maasai traditional fermented milk in Kenya. Int. J. Food Microbiol. 94(3): 269-278, doi: 10.1016/j.ijfoodmicro.2004.01.008
• McSweeney P.L.H. (2004). Biochemistry of cheese ripening. Int. J. Dairy Technol. 57(2‐3): 127-144, doi: 10.1111/j.1471-0307.2004.00147.x
• Mei, J., Guo, Q., Wu, Y., Li, Y. (2014). Microbial diversity of a Camembert-type cheese using freeze-dried Tibetan kefir coculture as starter culture by culture-dependent and culture-independent methods. PloS one. 9 (10): e111648, doi: 10.1371/journal.pone.0111648
• Moon, S.H., Chang, M., Kim, H.Y., Chang, H.C. (2014). Pichia kudriavzevii is the major yeast involved in film-formation, off-odor production, and texture-softening in over-ripened Kimchi. Food Sci. Biotechnol. 23(2): 489-497, doi: 10.1007/s10068-014-0067-7
• Ocak, E., Javidipour, I. and Y. Tuncturk. (2015). Volatile compounds of Van Herby cheeses produced with raw and pasteurized milks from different species. Int. J. Food Sci. Tech. 52(7): 4315-4323, doi: 10.1007/s13197-014-1458-8
• Ozer, G., Bayraktar, H. (2018). Genetic diversity of Fusarium oxysporum f. sp. cumini isolates analyzed by vegetative compatibility, sequences analysis of the rDNA IGS region and iPBS retrotransposon markers. J. Plant Pathol. 100 (2): 225-232, doi: 10.1007/s42161-018-0063-5
• Padilla, B., Manzanares, P., Belloch, C. (2014). Yeast species and genetic heterogeneity within Debaryomyces hansenii along the ripening process of traditional ewes' and goats' cheeses. Food Microbiol. 38: 160-166, doi: 10.1016/j.fm.2013.09.002
• Psomas, E., Andrighetto, C., Litopoulou-Tzanetaki, E., Lombardi, A., Tzanetakis, N. (2001). Some probiotic properties of yeast isolates from infant faeces and Feta cheese. Int. J. Food Microbiol. 69(1-2): 125-133, doi: 10.1016/S0168-1605(01)00580-3
• Spitaels, F., Wieme, A. D., Janssens, M., Aerts, M., Daniel, H. M., Van Landschoot, A., De Vuyst, L., Vandamme, P. (2014). The microbial diversity of traditional spontaneously fermented lambic beer. PloS one. 9(4): e95384, doi: 10.1371/journal.pone.0095384
• Stepaniak, L. (2004). Dairy enzymology. Int. J. Dairy Technol. 57(2‐3): 153-171, doi: 10.1111/j.1471-0307.2004.00144.x
• Tamura, K., Nei, M. (1993). Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol. Biol. Evol. 10: 512-526, doi: 10.1093/oxfordjournals.molbev.a040023
• Tarakci, Z., Coskun, H. and Tuncturk, Y. (2004). Some Properties of Fresh and Ripened Herby Cheese, a Traditional Variety Produced in Turkey. Food Technol. Biotechno. 42(1): 47-50.
• Thompson, J. D., Higgins, D. G., Gibson, T.J. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22: 4673-4680, doi: 10.1093/nar/22.22.4673.
• Zeng, X., Xia, W., Wang, J., Jiang, Q., Xu, Y., Qiu, Y., Wang, H. (2014). Technological properties of Lactobacillus plantarum strains isolated from Chinese traditional low salt fermented whole fish. Food Control. 40: 351-358, doi: 10.1016/j.foodcont.2013.11.048.
• Zheng, X., Li, K., Shi, X., Ni, Y., Li, B., Zhuge, B. (2018). Potential characterization of yeasts isolated from Kazak artisanal cheese to produce flavoring compounds. Microbiology Open 7(1): e00533, doi: 10.1002/mbo3.533.