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Amino Acid Decarboxylase Activity of Some Lactic Acid Bacteria

Year 2015, Volume: 6 Issue: 2, 89 - 94, 31.12.2015

Abstract

Dekarboksilaz aktivitesine sahip olan mikroorganizmalar gıdalarda bulunan amino asitleri enzimatik olarak dekarboksile ederek biyojen amin oluştururlar. Gıdalardaki tipik histamin zehirlenmelerinde yüksek düzeyi en çok peynir ve balık içermektedir. Bu nedenle fermente ürünlerde kültür seçiminde dekarboksilaz aktivitesi önemli bir problemdir. Bu çalışmada, 20 adet peynir örneğinden izole edilen 161 laktik asit bakterisinin dekarboksilaz aktivitesi belirlenmiştir. Çalışmanın sonucunda, 59 Enterococcus spp.’nin 37’si (62.71%), 47 Lactococcus spp.’nin 4’ü (8.51%), 55 Lactobacillus spp.’nin 4’ü (7.27%) tirosinden tiramin ve 55 Lactobacillus spp.’nin 3 ‘ü (5.45%) tiriptofandan triptamin üretmiştir. 108 suşun (70.19%) hiçbirinde ornitin, histidin, tirozin, tiriptofan, fenilalanin, ya da lizin dekarboksile edememiştir. Bu izolatların peynir endüstrisi için uygun başlatıcı kültür olduğu düşünülmektedir.

Anahtar Kelimeler: Dekarboksilaz aktivitesi, laktik asit bakterileri, peynir

References

  • Ayhan, K., Alsancak-Ökan, G., Noveir, MR. (2000). Determi- nation of Histidine Decarboxylase Activity Produced by Enterobacteriaceae İsolated from Ground Meat. Ciencia, 8 (2): 243-250.
  • Bover-Cid, S., Holzapfel, W.H. (1999). Improve Screening Procedure for Biogenic Amine Production by Lactic Acid Bacteria. International Journal of Food Microbiology, 53: 33-41.
  • Buñková, L., Buñka, F., Mantlová, G., Ĉablová, A., Sedláĉek, I., Švec, P., Pachlová, V., Kráĉmar, S. (2010). The Effect of Ripening and Storage Conditions on the Distribution of Tyramine, Putrescine and Cadaverine in Edam-Cheese. Food Microbiology. 27: 880- 888.
  • Buňková, L., Buñka, F., Pollaková, E., Podešvová, T., Dráb, V. (2011). The Effect of Lactose, Nacl and an Ae- ro/Anaerobic Environment on the Tyrosine Decarboxylase Activity of Lactococcus Lactis Subsp. Cremoris and Lac- tococcus Lactis Subsp. Lactis. International Journal of Food Microbiology. 147: 112-119.
  • Buňková, L., Adamcová, G., Hudcová, K., Velichová, H., Pachlová, V., Lorencová, E., Buñka, F. (2013). Monitoring of Biogenic Amines in Cheeses Manufactured at Small- Scale Farms and in Fermented Dairy Products in the Czech Republic. Food Chemistry. 141: 548-551.
  • Celano, GV., Cafarchia, C., Buja, F., Tiecco, G., (1992). Ri- cerca Di Amine Biogene in Alcuni Formaggi. Ind Aliment, 31: 764-768.
  • Christensen, J.E., Dudley, E.G., Pederson, J.A., Steele, J.L. (1999). Peptidases and Amino Acid Catabolism in Lactic Acid Bacteria. Antonie Van Leeuwenhoek 76, 217 - 246.
  • Durlu-Özkaya, F., Alichanidis, E., Litopoulou-Tzanetaki, E., Tunail, N. (1999). Determination of Biogenic Amine Con- tent of Beyaz Cheese and Biogenic Amine Production Ability of Some Lactic Acid Bacteria. Milchwissen, 54 (12): 680-682.
  • Gingerich, T.M., Lorca, T., Flick, G.J., Pierson, M.D., Mc Nair, H.M. (1999). Biogenic Amine Survey and Organoleptic Changes in Fresh, Stored and Temperature-Abused Blue- fish. Journal Food Protection, 62: 1033-1037.
  • Giraffa, G., Pepe, G., Locci, F., Neviani, E., Carminati, D. (1995). Hemolytic Activity, Production of Thermonuclease and Biogenic Amines by Dairy Enterococci. Italian Journal of Food Science, 4: 341-349.
  • Halász, A., Baráth, Á., Simon-Sarkadi, L., Holzapfel, W. (1994). Biogenic Amines and Their Production by Micro- organisms in Food. Trends in Food Science & Techno- logy, 5: 42-49.
  • Joosten HMLJ Northolt, MD. (1989). Detection, Growth, and Amine-Producing Capacity of Lactobacilli in Cheese. Appl Environ Microbiol, 55 (9): 2356-2359.
  • Kalaĉ, P., Ŝpiĉka, J., Křižek, M., Stediedlová, Ŝ., Peikánová, T., (1999). Concentrations of Seven Biogenic Amines in Sauerkraut. Food Chemistry, 67: 275-280.
  • Kalhotka, L., Cwiková, O., Čírtková (Kovářová), V., Ma- toušovál, Z., Přichystalová, J. (2012). Changes in Counts of Microorganisms and Biogenic Amines Production Du- ring the Manufacture of Fermented Sausages Poličan. Journal of Microbiology, Biotechnology and Food Scien- ces, 2(2): 667-683.
  • Karmas, E. (1981). Biogenic Amines as İndicators of Seafood Freshness. Lebensm. Wiss. Technol. 14: 273–275.
  • Komprda, T., Burdychová, R., Dohnal, V., Cwiková, O., Slád- ková, P., DvoRáCková, H. (2008). Tyramine Production in Dutch-Type Semi-Hard Cheese from Two Different Pro- ducers. Food Microbiol. 25: 219-227.
  • Kucerová, K., Svobodová, H., Tûma, S., Ondrácková, I., Plocková, M. (2009). Production of Biogenic Amines by Enterococci. Czech J Food Sci, 27 (2): 50-55.
  • Landete, J. M., Arena, M. E., Pardo, I., Manca De Nadra, M. C., Ferrer, S. (2008). Comparative Survey of Putrescine Production from Agmatine Deamination in Different Bacte- ria. Food Microbiology, 25: 882–887
  • Ladero, V., Fernández, M., Cuesta, I., Alvarez, M.A. (2010). Quantitative Detection and İdentification of Tyramine- Producing Enterococci and Lactobacilli in Cheese by Mul- tiplex Qpcr. Food Microbiology. 27, 933-939.
  • Ladero, V., Canedo, E., Perez, M., Martin, M. C., Fernandez, M., Alvarez, M. A. (2012). Multiplex Qpcr for the Detec- tion and Quantification of Putrescine Producing Lactic Acid Bacteria in Dairy Products. Food Control, 27: 307– 313.
  • Marcobal, A., De Las Rivas, B., García-Moreno, E., Muñoz, R. (2004). The Tyrosine Decarboxylation Test does not Differentiate Enterococcus Faecalis from Enterococcus Faecium. Syst. Appl. Microbiol. 2: 423 - 426.
  • Marcobal, A., De Las Rivas, B., Muñoz, R. (2006). First Ge- netic Characterization of a Bacterial Β-Phenylethylamine Biosynthetic Enzyme in Enterococcus Faecium RM58. FEMS Microbiology Letters 258: 144–149.
  • Novella-Rodríguez, S., Veciana-Nogués, M.T., Roig-Sagués, A.J., Trujillo-Mesa, A.J., Vidal-Carou, M.C. (2002). Influ- ence of Starter and Non-Starter on the Formation of Bio- genic Amine in Goat Cheese During Ripening. J. Dairy Sci. 85: 2471-2478.
  • Paulsen, P., Bauer, F., Vali, S. (1997). Biogenic Amines in Fermented Sausage. 1. Methods for the Determination of Biogenic Amines. Fleischwirtsch, 77: 450-452.
  • Rea, M.C., Franz, C.M., Holzapfel, W.H., Cogan, T.M. (2004). Development of Enterococci and Production of Tyramine During the Manufacture and Ripening of Cheddar Chee- se. Irish J. Agricult. Food Res. 43: 247-258.
  • Roig-Sagués, A.X., Hernández-Herrero, M.M., López- Sabater, E.I., Rodríguez-Jerez, J. J., Mora-Ventura, M.T. (1999). Microbiological Events During the Elaboration of “Fuet”, A Spanish Ripened Sausage. Relationships Between the Development of Histidine and Tyrosine De- carboxylase Containing Bacteria and Ph and Water Acti- vity. Eur. Food Res. Technol. 209: 108-112.
  • Santos, S. (1996). Biogenic Amines: Their Importance in Fo- ods. International Journal of Food Microbiology, 29: 213- 231.
  • Sarantinopoulos, P., Andrighetto, C., Georgalaki, M.D., Rea, M.C., Lombardi, A., Cogan, T.M., Kalantzopoulos, G., Tsakalidou, E. (2001). Biochemical Properties of Entero- cocci Relevant to their Technological Performance. Inter- national Dairy Journal, 11(8): 621-647.
  • Schirone M, Tofalo R, Visciano P, Corsetti A, Suzzi G. (2012). Biogenic Amines in Italian Pecorino Cheese. Frontiers in Microbiol, NCBI, 3, 171.
  • Shalaby, A.R. (1996). Significance of Biogenic Amines to Food Safety and Human Health. Food Research Interna- tional, 29(7): 675-690.
  • Santos, M. H. (1996). Biogenic Amines: Their İmportance in Food. International Journal of Food Microbiology, 29: 213–231.
  • Sinell, H.J. (1978). Biogene Amine Als Risiko-Faktoren in Der Fischhygiene. Archiv Fur Lebensmittel-Hygiene, 29: 206- 210.
  • Smit, G., Smit, B.A., Engels, W.J.M. (2005). Flavour Forma- tion by Lactic Acid Bacteria and Biochemical Flavour Pro- filing of Cheese Products. FEMS Microbiol. Rev. 29: 591 - 610.
  • Sumner, SS., Taylor, SL. (1989). Detection Method for His- tamine-Producing, Dairy-Related Bacteria Using Diamine Oxidase Leucocrystal Violet. J of Food Protect, 52 (2): 105-108.
  • Ten Brink, B., Damink, C., Joosten, H.M.L.J., Huis İn’t Veld, J.H.J. (1990). Occurance and Formation of Biological Ac- tive Amines in Foods. International Journal of Food Mic- robiology, 11(1): 73-84.
  • Tunail, N., Özkaya, FD., Gürsel, A., Tamuçay, B. (2001) Star- ter Bakterilerin Oluşturdukları Biyojen Aminlerin Saptan- ması ve Salamura Beyaz Peynirdeki Biyojen Amine Bazı Risk Faktörünün Belirlenmesi. Ankara Üniversitesi Araş- tırma Fonu, Proje No: 96-11-12-04, Ankara.
  • Tuncer, Y. (2009). Some Technological Properties of Phe- notypically İdentified Enterococci Strains İsolated from Turkish Tulum Cheese. African J Of Biotech, 8 (24): 7008- 7016.
  • Turantaş, F., Öksüz, A. (1998). Balık ve Balık Ürünlerinde Biyojenik Aminler ve Amin Üretiminde Rol Oynayan Bak- teriler. Gıda Teknolojisi. 3(5): 58-65.
  • Valsamaki, K., Michaelidou, A., Polychroniadou, A. (2000). Biogenic Amine Production in Feta Cheese. Food Chem. 71: 259 - 266.
  • Vidal-Carou, M.C., Izquierdo-Pulido, M., Martın-Morro, M., Marine-Font, A. (1990). Histamine and Tyramine İn Meat Products: Relationship with Meat Spoilage. Food Chem. 37: 239–349.
  • Yerlikaya, P., Gökoğlu, N. (2002). Gıdalarda Biyojen Aminler ve Önemi. Gıda Mühendisliği Dergisi, 12(6): 24-30.
  • Yousif, N.M.K., Dawyndt, P., Abriouel, H., Wijaya, A., Schil- linger, U., Vancanneyt, M., Swings, J., Dirar, H.A., Hol- zapfel, W.H., Franz, C.M.A.P. (2005). Molecular Charac- terization, Technological Properties and Safety Aspects of Enterococci from ‘Hussuwa’, an African Fermented Sorghum Product. Journal of Applied Mirobiology, 96: 216-228.

Amino Acid Decarboxylase Activity of Some Lactic Acid Bacteria

Year 2015, Volume: 6 Issue: 2, 89 - 94, 31.12.2015

Abstract

Dekarboksilaz aktivitesine sahip olan mikroorganizmalar gıdalarda bulunan amino asitleri enzimatik olarak dekarboksile ederek biyojen amin oluştururlar. Gıdalardaki tipik histamin zehirlenmelerinde yüksek düzeyi en çok peynir ve balık içermektedir. Bu nedenle fermente ürünlerde kültür seçiminde dekarboksilaz aktivitesi önemli bir problemdir. Bu çalışmada, 20 adet peynir örneğinden izole edilen 161 laktik asit bakterisinin dekarboksilaz aktivitesi belirlenmiştir. Çalışmanın sonucunda, 59 Enterococcus spp.’nin 37’si (62.71%), 47 Lactococcus spp.’nin 4’ü (8.51%), 55 Lactobacillus spp.’nin 4’ü (7.27%) tirosinden tiramin ve 55 Lactobacillus spp.’nin 3 ‘ü (5.45%) tiriptofandan triptamin üretmiştir. 108 suşun (70.19%) hiçbirinde ornitin, histidin, tirozin, tiriptofan, fenilalanin, ya da lizin dekarboksile edememiştir. Bu izolatların peynir endüstrisi için uygun başlatıcı kültür olduğu düşünülmektedir

References

  • Ayhan, K., Alsancak-Ökan, G., Noveir, MR. (2000). Determi- nation of Histidine Decarboxylase Activity Produced by Enterobacteriaceae İsolated from Ground Meat. Ciencia, 8 (2): 243-250.
  • Bover-Cid, S., Holzapfel, W.H. (1999). Improve Screening Procedure for Biogenic Amine Production by Lactic Acid Bacteria. International Journal of Food Microbiology, 53: 33-41.
  • Buñková, L., Buñka, F., Mantlová, G., Ĉablová, A., Sedláĉek, I., Švec, P., Pachlová, V., Kráĉmar, S. (2010). The Effect of Ripening and Storage Conditions on the Distribution of Tyramine, Putrescine and Cadaverine in Edam-Cheese. Food Microbiology. 27: 880- 888.
  • Buňková, L., Buñka, F., Pollaková, E., Podešvová, T., Dráb, V. (2011). The Effect of Lactose, Nacl and an Ae- ro/Anaerobic Environment on the Tyrosine Decarboxylase Activity of Lactococcus Lactis Subsp. Cremoris and Lac- tococcus Lactis Subsp. Lactis. International Journal of Food Microbiology. 147: 112-119.
  • Buňková, L., Adamcová, G., Hudcová, K., Velichová, H., Pachlová, V., Lorencová, E., Buñka, F. (2013). Monitoring of Biogenic Amines in Cheeses Manufactured at Small- Scale Farms and in Fermented Dairy Products in the Czech Republic. Food Chemistry. 141: 548-551.
  • Celano, GV., Cafarchia, C., Buja, F., Tiecco, G., (1992). Ri- cerca Di Amine Biogene in Alcuni Formaggi. Ind Aliment, 31: 764-768.
  • Christensen, J.E., Dudley, E.G., Pederson, J.A., Steele, J.L. (1999). Peptidases and Amino Acid Catabolism in Lactic Acid Bacteria. Antonie Van Leeuwenhoek 76, 217 - 246.
  • Durlu-Özkaya, F., Alichanidis, E., Litopoulou-Tzanetaki, E., Tunail, N. (1999). Determination of Biogenic Amine Con- tent of Beyaz Cheese and Biogenic Amine Production Ability of Some Lactic Acid Bacteria. Milchwissen, 54 (12): 680-682.
  • Gingerich, T.M., Lorca, T., Flick, G.J., Pierson, M.D., Mc Nair, H.M. (1999). Biogenic Amine Survey and Organoleptic Changes in Fresh, Stored and Temperature-Abused Blue- fish. Journal Food Protection, 62: 1033-1037.
  • Giraffa, G., Pepe, G., Locci, F., Neviani, E., Carminati, D. (1995). Hemolytic Activity, Production of Thermonuclease and Biogenic Amines by Dairy Enterococci. Italian Journal of Food Science, 4: 341-349.
  • Halász, A., Baráth, Á., Simon-Sarkadi, L., Holzapfel, W. (1994). Biogenic Amines and Their Production by Micro- organisms in Food. Trends in Food Science & Techno- logy, 5: 42-49.
  • Joosten HMLJ Northolt, MD. (1989). Detection, Growth, and Amine-Producing Capacity of Lactobacilli in Cheese. Appl Environ Microbiol, 55 (9): 2356-2359.
  • Kalaĉ, P., Ŝpiĉka, J., Křižek, M., Stediedlová, Ŝ., Peikánová, T., (1999). Concentrations of Seven Biogenic Amines in Sauerkraut. Food Chemistry, 67: 275-280.
  • Kalhotka, L., Cwiková, O., Čírtková (Kovářová), V., Ma- toušovál, Z., Přichystalová, J. (2012). Changes in Counts of Microorganisms and Biogenic Amines Production Du- ring the Manufacture of Fermented Sausages Poličan. Journal of Microbiology, Biotechnology and Food Scien- ces, 2(2): 667-683.
  • Karmas, E. (1981). Biogenic Amines as İndicators of Seafood Freshness. Lebensm. Wiss. Technol. 14: 273–275.
  • Komprda, T., Burdychová, R., Dohnal, V., Cwiková, O., Slád- ková, P., DvoRáCková, H. (2008). Tyramine Production in Dutch-Type Semi-Hard Cheese from Two Different Pro- ducers. Food Microbiol. 25: 219-227.
  • Kucerová, K., Svobodová, H., Tûma, S., Ondrácková, I., Plocková, M. (2009). Production of Biogenic Amines by Enterococci. Czech J Food Sci, 27 (2): 50-55.
  • Landete, J. M., Arena, M. E., Pardo, I., Manca De Nadra, M. C., Ferrer, S. (2008). Comparative Survey of Putrescine Production from Agmatine Deamination in Different Bacte- ria. Food Microbiology, 25: 882–887
  • Ladero, V., Fernández, M., Cuesta, I., Alvarez, M.A. (2010). Quantitative Detection and İdentification of Tyramine- Producing Enterococci and Lactobacilli in Cheese by Mul- tiplex Qpcr. Food Microbiology. 27, 933-939.
  • Ladero, V., Canedo, E., Perez, M., Martin, M. C., Fernandez, M., Alvarez, M. A. (2012). Multiplex Qpcr for the Detec- tion and Quantification of Putrescine Producing Lactic Acid Bacteria in Dairy Products. Food Control, 27: 307– 313.
  • Marcobal, A., De Las Rivas, B., García-Moreno, E., Muñoz, R. (2004). The Tyrosine Decarboxylation Test does not Differentiate Enterococcus Faecalis from Enterococcus Faecium. Syst. Appl. Microbiol. 2: 423 - 426.
  • Marcobal, A., De Las Rivas, B., Muñoz, R. (2006). First Ge- netic Characterization of a Bacterial Β-Phenylethylamine Biosynthetic Enzyme in Enterococcus Faecium RM58. FEMS Microbiology Letters 258: 144–149.
  • Novella-Rodríguez, S., Veciana-Nogués, M.T., Roig-Sagués, A.J., Trujillo-Mesa, A.J., Vidal-Carou, M.C. (2002). Influ- ence of Starter and Non-Starter on the Formation of Bio- genic Amine in Goat Cheese During Ripening. J. Dairy Sci. 85: 2471-2478.
  • Paulsen, P., Bauer, F., Vali, S. (1997). Biogenic Amines in Fermented Sausage. 1. Methods for the Determination of Biogenic Amines. Fleischwirtsch, 77: 450-452.
  • Rea, M.C., Franz, C.M., Holzapfel, W.H., Cogan, T.M. (2004). Development of Enterococci and Production of Tyramine During the Manufacture and Ripening of Cheddar Chee- se. Irish J. Agricult. Food Res. 43: 247-258.
  • Roig-Sagués, A.X., Hernández-Herrero, M.M., López- Sabater, E.I., Rodríguez-Jerez, J. J., Mora-Ventura, M.T. (1999). Microbiological Events During the Elaboration of “Fuet”, A Spanish Ripened Sausage. Relationships Between the Development of Histidine and Tyrosine De- carboxylase Containing Bacteria and Ph and Water Acti- vity. Eur. Food Res. Technol. 209: 108-112.
  • Santos, S. (1996). Biogenic Amines: Their Importance in Fo- ods. International Journal of Food Microbiology, 29: 213- 231.
  • Sarantinopoulos, P., Andrighetto, C., Georgalaki, M.D., Rea, M.C., Lombardi, A., Cogan, T.M., Kalantzopoulos, G., Tsakalidou, E. (2001). Biochemical Properties of Entero- cocci Relevant to their Technological Performance. Inter- national Dairy Journal, 11(8): 621-647.
  • Schirone M, Tofalo R, Visciano P, Corsetti A, Suzzi G. (2012). Biogenic Amines in Italian Pecorino Cheese. Frontiers in Microbiol, NCBI, 3, 171.
  • Shalaby, A.R. (1996). Significance of Biogenic Amines to Food Safety and Human Health. Food Research Interna- tional, 29(7): 675-690.
  • Santos, M. H. (1996). Biogenic Amines: Their İmportance in Food. International Journal of Food Microbiology, 29: 213–231.
  • Sinell, H.J. (1978). Biogene Amine Als Risiko-Faktoren in Der Fischhygiene. Archiv Fur Lebensmittel-Hygiene, 29: 206- 210.
  • Smit, G., Smit, B.A., Engels, W.J.M. (2005). Flavour Forma- tion by Lactic Acid Bacteria and Biochemical Flavour Pro- filing of Cheese Products. FEMS Microbiol. Rev. 29: 591 - 610.
  • Sumner, SS., Taylor, SL. (1989). Detection Method for His- tamine-Producing, Dairy-Related Bacteria Using Diamine Oxidase Leucocrystal Violet. J of Food Protect, 52 (2): 105-108.
  • Ten Brink, B., Damink, C., Joosten, H.M.L.J., Huis İn’t Veld, J.H.J. (1990). Occurance and Formation of Biological Ac- tive Amines in Foods. International Journal of Food Mic- robiology, 11(1): 73-84.
  • Tunail, N., Özkaya, FD., Gürsel, A., Tamuçay, B. (2001) Star- ter Bakterilerin Oluşturdukları Biyojen Aminlerin Saptan- ması ve Salamura Beyaz Peynirdeki Biyojen Amine Bazı Risk Faktörünün Belirlenmesi. Ankara Üniversitesi Araş- tırma Fonu, Proje No: 96-11-12-04, Ankara.
  • Tuncer, Y. (2009). Some Technological Properties of Phe- notypically İdentified Enterococci Strains İsolated from Turkish Tulum Cheese. African J Of Biotech, 8 (24): 7008- 7016.
  • Turantaş, F., Öksüz, A. (1998). Balık ve Balık Ürünlerinde Biyojenik Aminler ve Amin Üretiminde Rol Oynayan Bak- teriler. Gıda Teknolojisi. 3(5): 58-65.
  • Valsamaki, K., Michaelidou, A., Polychroniadou, A. (2000). Biogenic Amine Production in Feta Cheese. Food Chem. 71: 259 - 266.
  • Vidal-Carou, M.C., Izquierdo-Pulido, M., Martın-Morro, M., Marine-Font, A. (1990). Histamine and Tyramine İn Meat Products: Relationship with Meat Spoilage. Food Chem. 37: 239–349.
  • Yerlikaya, P., Gökoğlu, N. (2002). Gıdalarda Biyojen Aminler ve Önemi. Gıda Mühendisliği Dergisi, 12(6): 24-30.
  • Yousif, N.M.K., Dawyndt, P., Abriouel, H., Wijaya, A., Schil- linger, U., Vancanneyt, M., Swings, J., Dirar, H.A., Hol- zapfel, W.H., Franz, C.M.A.P. (2005). Molecular Charac- terization, Technological Properties and Safety Aspects of Enterococci from ‘Hussuwa’, an African Fermented Sorghum Product. Journal of Applied Mirobiology, 96: 216-228.
There are 42 citations in total.

Details

Primary Language English
Journal Section Research Paper
Authors

Pelin Ertürkmen

İlkay Turhan This is me

Zübeyde Öner

Publication Date December 31, 2015
Published in Issue Year 2015 Volume: 6 Issue: 2

Cite

APA Ertürkmen, P., Turhan, İ., & Öner, Z. (2015). Amino Acid Decarboxylase Activity of Some Lactic Acid Bacteria. Mehmet Akif Ersoy Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 6(2), 89-94.