Kefiran Ekstraktının Bazı Bitki Patojeni Bakterilerine Karşı Antimikrobiyal Etkisinin Değerlendirilmesi

Kefir; laktik asit bakterileri, asetik asit bakterileri ve mayalar gibi çok çeşitli mikroorganizmaları barındıran granüller tarafından üretilen, pek çok ülkede geleneksel olarak tüketilen fermente bir süt içeceğidir. Granülleri çevreleyen ve ağırlıklı olarak kefiran molekülünden oluşan polisakkarit yapının sağlığı teşvik edici birçok etkisinin yanında çeşitli patojenlere karşı antimikrobiyal etkiye sahip olduğu bildirilmektedir. Bu çalışmada, biri ticari olarak satılan, diğeri yerel bir satıcıdan temin edilen granüller kullanılarak 24 saatlik fermantasyonla kefir üretimi yapılmıştır. Olgunlaşan kefir tanelerinden kefiran ekstraktı eldesi ve saflaştırılması etanol presipitasyon yötemi ile 80°C, 90°C ve 100°C olmak üzere üç farklı sıcaklıkta gerçekleştirilmiştir. Ayrıca elde edilen ekstrakt solüsyonlarının protein içerikleri Bradford yöntemiyle ile belirlenmiştir. Denemelerde kullanılan ekstrakt solüsyonlarının protein içeriği 0,001 g/mL olarak ölçülmüştür. Oluşturulan stok solüsyonlarından, 0,05, 0,1, 1 ve 2 mg ekstrakt içeren uygulamaların Pseudomonas, Xanthomonas, Erwinia ve Clavibacter cinslerine ait bazı bitki patojeni bakterilere karşı antibakteriyel etkisi in vitro koşullarda ilk defa incelenmiştir. Bu amaçla disk difüzyon metodu ve yayma yöntemi olmak üzere iki metot kullanılmıştır. Denemelerde pozitif kontrol olarak kullanılan AN kefir süpernatantı izolatlar üzerinde ortalama 13-17 mm inhibisyon zonu gösterirken, SD süpernatantı 10-14 mm zon üretmiş ancak kefiran ekstralarının antibakteriyel etkisi gözlemlenmemiştir.

Evaluation of the Antimicrobial Effect of Kefiran Extract against Some Plant Pathogenic Bacteria

Kefir; is a fermented milk product which is produced by granules containing a wide variety of microorganisms such as lactic acid bacteria, acetic acid bacteria and yeasts. It is traditionally consumed in many countries. It has been shown in many studies that the polysaccharide structure surrounding the granules which is composed mainly of kefiran molecule has antimicrobial effect against various pathogens as well as many health promoting effects. In this study, 24 h fermented kefir was used with two types of kefir granules for production of kefiran extract. One of them is being sold commercially and the other was collected from private households in a different region of Turkey. Kefiran extraction was carried out from matured kefir granules using three different temperatures, 80°C, 90°C and 100°C. Also, the protein contents of the extracted solutions were determined by Bradford method. Protein content of the extract solutions obtained were measured as 0.001 g/ml. The antibacterial effect of 0.05, 0.1, 1 and 2 mg of this extract against several plant pathogenic bacterial strains belonging to genus Pseudomonas, Xanthomonas, Erwinia ve Clavibacter was investigated in vitro for the first time. For this purpose, two methods, disc diffusion method and spreading method were used. The AN and SD kefir supernatants used as the positive controls in the experiments showed an average of 13-17 mm and 10-14 mm inhibition zones on the isolates, respectively, but the antibacterial effect of kefiran extracts was not observed

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Agrios G. 2005. Plant Pathology. Fifty Ed. Ed.sl.: Elsiver Academic Pres, 922p.
Barbosa AF, Santos PG, Lucho AMS, Schneedorf JM. 2011. Kefiran can disrupt the cell membrane through induced pore formation. J. Electroanal. Chem. 653(1 -2): 61 -66.
Basim E, Basim H, Özcan M. 2006. Antibacterial activities of Turkish pollen and propolis extracts against plant bacterial pathogens. J. Food Eng. 77(4): 992-996.
Bhagwat MK, Datar AG. 2014. Antibacterial activity of herbal extracts against five plant pathogenic bacteria. Arch Phytopathology Plant Protect, 47(7): 892-899.
Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 72(1 -2): 248-254.
Bull CT, De Boer SH, Denny TP, Firrao G, Saux MFL, Saddler GS, Scortichini M, Stead DE, Takikawa, Y. 2010.
Comprehensive list of names of plant pathogenic bacteria, 1980-2007. J Plant Pathol. 551 -592.
Förster H, McGhee GC, Sundin GW, Adaskaveg JE. 2015. Characterization of streptomycin resistance in isolates of Erwinia amylovora from California. Phytopathology 105, 1302–1310.
Furuno T, Nakanishi M. 2012. Kefiran Suppresses AntigenInduced Mast Cell Activation. Biol. Pharm. Bull. 35(2): 178- 183.
Garrote GL, Abraham AG, De Antoni GL. 2001. Chemical and microbiological characterisation of kefir grains. J Dairy Res. 68(4): 639-652.
Güzel-Seydim Z, Wyffels JT, Seydim AC, Greene AK. 2005. Turkish kefir and kefir grains: microbial enumeration and electron microscobic observation. Int. J. Dairy Tech.,58(1): 25-29.
Guzel-Seydim ZB, Kok-Tas T, Greene AK, Seydim AC. 2011. Review: functional properties of kefir. Crit Rev Food Sci Nutr. 51(3): 261 -8.
Guzel-Seydim ZB, Seydim AC, Greene AK, Tas T. 2006. Determination ofantimutagenic properties ofsome fermented milks including changes in the total fatty acid profiles including CLA. Int J Dairy Technol. 59: 209–215.
Ismaiel AA, Ghaly MF, El-Naggar AK. 2011. Milk kefir: ultrastructure, antimicrobial activity and efficacy on aflatoxin B1 production by Aspergillus flavus. Curr Microbiol. 62(5): 602-1609.
Kannan V, Bastas K. 2015. Sustainable Approaches to Controlling Plant Pathogenic Bacteria. ISBN 9780367377441. Boca Raton, Taylor & Francis Ltd. CRC Press.
Liu JR, Chen MJ, Lin CW. 2005. Antimutagenic and antioxidant properties of milk-kefir and soymilk-kefir. J Agric Food Chem.53: 2467–2474.
Liu JR, Wang SY, Chen MJ, Chen HL, Yueh PY, Lin CW. 2006. Hypocholesterolaemc effects of milk-kefir vs soyamilk-kefir in cholesterol-fed hamsters. Br J Nutr. 95: 939–946.
Medrano M, Pérez PF, Abraham AG. 2008. Kefiran antagonizes cytopathic effects ofBacillus cereus extracellular factors. Int. J. Food Microbiol, 122: 1–2.
Micheli L, Uccelletti D, Palleschi C, Crescenzi V. 1999. Isolation and characterisation of a ropy Lactobacillus strain producing the exopolysaccharide kefiran. Appl. Microbiol. Biotechnol.53: 69-74.
Montesanto S, Calò G, Cruciata M, Settanni L, Brucato V, La Carrubba V. 2016. Optimization ofenvironmental conditions for kefiran production by Kefir grain as scaffold for tissue engineering. Chem. Eng. Trans. 49: 607-612
Moradi Z, Kalanpour N. 2019. Kefiran, a branched polysaccharide: Preparation, properties and applications: A review. Carbohydr. Polym. 115100.
Motedayen AA, Khodaiyan F, Salehi EA. 2013. Development and characterisation of composite films made of kefiran and starch. Food Chem. 136(3-4): 1231 -1238.
NCCLS. 2002. National Committee of Clinical Laboratory Standards, approved standard M2-A6. 6thed. Wayne, PA:National Committee for Clinical Lab. Standards. 2002.
Piermaria JA, Pinotti A, Garcia MA, Abraham AG. 2009. Films based on kefiran, an exopolysaccharide obtained from kefir grain: Development and characterization. Food Hydrocoll. 23(3): 684-690.
Pop CR, Salanta L, Rotar AM, Semeniuc CA, Socaciu C, SindicM. 2016. Influence of extraction conditions on characteristics of microbial polysaccharide kefiran isolated from kefir grains biomass. J Food Nutr Res. 55(2).
Rodrigues KL, Caputo LRG, Carvalho JC, Evangelista J, Schneedorf JM. 2005. Antimicrobial and healing activity of kefir and kefiran extract. Int. J. Antimicrob. Agents., 25: 404- 408.
Shiomi M, Sasaki K, Murofushi M, Aibara K. 1982. Antitumor activity in mice orally administered polysaccharide from kefir grain. Jpn. J. Med. Sci. Biol. 35: 75–80.
Sundin GW, Wang N. 2018. Antibiotic Resistance in PlantPathogenic Bacteria. Annu Rev Phytopathol. 56: 161 -180.
Sundin GW, Castiblanco LF, Yuan X, Zeng Q, Yang CH. 2016. Bacterial disease management: challenges, experience, innovation and future prospects: challenges in bacterial molecular plant pathology. Mol Plant Pathol. 17(9): 1506- 1518.
Taşkın B, Akköprü A. 2020. Antibacterial Activity of Different Kefir Types Against Various Plant Pathogenic Bacteria. Journal of Agricultural Sciences. In Press.
Uchida M, Ishii I, Inoue C, Akisato Y, Watanabe K, Hosoyama S, Toida T, Ariyoshi N, Kitada M. 2010. Kefiran reduces atherosclerosis in rabbits fed a high cholesterol diet. J. Atheroscler. Thromb. 17: 980—988.
Vijayendra SVN, Shamala TR. 2014. Film forming microbial biopolymers for commercial applications—A review. Crit Rev Biotechnol. 34(4): 338-357