Doğal Fermente Gıdalardan İzole Edilen Muhtemel Laktik Asit Bakterilerinin Antimikrobiyal Aktiviteleri ve Laktik Asit Üretim Düzeylerinin İncelenmesi

Year 2022, Volume: 36 Issue: 1, 25 - 40, 01.06.2022

Özüm Özoğlu Mehmet Gumustas Sibel A. Özkan Evrim Güneş Altuntaş

https://doi.org/10.20479/bursauludagziraat.943244

Abstract

Laktik asit bakterileri pek çok fermente ve probiyotik gıdanın bünyesinde yer alan faydalı bakterilerdir. Bu bakterileri önemli kılan özellikleri arasında GRAS (Generally Recognized As Safe) statüde olmaları ve ürettikleri antimikrobiyal metabolitler bulunmaktadır. Laktik asit bakterilerinin ürettiği antagonistik etkiye sahip en önemli metabolit bu gruba adını veren laktik asit olup; bunun yanısıra hidrojen peroksit, asetik asit, diasetil, bakteriyosin vb. metabolitler ile de antagonistik etkiye neden olmaktadırlar. Bu çalışmada çeşitli peynir, sucuk ve kefir örneklerinden izole edilen 23 adet muhtemel laktik asit bakteri izolatının Salmonella Enteritidis ATCC 13076, Streptococcus mutans ATCC 25175, Escherichia coli ATCC 25922, Bacillus subtilis ATCC 6633 ve Staphylococcus aureus ATCC 43300 (metisilin ve oksasilin dirençli) patojenleri üzerindeki inhibisyon etkisi Agar Spot Yöntemi ile araştırılmıştır. Test sonucunda pozitif etki gösteren izolatların süpernatantları ile Kuyu Difüzyon denemesi gerçekleştirilmiştir. Son olarak izolatların ürettiği laktik asit miktarları HPLC yöntemi ile de belirlenmiştir. Agar Spot Testi’ne göre izolatların neredeyse tamamı tüm patojenler üzerinde etkinlik gösterirken, patojenler üzerindeki inhibisyon etkiyi gösteren zon çaplarının 1-24.5 mm aralığında değiştiği tespit edilmiştir. 24.5 mm zon çapı ile maksimum etki 31 nolu izolat tarafından E.coli ATCC 25922 üzerinde gözlemlenirken; 1 mm zon çapı ile minimum etki 431 nolu izolat tarafından S. Enteritidis ATCC 13076 test mikroorganizmasına karşı gözlemlenmiştir. Kuyu Difüzyon Testi’nde ise hiçbir pozitif sonuç elde edilememiş ve bu nedenle bakterilerin antagonistik etkilerinin daha çok laktik asit üretiminden kaynaklandığı düşünülmüştür. Yapılan kromatografik çalışma ile laktik asit bakterilerinin ürettiği laktik asit miktarının 0.13-5.52 mmol aralığında olduğu gözlenmiştir. Çalışmadan elde edilen sonuçlar laktik asit bakterilerinin bakteriyosin gibi önemli protein bazlı inhibitörleri üretmese bile, ürettiği temel metabolit olan laktik asit ile etkin bir antimikrobiyal aktivite kapasitesine sahip olduğunu göstermektedir.

Keywords

Antimikrobiyal aktivite, laktik asit, laktik asit bakterisi, HPLC

Supporting Institution

Ankara Üniversitesi Bilimsel Araştırmalar Proje Koordinatörlüğü

Project Number

17H0415001

Thanks

Çalışmada kullanılan patojen suşların temininde sağladıkları destek için Ankara Üniversitesi Eczacılık Fakültesi Farmasötik Mikrobiyoloji Anabilim Dalı Öğretim Üyeleri Doç. Dr. Banu KAŞKATEPE ve Doç. Dr. Müjde ERYILMAZ’a teşekkür ederiz.

Investigation of Antimicrobial Activities and Lactic Acid Production Levels of Presumptive Lactic Acid Bacteria Isolated From Naturally Fermented Foods

Abstract

Lactic acid bacteria are useful bacteria which are included to composition of many fermented and probiotic food. Being GRAS (Generally Recognized As Safe) statute and the antimicrobial metabolites produced by them are among the properties which make these bacteria significant. The most important metabolite with antagonistic effect produced by lactic acid bacteria is lactic acid that gives the name of these bacteria, besides LAB can have antagonistic effect by producing hydrogen peroxide, acetic acid, diacetyl, bacteriocin etc. In this study, inhibition effects of 23 presumptive lactic acid bacteria isolates that isolated from various cheese, sucuk and kefir samples on Salmonella Enteritidis ATCC 13076, Streptococcus mutans ATCC 25175, Escherichia coli ATCC 25922, Bacillus subtilis ATCC 6633 and Staphylococcus aureus ATCC 43300 (methicillin and oxacillin resistant) were investigated by Agar Spot method. According to the assay, Well Diffusion Test was performed with supernatant of the isolates showed positive results. Finally, the quantification of lactic acid was detected by HPLC method. As a result of Agar Spot Test, while all of the isolates were effective on all pathogens, the inhibition zone diameters were ranged between 1-24.5 mm. While the maximum effect was observed by isolate 31 on E.coli ATCC 25922 with a zone diameter of 24.5 mm, the minimum effect was observed by isolate 431 against S. Enteritidis ATCC 13076 test microorganism with a zone diameter of 1 mm.Then, due to not to have any positive results from Well Diffusion Assay, it has been considered the antagonistic effect of the bacteria were originated from lactic acid production. It was observed that lactic acid produced by LAB was ranged between 0.13-5.52 mmol by chromatographic study. Even if the LAB do not produce protein-based inhibitors such as bacteriocin, the results shows LAB has an effective antimicrobial activity capacity with lactic acid, the main metabolite of LAB.

Keywords

Antimicrobial activity, lactic acid, lactic acid bacteria, HPLC

Project Number

17H0415001

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