Probiyotik kültür immobilizasyonu: stabilite ve optimizasyon çalışmaları
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Date
2019
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Publisher
Trakya Üniversitesi, Fen Bilimleri Enstitüsü
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info:eu-repo/semantics/openAccess
Abstract
Bu tez çalışmasında probiyotik bir bakteri olan Lactobacillus rhamnosus’un immobilize edilmesi, immobilizasyon şartlarının optimizasyonu, farklı koşullarda bakterilerin hayatta kalabilme yeteneklerinin araştırılması amaçlanmıştır. Probiyotik bakterilerin immobilize edilmelerindeki en önemli nedenler; eklendiği gıdadaki canlı kaldığı süreyi uzatmak ve bağırsağa maksimum sayıda mikroorganizma geçebilmesini sağlayabilmektir. Lactobacillus rhamnosus’un immobilize edildiği bu çalışmada sodyum alginat, karragenan ve pektin farklı destek materyalleri olarak denenmiştir. İmmobilizasyonun optimize edilmesi çalışmalarında ise alginat ve CaCl2 yüzdesinin, yüklenen mikroorganizma sayısının etkileri araştırılmıştır. İmmobilizasyonda elde edilen boncuklarda tutulan canlı probiyotik mikroorganizma sayısını belirlemek üzere yapılacak çalışmalar için boncuk patlatma yöntemlerinin (vortex, stomacher, homojenizatör, çalkalamalı inkübatör) etkisi de araştırılmıştır. Ayrıca immobilize bakterinin çeşitli pH değerlerine duyarlılığı, mide asidi ve safraya direnci; serbest haldeki bakteriler kontrol olarak kullanılarak saptanmıştır. Yapılan deneyler sonucunda, en uygun destek materyalinin immobilizasyon etkinliği %89.76 olan sodyum alginat olduğu belirlernmiştir. Optimum parametreler bakteri sayısı 7.46 log CFU/g ile %2’lik alginat, bakteri sayısı 7.47 log CFU/g ile %3’lük CaCl2 ve %25’lik bakteri yüzdesi şeçilmiştir. Mikroorganizmaların boncuklardan çıkarılmasında en etkili yöntemin 7.46 log CFU/g koloni sayısı ile ultrasonik homojenizatör ile patlatma olduğu belirlenmiştir. Serbest haldeki bakterilerin mide asidi ve safra ile muamele edildikten sonra hiç üreyemedikleri, immobilize bakterilerin yapay mide suyunda %68.39, safra tuzunda %58.82 oranında hayatta kaldıkları gözlenmiştir. Sonuç olarak yapılan bu çalışma ile L.rhamnosus’un alginata immobilizasyonu yüksek etkinlik ile gerçekleştirilmiş, immobilizasyon koşulları optimize edilmiş ve immobilizasyonun bakterilerin hayatta kalabilirliği üzerine etkili olduğu saptanmıştır.
In this study, it is aimed to immobilize probiotic bacteria Lactobacillus rhamnosus, to optimize immobilization conditions, to investigate the survival ability of bacteria under different conditions. The most important reasons for the immobilization of probiotic bacteria are extending the period of the food in which it is added and allowing the maximum number of microorganisms passing through the intestine. In this study in which Lactobacillus rhamnosus was immobilized, sodium alginate, carrageenan and pectin were tested as different supportive materials. In the optimization studies for immobilization, the impacts of percentage of alginate and CaCl2 and the number of microorganisms loaded were investigated. The effect of bead blasting methods (vortex, stomacher, homogenizer, shaking incubator) on the studies which determine the number of viable probiotic microorganisms retained in beads obtained from immobilization was investigated. In addition, susceptibility of immobilized bacteria to various pH values, resistance to gastric acid and bile were detected by using free bacteria as a control. As a result of the experiments, it was determined that the most suitable supportive material was sodium alginate which had an immobilization efficiency of 89.76%. The optimum parameters were found to be 2% alginate with 7.46 log CFU / g, 3% CaCl2 and 25% bacteria with 7.47 log CFU / g. The most effective method for removing microorganisms from beads was detonation with an ultrasonic homogenizer with a colony number of 7.46 log CFU / g. It was observed that free bacteria could not survived after treatment with gastric acid and bile, whereas immobilized bacteria survived 68.39% in artificial gastric juice and 58.82% in bile salt. As a result of this study, immobilization of L. rhamnosus by alginate was performed with high efficiency, the conditions of the immobilization were optimized and the immobilization was found to be effective on the viability of the bacteria.
In this study, it is aimed to immobilize probiotic bacteria Lactobacillus rhamnosus, to optimize immobilization conditions, to investigate the survival ability of bacteria under different conditions. The most important reasons for the immobilization of probiotic bacteria are extending the period of the food in which it is added and allowing the maximum number of microorganisms passing through the intestine. In this study in which Lactobacillus rhamnosus was immobilized, sodium alginate, carrageenan and pectin were tested as different supportive materials. In the optimization studies for immobilization, the impacts of percentage of alginate and CaCl2 and the number of microorganisms loaded were investigated. The effect of bead blasting methods (vortex, stomacher, homogenizer, shaking incubator) on the studies which determine the number of viable probiotic microorganisms retained in beads obtained from immobilization was investigated. In addition, susceptibility of immobilized bacteria to various pH values, resistance to gastric acid and bile were detected by using free bacteria as a control. As a result of the experiments, it was determined that the most suitable supportive material was sodium alginate which had an immobilization efficiency of 89.76%. The optimum parameters were found to be 2% alginate with 7.46 log CFU / g, 3% CaCl2 and 25% bacteria with 7.47 log CFU / g. The most effective method for removing microorganisms from beads was detonation with an ultrasonic homogenizer with a colony number of 7.46 log CFU / g. It was observed that free bacteria could not survived after treatment with gastric acid and bile, whereas immobilized bacteria survived 68.39% in artificial gastric juice and 58.82% in bile salt. As a result of this study, immobilization of L. rhamnosus by alginate was performed with high efficiency, the conditions of the immobilization were optimized and the immobilization was found to be effective on the viability of the bacteria.
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Keywords
Lactobacillus Rhamnonus, Probiyotik, İmmobilizasyon, Optimizasyon, Beslenme, Lactobacillus Rhamnonus, Probiotic, İmmobilization, Optimizatio., Nutrition
