DETERMINATION OF TANNASE ACTIVITY OF THE LACTIC ACID BACTERIA THAT ISOLATED FROM FERMENTED PRODUCTS

Öz

In this study, 10 isolates obtained from fermented products in previous studies were identified byDuPont Qualicon RiboPrinter®system. Tannase activity was determined by spectrophotometric andvisual reading methods. Lactobacillus curvatus P3X showed the highest tannase activity. Approximatemolecular weight of the enzyme was determined by SDS- PAGE method. The effect of differenttemperatures, pH values, substrate concentrations, and the concentrations of tannic acid and variousminerals, on activity of tannase of L. curvatus P3X, of which showed high tannase activity, wereinvestigated. As a result of tests, the optimum temperature was found as 37°C, optimum pH was foundas 5.0, methyl gallate concentration that was used as the substrate was found as 7 mM and tannic acidconcentration was found as 1.75 mM. Tannase activity of lactic acid bacteria decreased in the presenceof Hg+2and Mg+2ions, Ca+2and Zn+2ions did not show any effect on tannase activity. In addition, it wasdetected that surfactant (Tween 80), chelate (EDTA), inhibitor (DMSO), and denaturing agents (urea)had no effect on tannase activity

Anahtar Kelimeler

• Tannase, lactic acid bacteria, methyl gallate

FERMENTE GIDALARDAN İZOLE EDİLEN LAKTİK ASİT BAKTERİLERİNİN TANNAZ AKTİVİTESİNİN BELİRLENMESİ

Öz

Bu çalışmada, daha önceki çalışmalarda fermente ürünlerden izole edilen 10 izolatın DuPont QualiconRiboPrinter®sistem ile tanımlamaları yapılmıştır. Spektrofotometrik ve görsel okuma metodu ile tannazaktivitesi belirlenmiştir. En yüksek aktiviteyi Lactobacillus curvatus P3X izolatı göstermiştir. L. curvatusP3X’e ait enzimin yaklaşık molekül ağırlığı SDS-PAGE yöntemi ile belirlenmiştir. Yüksek tannaz aktivitesigösteren L. curvatus P3X’in tannaz aktivitesi üzerine farklı sıcaklık, pH, substrat yoğunluğu, tannik asitkonsantrasyonu ve çeşitli minerallerin etkisi araştırılmıştır. Yapılan testler sonucunda optimum sıcaklık37°C, pH 5.0 substrat olarak kullanılan metil gallat konsantrasyonu 7 mM, tannik asit konsantrasyonuise 1.75 mM olarak bulunmuştur. Laktik asit bakterilerinin tannaz aktivitesi Hg+2ve Mg+2iyonlarınınvarlığında düşerken, Ca+2ve Zn+2’nun tannaz aktivitesi üzerine etki yapmadığı görülmüştür. Ayrıcayüzey aktif madde (Tween 80), şelat (EDTA), inhibitör (DMSO) ve yapıyı bozan maddenin de (Üre)tannaz aktivitesine tesir etmediği saptanmıştır

Anahtar Kelimeler

• Tannaz, laktik asit bakterisi, metil galat

Kaynakça

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