Yasemin KILIÇ, Zehra Nur YÜKSEKDAĞ, Hazer YÜKSEKDAĞ

Lactobacillus ve Bifidobacterium Cinsi Bakterilerin Beta Galaktosidaz Enzim Aktivitelerinin Belirlenmesi

Bu çalışmada, insan, gıda ve hayvan kaynaklı 39 Lactobacilllus cinsine ait ve yeni doğan gaitasından izole edilmiş 3 Bifidobacterium cinsine ait toplam 42 bakteri kullanılmıştır. O-nitrofenil-beta-D-galaktosit (o-NPG) substrat olarak kullanılarak, kültürlerin β-galaktosidaz enzim ve spesifik aktiviteleri belirlenmiştir. Lactobacillus cinsine ait kültürlerden L. fermentum ZYN17 (2.468 U/mg), L. casei LB65 (1.116 U/mg), L. rhamnosus GD11 (1.034 U/mg) ve L. acidophilus BAZ36 (0.947 U/mg) suşlarının, Bifidobacterium cinsine ait kültürlerden de B. breve A26 (0.726 U/mg) suşunun en yüksek spesifik aktivite yeteneğine sahip oldukları tespit edilmiştir. Ayrıca, bakterilerin 5-brom-4-klor-3-indolil-β-D-galaktopiranosit (X-gal) substrat bileşiğiyle de nitel olarak enzim aktivitesinin varlığı değerlendirilmiştir. Yüksek spesifik β-galaktozidaz aktivitesi gösteren ZYN17 suşuna ait β-galaktozidaz enziminin optimizasyonu yapılmıştır. β-galaktozidaz enziminin optimum pH’sı 6.8, optimum sıcaklığı 37 ºC ve optimum tamponun potasyum fosfat tamponu olduğu belirlenmiştir.

Anahtar Kelimeler:

Lactobacilllus, Bifidobacterium, β-galaktosidaz enzim aktivitesi

Beta Galactosidase Enzyme Activities of Lactobacillus and Bifidobacterium Genus (Turkish with English Abstract)

In this study total 42 bacteria were used; 39 Lactobacilllus originated from human, food and animal and 3 Bifidobacterium isolated from new born baby feces. The β-galactosidase enzyme activities and specific activities were determined by using o-nitrophenyl-b-D-galactopyranoside (o-NPG) as a substrate. The highest specific enzyme activities observed among Lactobacilli cultures were at L. fermentum ZYN17 (2.468 U/mg), L. casei LB65 (1.116 U/mg), L. rhamnosus GD11 (1.034 U/mg), and L. acidophilus BAZ36 (0.947 U/mg) strains and the highest enzyme activities observed among Bifidobacterium was at B. breve A26 (0.726 U/mg) strain. The enzyme activities of bacteria were also determined qualitatively by using the 5-bromo-4-chloro-3-indolyl-β-D-galaktoside (X-gal) substrate compound. The enzyme optimization of ZYN17 strain with highest specific β-galactosidase activity was achieved. The optimum pH was determined as 6.8, optimum temperature was 37 ºC and the optimum buffer was potassium phosphate buffer for the enzyme.

Keywords:

Lactobacilllus, Bifidobacterium, β-galactosidase enzyme activity,

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