Farklı fermente ürünlerden izole edilen laktik asit bakterilerinin alfa-amilaz aktivitesi: Latilactobacillus curvatus Y2-1B amilazının karakterizasyonu

Öz

Bu çalışmada, doksan bir laktik asit bakterisi izolatı farklı fermente ürünlerden elde edilmiş ve α-amilaz üretme yetenekleri nişasta eklenmiş besiyeri kullanılarak kalitatif olarak incelenmiştir. İzole edilen suşlar arasından 75 LAB kalitatif olarak amilaz pozitif kabul edilmiş ve toplam 20 LAB'nin yüksek α-amilaz üretim yeteneğine sahip olduğu belirlenerek kantitatif α-amilaz enzim aktivitesi çalışmaları yapılmıştır. LAB'ler için kantitatif enzim aktivitesi sonuçları 1.283 U/mL- 13.670 U/mL aralığında bulunmuştur. Daha sonra, 3 LAB izolatı (P4-2B, Y2-1B, S1-2B) seçildi ve genotipik yöntemlerle tanımlanmıştır. Tanımlamadan sonra, enzim Latilactobacillus curvatusY2-1B izolatından kısmen saflaştırıldı ve pH, sıcaklık ve katyonların etkisini içeren aktivite analizleri belirlenmiştir. En düşük enzim aktivitesi pH 9'da 0.554 U/ml, en yüksek değer ise pH 6'da 0.694 U/ml olarak bulunmuştur (p<0.05). Öte yandan, saflaştırılan enzim SDS-PAGE'de yaklaşık 40kDa'lık bir bant oluştururken, amy genlerinin varlığı incelenmiş ve onaylanmıştır. Seçilen bu LAB izolatlarının α-amilaz enzim aktivitesinden dolayı endüstriyel biyoteknoloji ve mikrobiyal enzim endüstrisinde faydalı olabileceği değerlendirilmiştir.

Anahtar Kelimeler

• Latilactobacillus curvatus
• amilaz
• spesifik aktivite
• termostabil

Alpha-amylase activity of lactic acid bacteria isolated from different fermented products: Characterization of Latilactobacillus curvatus Y2-1B amylase

Öz

In this study, ninety-one lactic acid bacteria isolates were obtained from different fermented products and their ability to produce α-amylase was qualitatively examined using starch added medium. Among the isolated strains, 75 LABs were qualitatively accepted as amylase positive and a total of 20 LABs were determined to have high α-amylase production ability and quantitative α-amylase enzyme activity studies were performed. Quantitative enzyme activity results for LABs were found in the range of 1.283 U/mL- 13.670 U/mL. Then, 3 LAB isolates (P4-2B, Y2-1B, S1-2B) were selected and identified by genotypic methods. After identification, enzyme was partially purified from Latilactobacillus curvatusY2-1B isolate and activity analyses included the effect on pH, temperature and cations were determined. The lowest enzyme activity was found to be 0.554 U/ml at the pH 9 and 0.694 U/ml at pH 6 as a highest value (p<0.05). On the other hand, the purified enzyme formed a band of approximately 40kDa in SDS-PAGE while the presence of the genes amy were examined and approved. It was thought that these selected LAB isolates may be useful in industrial biotechnology and microbial enzyme industry due to their α-amylase enzyme activity.

Anahtar Kelimeler

• Latilactobacillus curvatus
• amylase
• specific activity
• thermostable

Kaynakça

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