Aktinobakteri İzolatlarının Transglutaminaz, Levansukraz ve Beta Galaktozidaz Üretim Yetenekleri

Year 2022, Volume: 20 Issue: 1, 30 - 39, 03.04.2022

Elif Gülşen Karabacak , Ali Osman Adıgüzel Hayrettin Saygın Ahmet Hilmi Çon

https://doi.org/10.24323/akademik-gida.1097824

Abstract

Aktinobakteriler ekstrem şartlarda gelişme, büyük miktarlarda enzim üretme potansiyeli, biyokimyasal çeşitlik ve genetik manipülasyonlara uygunluk özellikleriyle alternatif enzim kaynakları arasında önemli bir konumdadır. Çalışmada, endüstriyel alanda kullanımı fazla olan transglutaminaz, β-galaktozidaz ve levansukraz enzimleri için uygun bir üretici Aktinobakteri cinsi mikroorganizmanın seçilmesi hedeflenmiştir. Bu amaçla ekstrem koşullara sahip habitatlardan farklı araştırmacılar tarafından izole edilmiş 46 aktinobakteri izolatının hedeflenen enzimleri üretim yetenekleri araştırılmıştır. Aktinobakteri izolatlarının ilgili enzimler açısından üretici olup olmadıkları, önce veri tabanlarında kayıtlı olan genom dizilerinin “Rapid Annotation using Subsystem Technology Version 2.0” kullanılarak taranmış, devamında ilgili gene sahip olanların transglutaminaz için Hidroksimat Yöntemi (Kağıt Disk Yöntemiyle), β-galaktozidaz için ONPG yöntemi, levansukraz için ise mukoid yapı oluşturma fenotipinin belirlenmesi şeklinde enzim üretme yetenekleri belirlenmiştir. Biyoinformatik taramada tüm izolatların “transglutaminaz benzeri enzim” kodlayan gen bölgesi içerdiği, kalitatif tarama sonucunda farklı türe sahip ve besiyerinde daha hızlı gelişim gösteren 9 adet bakteri izolatının potansiyel olduğu belirlenmiştir. Levansukraz enzim genine ise sadece Micromonospora sp. KC721 ve Micromonospora sp. KC213 izolatlarının sahip olduğu ancak hiçbir izolat ne katı ne de sıvı besiyerinde aktivite göstermemiştir. ß-Galaktozidaz enzim üretim geni varlığı 38 izolatta saptanmıştır. Enzim üretim genine sahip izolatlara uygulanan kalitatif test sonucunda, daha yoğun renk oluşturan, farklı türe sahip olan ve besiyerinde diğerlerine göre hızlı gelişim gösteren 17 izolat potansiyel β-galaktozidaz üreticisi olarak seçilmiş ve farklı biyoteknolojik uygulamalar için endüstriyel ölçekli enzim üretiminde kullanım potansiyeline sahip aktinobakter izolatları olarak belirlenmiştir.

Keywords

Enzim üretim yeteneği, Transglutaminaz, Levansukraz, ß-Galaktozidaz, Aktinobakteri

Production Capabilities of Transglutaminase, Levansucrase and Beta Galactosidase of Actinobacteria Isolates

Abstract

Actinobacteria occupy an important position among alternative enzyme sources with their ability to growth in extreme conditions, their potential to produce large amounts of enzymes, biochemical diversity and compatibility with genetic manipulations. In this study, the selection of suitable producer microorganisms for transglutaminase, ß-galactosidase and levansucrase enzymes was aimed, and the enzyme production capabilities of 46 actinobacterial isolates were determined. The ability of these actinobacterial isolates to produce related enzymes was determined by scanning the genome sequences deposited in “GenBank using Rapid Annotation using Subsystem Technology Version 2.0”. Subsequently, the enzyme production capabilities of these isolates with relevant gene were determined by Hydroximate Method for transglutaminase (with the Paper Disc Method), the ONPG method for ß- galactosidase and the mucoid structure formation phenotype for levansucrase. By bioinformatics scanning, it was determined that all isolates contained a “transglutaminase-like enzyme” gene region and by qualitative screening, 9 isolates with different species and that can grow rapidly in medium were identified as potential isolates. A levansucrase-production gene was determined in only 2 isolates (Micromonospora sp. KC 721 and Micromonospora sp. KC213) but none of these showed activity in agar and broth medium. The presence of ß-galactosidase enzyme production gene was detected in 38 isolates. As a result of the qualitative test, 17 isolates with more intense color, different species and that can grow rapidly in medium were selected as potential ß-galactosidase-producers and determined as actinobacterial isolates with a potential to be used in industrial scale enzyme production for different biotechnological applications.

Keywords

Enzyme production capability, Transglutaminase, Levansucrase, ß-Galactosidase, Actinobacteria

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