Proteaz üreticisi mikroorganizmaların 16s rrna ile moleküler tanımlanması ve bu mikroorganizmaların proteaz enzimleri üzerine çalışmalar
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Dosyalar
Tarih
2017
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Kırıkkale Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada, proteince zengin olan bir bölgeden alınan toprak örneğindeki proteaz enzimi üreten mikroorganizmalar saflaştırılmış ve bu mikroorganizmaların üretmiş olduğu proteaz enzimi ile çalışmalar yapılmıştır. İzole edilen mikroorganizmaların en uygun üreme şartları belirlenlenmiş ve biyokimyasal testleri yapılmıştır. 16S rRNA analizi yapılan mikroorganizmaların 5 tanesi Bacillus cinsine ait, 1 mikroorganizma ise Aeromonas salmonicida olarak tanımlanmıştır. En uygun enzim üretim şartlarında 82,156 U/mL ile en yüksek enzim aktivitesini gösteren Aeromonas salmonicida'dır. Enzim çalışmalarına enzim aktivitesi 60,5 U/mL olan en yüksek 2. aktiviteli ORSK-4 (Bacillus cereus) ile devam edilmiştir. Elde edilen proteaz enziminin en uygun çalışma sıcaklığı 27 ile 60 °C arasında, en uygun çalışma pH'sı ise pH: 9.0 olarak belirlenmiştir. Mn2+ iyonunun enzim aktivatörü olduğu, benzen ve H2O2'nin enzim aktivitesini arttırdığı belirlenmiştir. Enzim EDTA ile % 72,28 oranında inhibe olmuştur. Enzimin moleküler ağırlığı SDS PAGE analizinde 20 ve 30 kDa olarak bulunmuştur.
In this study, the microorganisms of producing protease enzymes were purified from a soil rich in protein and the protease enzyme produced by these microorganisms has been studied. The optimum growth conditions of isolated microorganisms were determined and biochemical tests were performed. 5 isolates were identified as Bacillus species and 1 isolate was identified as Aeromonas salmonicida by using 16S RNA gene analyses. Aeromonas salmonicida is shows the highest enzyme activity with 82,156 U/mL under optimal enzyme production conditions. Enzyme studies continued with ORSK-4 (Bacillus cereus) that has the second highest enzyme activity as 60.5 U/mL. The optimum temperature of the obtained protease enzyme was determined to be between 27 and 60 ° C and the optimum pH was 9.0. It is determined that Mn 2 + ion is the enzyme activator and that the enzyme activity of benzene and H2O2 is increased. The enzyme was inhibited 72.28 % with EDTA. The molecular weight of the enzyme has found 20 and 30 kDa by SDS PAGE analysis.
In this study, the microorganisms of producing protease enzymes were purified from a soil rich in protein and the protease enzyme produced by these microorganisms has been studied. The optimum growth conditions of isolated microorganisms were determined and biochemical tests were performed. 5 isolates were identified as Bacillus species and 1 isolate was identified as Aeromonas salmonicida by using 16S RNA gene analyses. Aeromonas salmonicida is shows the highest enzyme activity with 82,156 U/mL under optimal enzyme production conditions. Enzyme studies continued with ORSK-4 (Bacillus cereus) that has the second highest enzyme activity as 60.5 U/mL. The optimum temperature of the obtained protease enzyme was determined to be between 27 and 60 ° C and the optimum pH was 9.0. It is determined that Mn 2 + ion is the enzyme activator and that the enzyme activity of benzene and H2O2 is increased. The enzyme was inhibited 72.28 % with EDTA. The molecular weight of the enzyme has found 20 and 30 kDa by SDS PAGE analysis.
Açıklama
Anahtar Kelimeler
Biyoloji, Biology
