Mutant Bacillus megaterium bakterisinin biyomineralizasyon kapasitesinin araştırılması
Yükleniyor...
Dosyalar
Tarih
2023
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Bursa Teknik Üniversitesi, Lisansüstü Eğitim Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Yapılarda oluşan bozulmaların iyileştirilmesi için kullanılan geleneksel yöntemlerin çoğunlukla yetersiz olup zaman içerisinde farklı bozunma sorunlarına yol açtığı bilinmektedir. Bu nedenle son yıllarda kullanılan en yenilikçi yöntemlerden biri biyominerilizasyondur. Mikrobiyal kaynaklı kalsit çökeltme (MICP) işlemi, yapının dayanıklılığını artıran ve orijinal mikroyapıya göre onarıp iyileştirmesini sağlayan bir biyomineralizasyon yöntemidir. Ürolitik bakteriler tarafından üreaz enzimi ile gerçekleştirilen üre hidrolizi, daha yüksek konsantrasyonda çökeltilmiş karbonat elde etmede kalsitleşme için önemli bir faktördür. Bu araştırma kapsamında ürolitik ve patojenik olmayan Bacillus megaterium türü bakteri ile çalışmalar gerçekleştirilmiştir. B. megaterium, 10x35x20 cm boyutlarında hazırlanmış kutu içinde 8-watt ultraviyole floresan lamba kullanılarak sırasıyla 8 sn, 11 sn, 13 sn, 15 sn, 17 sn ve 21 sn boyunca UV ışımaya maruz bırakılmıştır. Oluşan mutant bakterilerin üreaz enzim aktivasyonları ile orijinal bakterinin üreaz enzim aktivitesini karşılaştırmak için Christensen Urea Agar Base ve Urease Activity Assay Kit (BioVision, K378-100) kullanılmıştır. Christensen Urea Agar Base kullanılarak yapılan çalışmada, mutantlar ve kontrol olarak kullanılan orijinal tür üre agara ekilerek 24 sa., 48 sa. ve 72 sa. boyunca inkübe edilmiştir. İnkübasyon süresi sonlarında oluşan magenta renginin değişimleri fotoğraflanarak, renk yoğunlukları ölçülmüştür. Oluşan renk yoğunluklarının mutant bakterilerde kontrole göre daha fazla olduğu ve yoğunluğun UV ışımaya maruz kalma süresiyle doğru orantılı olarak arttığı belirlenmiştir. Belirlenen sonuçların sağlamasını yapmak amacıyla üreaz enzimi aktivitesi ölçen kit kullanılarak, B. megaterium bakteri suşu ve 21 sn UV ışıma ile oluşan mutant bakterinin enzim aktiviteleri test edilmiştir. Üreaz enzimi aktivitesine bağlı amonyak oluşumu konsantrasyonunun B. megaterium için 1.01 nmol, mutant bakteri için 3.32 nmol olduğu ölçülmüştür. Oluşan amonyak miktarları incelendiğinde konsantrasyon ortalamaları arasında % 95 oranla (P<0,05) anlamlı bir farkın olduğu ve mutantın istatiksel olarak daha fazla amonyak ürettiği görülmüştür. Çalışma sonunda 21 sn UV ışımaya maruz kalan bakterinin yüksek pH şartlarına adaptasyonun arttığı ve üreaz enzim aktivitesinin yükseldiği anlaşılmıştır. Buna bağlı olarakta mutant bakterinin orijinal bakteriye oranla kalsifikasyon potansiyelinin ve biyomineralizasyon kapasitesi arttığı belirlenmiştir.
It is known that the traditional methods used to improve the deterioration of structures are mostly inadequate and cause different deterioration problems over time. Therefore, one of the most innovative methods used in recent years is biomineralization. Microbial calcite precipitation (MICP) is a biomineralization method that increases the durability of the structure and allows it to repair and improve according to the original microstructure. Urea hydrolysis by urease enzyme by urolytic bacteria is an important factor for calcification to obtain a higher concentration of precipitated carbonate. Within the scope of this research, studies were carried out with urolytic and non-pathogenic bacteria Bacillus megaterium. The bacteria were exposed to UV irradiation for 8 seconds, 11 seconds, 13 seconds, 15 seconds, 17 seconds, and 21 seconds, respectively, using an 8-watt ultraviolet fluorescent lamp in a 10x35x20 cm box. Christensen Urea Agar Base and Urease Activity Assay Kit (BioVision, K378-100) were used to compare the urease enzyme activations of the mutant bacteria with the urease enzyme activity of the original bacteria. In the study using Christensen Urea Agar Base, mutants and the original strain used as control were cultivated on urea agar for 24 h, 48 h. and 72 h. incubated throughout. At the end of the incubation period, the changes in the magenta color were photographed and the color intensities were measured. It was determined that the color intensities formed were higher in mutant bacteria than in the control and the intensity increased in direct proportion to the exposure time to UV radiation. In order to ensure the determined results, the enzyme activities of the B. megaterium bacterial strain and the mutant bacteria formed by 21 sec UV irradiation were tested by using the kit that measures the urease enzyme activity. The concentration of ammonia formation due to urease enzyme activity was measured as 1.01 nmol for B. megaterium and 3.32 nmol for mutant bacteria. When the amount of ammonia formed was examined, it was seen that there was a significant difference of 95% (P<0.05) between the concentration averages and the mutant produced statistically more ammonia. At the end of the study, it was understood that the adaptation to high pH conditions of the bacteria exposed to UV radiation for 21 seconds increased and the urease enzyme activity increased. Accordingly, it was determined that the calcification potential and biomineralization capacity of the mutant bacteria increased compared to the original bacteria.
It is known that the traditional methods used to improve the deterioration of structures are mostly inadequate and cause different deterioration problems over time. Therefore, one of the most innovative methods used in recent years is biomineralization. Microbial calcite precipitation (MICP) is a biomineralization method that increases the durability of the structure and allows it to repair and improve according to the original microstructure. Urea hydrolysis by urease enzyme by urolytic bacteria is an important factor for calcification to obtain a higher concentration of precipitated carbonate. Within the scope of this research, studies were carried out with urolytic and non-pathogenic bacteria Bacillus megaterium. The bacteria were exposed to UV irradiation for 8 seconds, 11 seconds, 13 seconds, 15 seconds, 17 seconds, and 21 seconds, respectively, using an 8-watt ultraviolet fluorescent lamp in a 10x35x20 cm box. Christensen Urea Agar Base and Urease Activity Assay Kit (BioVision, K378-100) were used to compare the urease enzyme activations of the mutant bacteria with the urease enzyme activity of the original bacteria. In the study using Christensen Urea Agar Base, mutants and the original strain used as control were cultivated on urea agar for 24 h, 48 h. and 72 h. incubated throughout. At the end of the incubation period, the changes in the magenta color were photographed and the color intensities were measured. It was determined that the color intensities formed were higher in mutant bacteria than in the control and the intensity increased in direct proportion to the exposure time to UV radiation. In order to ensure the determined results, the enzyme activities of the B. megaterium bacterial strain and the mutant bacteria formed by 21 sec UV irradiation were tested by using the kit that measures the urease enzyme activity. The concentration of ammonia formation due to urease enzyme activity was measured as 1.01 nmol for B. megaterium and 3.32 nmol for mutant bacteria. When the amount of ammonia formed was examined, it was seen that there was a significant difference of 95% (P<0.05) between the concentration averages and the mutant produced statistically more ammonia. At the end of the study, it was understood that the adaptation to high pH conditions of the bacteria exposed to UV radiation for 21 seconds increased and the urease enzyme activity increased. Accordingly, it was determined that the calcification potential and biomineralization capacity of the mutant bacteria increased compared to the original bacteria.
Açıklama
Anahtar Kelimeler
Bacillus megaterium, Biyo-mineralizasyon, Mutasyon, UV radyasyon, MICP, Kalsifikasyon, Bio-mineralization, Mutation, UV radiation, Calcification
