Katı faz fermantasyonu ile ?-amilaz üretimi ve bazı mantar ekstraktlarının antidiyabetik aktivitelerinin ?-amilazın inhibisyonu ile araştırılması
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Tarih
2021
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Siirt Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada Bitlis Budaklı Kaplıcalarından alınan çamur örneklerinden izole edilen termotolerant VO3 suşu biyolojik materyal olarak kullanıldı. Morfolojik ve biyokimyasal testler ile 16s rRNA gen dizisi analizi sonucu, izolatın Bacillus cinsine ait olduğu ve Bacillus pumilus ile yakından ilişkili olduğu belirlendi. İzolattan katı faz fermantasyon (KFF) yöntemiyle çeşitli tarımsal atıklar (pamuk sapı, mercimek kabuğu, pirinç kabuğu, mısır koçanı ve darı kabuğu) kullanılarak ?-amilaz üretimi amaçlandı. İnkübasyon zamanı, sıcaklık, pH, inokülüm oranı, nem miktarı, karbon ve azot kaynakları gibi farklı parametlerin etkisi araştırıldı. Bacillus pumilus'ten ?-amilaz üretimi için KFF'de uygun substratın pirinç kabuğu olduğu belirlendi. Ayrıca optimum inkübasyon zamanının 48 saat, inkübasyon sıcaklığının 45°C ve başlangıç pH'nın 7.0 olduğu belirlendi. Katı faz fermantasyon ortamına karbon kaynağı olarak nişasta eklendiğinde ve azot kaynağı olarak tripton eklendiğinde enzim üretiminin arttığı tespit edildi. Mantar ekstraktlarının antidiyabetik etkisini belirlemek amacıyla Coprinellus micaceus, Morchella tomentosa ve Helvella queleti'den elde edilen hekzan, metanol ve su ektraktları kullanılarak bu çalışmada üretilen amilaz enziminin inhibisyonu test edildi. En yüksek inhibisyon aktivitesi her üç mantarda da metanol ekstrelerinde görüldü. ?-Amilaz inhibisyon deneylerinde metanol ekstrelerinden C. micaceus, M. tomentosa ve H. queletii'de sırasıyla %37.35, %33.51 ve %28.43 inhibisyon aktivitesi belirlendi.
In this study, thermotolerant VO3 strain isolated from mud samples taken from Bitlis Budaklı Thermal Springs was used as biological material. Morphological and biochemical tests and 16s rRNA gene sequence analysis revealed that the isolate belonged to the genus Bacillus and was closely related to Bacillus pumilus. It was aimed to produce ?-amylase by using various agricultural wastes (cotton straw, lentil husk, rice husk, corn cob and millet husk) from the isolate by solid phase fermentation (KFF) method. The effects of different parameters such as incubation time, temperature, pH, inoculum ratio, moisture content, carbon and nitrogen sources were investigated. It was determined that the suitable substrate in KFF for the production of ?-amylase from Bacillus pumilus was rice husk. In addition, it was determined that the optimum incubation time was 48 hours, the incubation temperature was 45°C and the initial pH was 7.0. It was found that enzyme production increased when starch was added as a carbon source and tryptone was added as a nitrogen source to the solid phase fermentation medium. In order to determine the antidiabetic effect of mushroom extracts, inhibition of amylase enzyme produced in this study was tested by using hexane, methanol and water extracts obtained from Coprinellus micaceus, Morchella tomentosa and Helvella queleti. The highest inhibitory activity was seen in methanol extracts of all three fungi. In ?-Amylase inhibition experiments, 37.35%, 33.51% and 28.43% inhibition activities were determined in C. micaceus, M. tomentosa and H. queletii from methanol extracts, respectively.
In this study, thermotolerant VO3 strain isolated from mud samples taken from Bitlis Budaklı Thermal Springs was used as biological material. Morphological and biochemical tests and 16s rRNA gene sequence analysis revealed that the isolate belonged to the genus Bacillus and was closely related to Bacillus pumilus. It was aimed to produce ?-amylase by using various agricultural wastes (cotton straw, lentil husk, rice husk, corn cob and millet husk) from the isolate by solid phase fermentation (KFF) method. The effects of different parameters such as incubation time, temperature, pH, inoculum ratio, moisture content, carbon and nitrogen sources were investigated. It was determined that the suitable substrate in KFF for the production of ?-amylase from Bacillus pumilus was rice husk. In addition, it was determined that the optimum incubation time was 48 hours, the incubation temperature was 45°C and the initial pH was 7.0. It was found that enzyme production increased when starch was added as a carbon source and tryptone was added as a nitrogen source to the solid phase fermentation medium. In order to determine the antidiabetic effect of mushroom extracts, inhibition of amylase enzyme produced in this study was tested by using hexane, methanol and water extracts obtained from Coprinellus micaceus, Morchella tomentosa and Helvella queleti. The highest inhibitory activity was seen in methanol extracts of all three fungi. In ?-Amylase inhibition experiments, 37.35%, 33.51% and 28.43% inhibition activities were determined in C. micaceus, M. tomentosa and H. queletii from methanol extracts, respectively.
Açıklama
Fen Bilimleri Enstitüsü, Biyoloji Ana Bilim Dalı
Anahtar Kelimeler
Biyoloji, Biology, Biyoteknoloji
