Yerli Alfa-Amilaz Enzimi Üretimi İçin Yerli Fungus İzolasyonu, Makroskobik, Mikroskobik Tanımlaması, DNA Dizi Analizi ve Biyoinformatik Değerlendirmeler

Abstract

Bu çalışmada ekmek yapımında ticari önemi olan alfa-amilaz enzimi üreten termofilik ve mezofilik yerli fungusların izolasyonu, makroskobik, mikroskobik ve moleküler tür teşhisleri gerçekleştirilmiştir. Mikroorganizma Kültür Koleksiyon Merkezlerine ve Mikrobiyal Gen Bankalarına genetik materyal oluşturulması ve yerli izolatlardan patente konu yerli enzim üretimi amaçlanmıştır. Afyon, Eskişehir, Uşak, Ankara termal alanlardan 23 termotolerant fungus izole edilmiştir. İzolatların makroskobik ve mikroskobik incelemeleri sonucunda yüksek amilaz enzimi üreten Aspergillus niger, Aspergillus terreus ve Trichoderma atroviride türüne ait 6 izolatın tür teşhisi yapılmıştır. İzolatların fungus spesifik primerleri kullanılarak 18S rDNA ve ITS bölgeleri PCR amplifikasyonu, moleküler tanımlaması yapılmış ve Web tabanlı BLAST analizleri ile karşılaştırılmıştır. Moleküler karakterizasyon çalışması sonucunda klasik tanımlama ile moleküler tanımlamanın birbirini desteklediği görülmüştür.

Keywords

• Fungus
• izolasyon
• Internal Transcribed Spacer (ITS)
• amilaz enzimi

Isolation, Macroscopic, Microscopic Identification, DNA Sequencing and Bioinformatic Assessments of Native Fungi for the Production of Native Alpha-Amylase Enzyme

Abstract

In this study, isolation of thermophilic and mesophilic native fungi producing alpha-amylase enzyme which is commercially important in bread making, macroscopic, microscopic and molecular identification were performed. Microorganism Culture Collection Centers and Microbial GenBanks to create genetic material and local isolates to produce patents of the native enzyme was aimed. 23 thermotolerant fungi were isolated from the thermal areas of Afyon, Eskişehir, Uşak and Ankara. As a result of macroscopic and microscopic investigations of isolates, 6 isolates of Aspergillus niger, Aspergillus terreus and Trichoderma atroviride species which produce high amylase enzyme species were determined. 18S rDNA and ITS regions of the isolates were compared with Web-based BLAST analysis by PCR amplification and molecular identification. As a result of molecular characterization study, it was observed that the classical identification and the molecular identification supported each other.

Keywords

• Fungi
• isolation
• Internal Transcribed Spacer (ITS)
• amylase enzyme

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