Salda ve Van Göllerinden SGTaz Üreten Alkalifilik Bakterilerin İzolasyonu, Tanılanması ve Enzim Üretme Kapasitelerinin Değerlendirilmesi

Abstract

Bu çalışmanın amacı, CGTase pozitif mikroorganizmaların Salda ve Van göllerinden izolasyonudur. Her iki göl de yüksek seviyede bazik (pH> 9,7) özellikte benzersiz ekstrem ortamlardır. Çalışmada SD5, SG2, SG3 ve V3 olarak kodlanan dört bakteri suşu, seçici Horikoshi katı besiyeri üzerinde yetiştirilerek izole edilmiş ve CGTase üreticileri olarak doğrulanmıştır. Mikroskobik inceleme, dört izolatın Gram pozitif çubuk şeklinde bakteriler olduğunu göstermiştir. Suşlar, 16S rDNA bölgesinin DNA dizilerine dayanarak Bacillus cinsi bakteriler olarak tanılanmıştır. Bunlardan sadece V3 ve SG2 izolatları tür seviyesinde, sırasıyla B. agaradhaerens ve B. patagoniensis olarak tanılanabilmiştir. Suşların büyüme ve CGTase üretim kapasiteleri çalkalamalı inkübatör koşullarında değerlendirilmiştir. Tüm izolatların, 24 saatlik inkübasyon sonrasında hücre dışı ortamda 6-8 U/mL CGTase üretebildiği tespit edilmiştir. Bacillus sp. SG3, Bacillus sp. SD5 ve B. agaradhaerens V3 suşlarının büyüme eğrileri 24-30 saat içerisinde durağan faza ulaşmış ancak 168 saate kadar uzatılmış fermentasyon süresince B. patagoniensis SG2’nin yavaş ancak istikrarlı şekilde çoğaldığı ve CGTase aktivitesinin yükselerek devam ettiği gözlemlenmiştir. Hücredışı CGTase salgılama kapasitesi olduğu doğrulanan bu dört bakteri suşu gelecekte CGTase'in endüstriyel ölçekte üretilebilmesi için yürütülecek araştırmalarda potansiyel gen kaynakları olarak değerlendirilebilecektir.

Keywords

• Bacillus sp.
• bakteriyel tanılama
• siklodekstrin glikosiltransferaz
• alkalifilik

Isolation, Identification and Evaluation of Enzyme Production Capacity of CGTase Producing Bacteria from Lakes Salda and Van/Turkey

Abstract

The aim of this study was isolation of CGTase positive microorganisms from lakes Salda and Van, both being extreme environments unique in character with high alkalinity pH>9.7. Four bacterial strains designated as SD5, SG2, SG3 and V3 were isolated by cultivation on selective Horikoshi agar and confirmed as CGTase producers. Microscopic inspection showed the four isolates to be Gram positive rods. Strains were identified as Bacillus based on DNA sequences of 16S rDNA region. Only the V3 and SG2 isolates could be resolved at species level identified as B. agaradhaerens and B. patagoniensis, respectively. Growth and CGTase production capacity of the strains were evaluated under shake flask conditions. All isolates achieved extracellular CGTase levels of 6-8 U/mL within 24 h of incubation. Growth curves of Bacillus sp. SG3, Bacillus sp. SD5, B. agaradhaerens V3 showed the bacteria to reach a plateau phase within 24-30 h however, extended fermentation up to 168 h showed B. patagoniensis SG2 to continue the growth cycle until the end of this period with higher CGTase activity observed. These four strains isolated which have thus been confirmed as CGTase positive bacteria can potentially be exploited as genetic sources for the synthesis of CGTase in industrial scale.

Keywords

• Bacillus sp.
• bacterial identification
• cyclodextrin glycosyltransferase
• alkaliphilic

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