Vitreoscilla hemoglobin eksprasyonu yapan rekombinant Erwinia herbicola'da ikincil metabolit akış dağılımı
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Dosyalar
Tarih
2012
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İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada, Erwinia herbicola ve onun Vitreoscilla hemoglobin geni (vgb) klonlanmış suşu (vgb+) ile vgb içermeyen (vgb-) rekombinant suşları kullanılmıştır. Vitreoscilla hemoglobinin (VHb) E. herbicola'da indol-3-asetik asit, ß-karoten, likopen, C vitamini, fenazin, meropenem, ATP ve NAD üretimi üzerindeki etkisi araştırılmıştır. E. herbicola'da bu şekilde endojen rekombinant bir oksijen alım sisteminin bu metabolitlerin üretimi üzerindeki etkisi ilk defa çalışılmıştır. Diğer iki suşla karşılaştırıldığında, vgb+ rekombinant suşunun düşük konsantrasyonda L-triptofan eklenen zengin kültür ortamının ileri kültür fazı ile yüksek glukozlu M9 ortamında önemli derecede yüksek indol-3-asetik asit seviyesine sahip olduğu belirlendi. Yine bu suşun diğer suşlara göre zengin besiyerinde daha yüksek ß-karoten ve likopen seviyesine sahip olduğu gözlendi. VHb'nin C vitamini üretimini hem glukozlu M9 ortamı ve hem de zengin besiyerinde pozitif yönde etkilediği saptanırken, fenazin üretiminin fruktoz ortamında ve meropenem üretiminin ise sükroz ortamında VHb varlığından pozitif yönde etkilendiğ belirlendi. Ayrıca, rekombinant suşların konakçıdan önemli derecede yüksek oksijen alımına sahip oldukları belirlendi. Zengin besiyerinde vgb+ suşunun hem ATP hem de NAD düzeyleri konakçı ve vgb- rekombinantından daha yüksek bulundu.
In this study Erwinia herbicola and its recombinants, one carrying the Vitreoscilla hemoglobin gene (vgb+) and the other missing it (vgb-), were used in a comparative fashion with respect to production of selected secondary metabolites. In this regard, this study is the first for investigation of the effect of such an endogenous recombinant oxygen-uptake system on microbial production of secondary metabolites as indol-3-asetik asit, ß-caroten, lycopene, vitamin C, phenazine, meropenem whose production are tightly regulated by oxygen. Further, the energetic state of bacteria were evaluated through the level of ATP and NAD. Compared to other two strains, the level of indol-3-asetik asit in the low L-tryptophan rich medium late phase vgb+ cultures was considerably high. Similarly, the level of this hormone in high glucose minimal medium cultures of vgb+ was higher than the host and vgb- strains. The rich medium cultures of vgb+ recombinant strain had also considerably higher levels of ß-carotene and lycopene. The vgb+ strain was also determined to have a higher level of vitamin C both in rich and glucose supplemented minimal media cultures. The presence of vgb was also determined to positively regulate the production of two antibiotics, phenazine and meropenem, under different carbon supply. The highest phenazine and meropenem levels were achieved with fructose and sucrose, respectively. Also, the recombinants had considerably higher oxygen uptake rates than the host strain. The levels of both ATP and NAD in rich cultures of vgb+ recombinant were higher than the host and vgb- recombinant.
In this study Erwinia herbicola and its recombinants, one carrying the Vitreoscilla hemoglobin gene (vgb+) and the other missing it (vgb-), were used in a comparative fashion with respect to production of selected secondary metabolites. In this regard, this study is the first for investigation of the effect of such an endogenous recombinant oxygen-uptake system on microbial production of secondary metabolites as indol-3-asetik asit, ß-caroten, lycopene, vitamin C, phenazine, meropenem whose production are tightly regulated by oxygen. Further, the energetic state of bacteria were evaluated through the level of ATP and NAD. Compared to other two strains, the level of indol-3-asetik asit in the low L-tryptophan rich medium late phase vgb+ cultures was considerably high. Similarly, the level of this hormone in high glucose minimal medium cultures of vgb+ was higher than the host and vgb- strains. The rich medium cultures of vgb+ recombinant strain had also considerably higher levels of ß-carotene and lycopene. The vgb+ strain was also determined to have a higher level of vitamin C both in rich and glucose supplemented minimal media cultures. The presence of vgb was also determined to positively regulate the production of two antibiotics, phenazine and meropenem, under different carbon supply. The highest phenazine and meropenem levels were achieved with fructose and sucrose, respectively. Also, the recombinants had considerably higher oxygen uptake rates than the host strain. The levels of both ATP and NAD in rich cultures of vgb+ recombinant were higher than the host and vgb- recombinant.
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Kurt, A. G. (2012). Vitreoscilla hemoglobin eksprasyonu yapan rekombinant Erwinia herbicola'da ikincil metabolit akış dağılımı. İnönü Üniversitesi Fen Bilimleri Enstitüsü. 1-159 ss.
