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The effect of nitrite on enhancement of alpha-amylase synthesis afforded by bacterial hemeglobin in genetically enginered E. Coli

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  1. Tez No: 400514
  2. Yazar: SÜLEYMAN AYDIN
  3. Danışmanlar: PROF. DR. BENJAMIN C. STARK, PROF. DR. DALE WEBSTER
  4. Tez Türü: Doktora
  5. Konular: Hematoloji, Klinik Bakteriyoloji ve Enfeksiyon Hastalıkları, Hematology, Clinical Microbiology and Infectious Diseases
  6. Anahtar Kelimeler: Belirtilmemiş.
  7. Yıl: 1999
  8. Dil: İngilizce
  9. Üniversite: Illinois Institute of Technology
  10. Enstitü: Yurtdışı Enstitü
  11. Ana Bilim Dalı: Belirtilmemiş.
  12. Bilim Dalı: Belirtilmemiş.
  13. Sayfa Sayısı: 104

Özet

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Özet (Çeviri)

Geneticaily engineering heterelogus bacterial hosts with the Vitreoscilla hemoglobin gene (vgb) so that they produce Vitreoscilla hemoglobin (VHb) has been shown to provide advantages in both grovvth and product production, particularly under hypoxic condİtions. An example is the matched set of Ecoli strains MK79 and MK57 wbich carry plasmids pMK79 and pMK57, respectively. Plasmid pMK57 carries the Bacillus stearothermophilus alpha-amylase gene in vector pUC8; pMK79 is identical except it also carries vgb. MK79 outgrows MK57 and produces signifıcanüy more alpha-amylase than MK57, especially under hypoxic condİtions. The objective of this work was to specifically inactivate VHb with nitrite, which oxidizes it to the met form, and determine the effect of this inactivation on plasmid stability, biomass'production, oxygen consumption and alpha-amylase production of strain MK79 in comparison to strain MK57. Accumulation of active VHb was inhibited and VHb oxidİzed to the inactive ferric form by nitrite in the range of 0.5 to 40 mM, concentration which had little effect on the cytochromes of the respiratory chain. This decrease in active VHb in strain MK79 was correlated with decreases in alpha-amylase production, oxygen consumption and celi mass that were more severe than those suffered by its counterpart, MK57 (lacking VHb). A“crossover”point occurred at 5 mM nitrite at which the advantages of MK79 över MK57 due to vgb/VHb disappeared, and then become disadvantages at 20 mM nitrite and above. Unexpectedly, for growth in nitrite at 5 mM or less, but above zero nitrite, largeincreases in alpha-amylase activity occurred for both strains compared to growth in the absence of nitrite. it is concluded that vgb, and not any other Vitreoscilla DNA sequences on plasmid pMK79 iş the source of the advantages in growth and recombinant protein (alpha-amylase) production in this system, and by extension in other systems using vgö/VHb.

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