Transferência de oxigênio e condições de cisalhamento em biorreator convencional com impelidores orelha de elefante
Bustamante, Maritza Catalina Condori
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Due to owing to the nature of culture involving aerobic filamentous microorganisms, conventional aerated stirred-tank bioreactors using the Rushton turbine impeller are widely employed in order to achieve good mixing and oxygen mass transfer. However, this impeller presents disadvantages such as high power consumption, and generates higher shear in the medium, which can cause irreversible morphological changes in microorganisms, damaging the culture performance. In literature, impellers called "Elephant Ear" (EE), presented in down-flow (down-pumping) (EEDP) and up-flow (up-pumping) (EEUP) configurations, are indicated as suitable for cultivation of shear sensitive cells. However, this characteristic has never been demonstrated quantitatively. In the present study, the oxygen transfer and hydrodynamics parameters were evaluated and compared in a conventional aerated stirredtank bioreactor equipped with impellers as the classical impeller Rushton turbine (RT) and with EE in EEDP and EEUP modes. Firstly, prediction equations were obtained for quantifying the average shear rate (Ýav) in the different systems, as function of the operating conditions (600≤N≤1000 rpm and 0.40≤Ýavar≤1.2 vvm) and rheological properties of the pseudoplastic fluids (K and n), based on a methodology that uses the volumetric oxygen transfer coefficient as characteristic parameter. The values of Ýav obtained from the correlations for the RT impeller were within a range of values estimated from correlations of the literature, showing that the methodology utilized is valid. Based on correlations of Ýav and on results of power consumption, the energy dissipation and Kolmogorov micro-scale (micro eddy sizes) could be estimated. For non-Newtonian fluids, the RT and EEUP impellers generated micro-eddies with smaller sizes in the range from 45.0-127.0 and 80.6-123.7 μm, respectively, showing that the impellers RT and EEUP are more shear than EEDP impeller. Using the prediction equations of Ýav the effect of shear conditions in cultures of Streptomyces clavuligerus were evaluated in conventional bioreactor equipped with RT, EEDP and EEUP impellers at 800 rpm, 0.50 vvm and 30C. Profiles of the volumetric oxygen transfer coefficient and shear rate were obtained during the cultivation. The morphological changes resulted from shear conditions imposed for the different impellers were analyzed using image analysis technique. The RT and EEDP impellers imposed the highest oxygen transfer and smaller shear to the broth, respectively. The flow pattern generated by EE impellers was crucial at the beginning of the cultivation, and is highly beneficial when EEUP impeller were used, since it presented a controlled shear throughout the cultivation. The best cultivation condition for the clavulanic acid production was obtained using the EEUP impeller, which resulted in a maximum concentration of 453 mg/L and a consistency index K= 2.5 Pa.s0.27. The results suggested that the isolated hyphae are directly related to the K of the broth and clavulanic acid production. According to the rapid fragmentation of clumps and emergence of isolated hyphae, it was concluded that the RT impeller presents shear rates higher than the EEs impellers, since the size of branched and non-branched hyphae were smaller and in greater quantities. This fact shows an advantage in the EEUP impeller use in shear sensitive microorganism cultures that requiring high levels of oxygen, without affecting the process performance. This advantage is associated with the flow pattern generated for the EEUP impeller.