Oxygen transfer enhancement in aqueous/perfluorocarbon fermentation systems: II. Theoretical analysis

Two theoretical approaches were examined to quantify observed experimental oxygen transfer enhancements in aqueous/perfluorocarbon (pfc) fermentation systems. Steady-state macroscopic balance results indicated that enhancements in the presence of reaction were a function of the gas bubble-to-droplet diameter ratio for pfc-in-water dispersions, but primarily a function of only the droplet diameter for water-in-pfc dispersions. The problem of unsteady-state diffusion into a semi-infinite composite medium was employed to predict enhancements in the absence of reaction based on the relative permeabilities of the perfluorocarbon and water phases. Since potential enhancements exceeded actual enhancements in the presence of an oxygen consuming reaction for water-in-pfc dispersions, the use of less expensive, lower oxygenpermeable oil phases is recommended in practice.     

Oxygen transfer in liquid-impelled loop reactors using perfluorocarbon liquids

Oxygen transfer in the liquid-impelled loop reactor is described for a setup in which the perfluorochemical FC40 is aerated externally. Two sizes of reactors are investigated. The mass-transfer coefficient k appears to be lower with a factor of about 0.6 compared to gas liquid systems. the specific exchange area in the present experimental setup is found to be favorable when compared with gas liquid bioreactors at the same superficial dispersed-phase velocities. However, slow coalescence of the dispersed-phase drops in the phase separation section limits the dispersed-phase flow rate seriously. In Case this become crucial from the point of view of oxygen supply, special measures need to be found or alternatives such as combined sparging of air and solvent. (c) 1992 John Wiley & Sons, Inc.     

Oxygen transfer in mechanically agitated aqueous systems containing dispersed hydrocarbon

The effect of dispersed n -dodecane or n -hexadecane on the air-to-aqueous phase overall volumetric oxygen transfer coefficient in a simulated (cell-free) stirred-tank fermentor is described. The oil volume fraction ranged from zero to 0.10; the ionic strength of the aqueous phases was varied from 0 to 0.45. The air-to-aqueous phase coefficients in both oil-free (K_La) and oil-bearing (K_La*) systems were evaluated from unsteady-state experiments using a membrane-covered probe to follow the aqueous phase dissolved oxygen tension. For all systems studied, K_La*/K_La was found to be independent of P/V and v_s for all practical purposes. However, for a particular aqueous phase and at a given P/V and v_s, the ratio K_La*K_La generally differed from unity. Depending on the combination of hydrocarbon type and volume fraction and the aqueous-phase ionic strength employed, the dispersed hydrocarbon may, in some cases, reduce the rate of oxygen transfer and in others enhance it relative to that of the corresponding oil-free gas–liquid dispersion. Enhancement of the air-to-aqueous transfer rate by such negative spreading coefficient hydrocarbons has not been reported previously.     

Oxygen transfer in Streptomyces fermentation broths

The oxygen transfer coefficient has been investigated in S. noursci and S. lavendulae fermentation broths obtained from fermentors of different operating volumes (61., 30001., 20,0001.). Fermentors had K_La_s values ranging from 1.0 to 17.0 min^?1, calculated from sulphite oxidation rates. The dynamic measurement of the volumetric oxygen transfer coefficient. (K_La) has been performed in the different fermenting systems. As the fermentation progressed, especially in the first stages, K_La values have decreased in both fermentations and in each system of fermentors. In order to characterise the whole fermenting system an average K_La_s was calculated from the obtained K_La values. The average K_La grew with increasing K_La_s values and ranged from 0.03 to 3.72 min^?l. Some factors possibly having an influence on the, change of K_La have been studied. The oxygen transfer coefficients of the broths have been measured in falling films and ranged from 0.05 to 0.4 cm min^?1. The flow conditions have been characterized by Reynolds numbers of broths varying between 1.0 and 60.0. The average thickness of the falling films have been measured and plotted against Reynolds number. The Re⁺ which is the breaking-point of the plot increased as the fermentation proceeded. In the region of Re⁺ the values of the oxygen transfer coefficient increased rapidly. An approximate correlation could be established between the Re⁺ and the physical properties of fermentation broth.     

Oxygen mass transfer enhancement via fermentor headspace pressurization.

Bioreactor headspace pressurization represents an excellent means of enhancing oxygen mass transfer to a culture. This method is particularly effective in situations where stirring or vigorous aeration is difficult. Because it in itself introduces no undesirable hydrodynamic force, the proposed method is also attractive for cells susceptible to agitation and sparging. Experiments were first conducted in an ideal fermentor by sparging air into a sulfite solution free from extraneous microbial effects. An increased oxygen mass transfer rate resulting from pressurization led to a superior cell growth rate and a higher maximum cell density in both of the microbial systems studied: a bacterial (Escherichia coli) culture up to 2.72 bar and a fragile algal (Ochromonas malhamensis) culture with pressure programming. Applying pressurization increased the maximum dry cell weight from 1.47 g/L to 1.77 g/L in the E. coli culture and increased the maximum viable cell density from 4 x 10(7) cells/mL to 10(8) cells/mL in the algal culture. An additional advantage is that formation of undesirable products under oxygen limitation, e.g., acetic acid in the E. coli culture, can be suppressed. A significant (over 250%) improvement in the oxygen transfer rate can be achieved with existing fermentors with little modification as they are already designed to withstand reasonable pressure from autoclaving. This method is simple, clean, inexpensive, and easily implemented, and it can be applied alongside other existing methods of oxygen mass transfer enhancement.     

Oxygen transfer kinetics of riboflavin fermentation by Ashbya gossypii in agitated fermentors

Effects of agitation and aeration rates on volumetric oxygen transfer coefficient and oxygen uptake rate of a riboflavin broth containing Ashbya gossypii were investigated in three batch, sparged, and agitated fermentors having the working volumes of 0.42, 0.85, and 2.5 l. The change of oxygen uptake rate with time at 250 rev min⁻¹ stirring and vvm aeration rates was shown. The volumetric oxygen transfer coefficients and maximum oxygen uptake rates obtained have been correlated to mechanical power inputs per unit volume of the fermentation broth and the superficial air velocities.     

Oxygen transfer to mycelial fermentation broths in an airlift fermentor

Oxygen transfer rates and gas holdups were measured in mycelial fermentation broths of Chaetomium cellulolyticum and Neurospora sitophila, each cultured in a 1300-L pilot-plant-scale airlift fermentor. These cultures exhibited highly non-Newtonian flow behavior coupled with a substantial decrease in oxygen transfer rates. The volumetric mass transfer coefficients in these cultures were found to be 65-70% lower than those in water. The data were compared with the available correlations obtained for simulated fermentation broths. In general, the data for C. cellulolyticum are in satisfactory agreement with the correlations for the model media but the data for N. sitophila are higher than that predicted by the correlations. Model media based correlations are found to be applicable to the fermentation processes if the culture medium does not possess a high yield stress.     

Analysis of continuous fermentation processes in aqueous two-phase systems

Simulations of continuous ethanol or acetonobutylic fermentations in aqueous two-phase systems show that at high substrate feed concentrations it is possible to obtain solvent productivities about 25–40% higher than in conventional systems with cell recycle if the biomass bleed rate is kept about one tenth of the value of D.List of Symbols a Volumetric fraction of dextran rich phase - B h^–1 Bleed rate - D h^–1 Dilution rate - P kg m^–3 Product concentration - PD kg m^–3 h^–1 Productivity - S kg m^–3 Substrate - X kg m^–3 Biomass - Partition coefficient     

Oxygen transfer to mixed cultures in tower systems

A modified dynamic method is introduced to determine the oxygen transfer coefficient, K_L a, in aerobic fermentation systems which are not mechanically agitated. The dissolved oxygen concentration is measured continuously following a step down or a step up in aeration rate. The response curve is analyzed to obtain the value of K_La Experiments were carried out at several different air flow rates using mixed culture in concurrent tower fermentors with motionless mixers. The effect of sieve trays and Koch motionless mixers on oxygen transfer was investigated using a 3 in. diameter column. The values of K_L aobtained at the bottom of each column were found to be higher than those obtained at the top. Comparison of the results showed that the values ofK_L a were higher when the Koch mixers were used than when the sieve trays were employed. The oxygen uptake rate by the organisms rX, is also calculated by using the K_L a values obtained. They compare favorably withthe experimentally measured values.     

Intensification of mass transfer in aqueous two-phase systems

A novel technique which intensifies conventional aqueous two-phase extraction by conversion of dispersed phase into colloidal gas aphrons (CGAs) has been developed for extraction of an enzyme. In the present work, amyloglucosidase (1,4-alpha-D-glucan glucohydrolase) was extracted using a polyethylene glycol-sodium sulfate-water system. The lighter phase, i.e., polyethylene glycol (PEG) rich phase, was converted into CGAs which were then dispersed into a salt rich phase. The effect of type of surfactant and its concentration, dispersed phase velocity, phase composition, and type of sparger on the dispersed phase mass transfer coefficient was investigated. The results suggests 9-16 times higher values of mass transfer coefficient compared to spray column. The multiorifice sparger at concentrations of 0.33 g/L of cetyl trimethyl ammonium chloride yielded best results. (c) 1993 John Wiley & Sons, Inc.     

Prehension synergies during nonvertical grasping,I: experimental observations

The mechanical complexities of rotating an object through the gravity field present a formidable challenge to the human central nervous system (CNS). The current study documents the finger force patterns selected by the CNS when performing one-, two-, and four-finger grasping while holding an object statically at various orientations with respect to vertical. Numerous mechanically unnecessary behaviors were observed. These included: nonzero tangential forces for horizontal handle orientations, large internal forces (i.e., those in excess of equilibrium requirements) for all orientations, and safety margins between 50 and 90%. Additionally, none of the investigated measures were constant across orientations or could be represented as a simple trigonometric function of orientation. Nonetheless, all measures varied in systematic (and sometimes symmetric) ways with orientation. The results suggest that the CNS selects force patterns that are based on mechanical principles but also that are not simply related to object orientation. This study is complemented by a second paper that provides an in-depth analysis of the mechanics of nonvertical grasping and accounts for many of the observed results with numerical optimization (see Part II – current issue). Together, the papers demonstrate that the CNS is likely to utilize optimization processes when controlling prehensile actions.Supported in part by NIH grants AR-48563, AG-018751 and NS-35032.     

A mechanistic study of oxygen transfer in aqueous systems

Oxygen transfer coefficients were evaluated for a 14-liter stirred tank fermentor equipped with an oxygen probe, employing elemental copper adsorbed on a weakly basic anion-exchange resin as a solid phase oxygen acceptor. The use of a solid phase oxygen acceptor allowed evaluation of mass transfer resistances associated with the solid phase, and the effect of an oxygen adsorbing solid phase on the overall oxygen transport system, portions of the oxygen transfer process that are neglected by the conventional sulfite oxidation method commonly employed. It was concluded from the data obtained that a transport pathway involving transfer of oxygen to particles present near the air-water interface was a significant oxygen transport pathway for the system studied. Oxygen probe measurements performed on the bulk liquid did not recognize this pathway, suggesting that data taken on biological systems by use of techniques involving oxygen concentration measurements in the bulk liquid may not give the true oxygen absorbing capacity of a system.     

Partition features and renaturation enhancement of chymosin in aqueous two-phase systems

Aqueous two-phase systems of polyethylene glycol (molecular mass 1450, 3350 and 6000)-phosphate and polyethylene-polypropylene oxide (molecular mass 8400)-maltodextrin systems were used in order to study the partition features of recombinant chymosin from inclusion bodies. These systems in the presence of 8M urea were used for the solubilization of inclusion bodies containing recombinant chymosin and for the oxidative renaturation of this protein. Recombinant chymosin showed to be partitioned in favour of the top phase in all studied systems with a partition coefficient between 4 and 6. The recovery of the chymosin biological activity was 32% in the polyethylene-polypropylene oxide, while in the polyethylene glycol-phosphate the recovery was 50-59%. The results indicate that the liquid-liquid extraction would be an adequate tool able to isolate and concentrate chymosin from inclusion bodies with a yield of biological activity higher than that obtained from the standard method (43%).     

Rapid ethanol fermentation of cellulose hydrolysate. I. Batch versus continuous systems

Rapid fermentation of bagasse hydrolysate to ethanol under anaerobic conditions by a strain of Saccharomyces cerevisiae has been studied in batch and continuous cultures at pH 4.0 and 30°C temperature with cell recycle. By using a 23.6 g/liter cell concentration, a concentation of 9.7% (w/v)ethanol was developed in a period of 6 hr. The rate of fermentation was found to increase with supplementation of yeast vitamins in the hydrolysate. In continuous culture employing cell recycle and a 0.127 v/v/m air flow rate, a cell mass concentration of 48.5 g/liter has been achieved. The maximum fermentor productivity of ethanol obtained under these conditions was 32.0 g/liter/hr, which is nearly 7.5 times higher than the normal continuous process without cell recycle and air sparging. The ethanol productivity was found to decrease linearly with ethanol concentration. Conversion of glucose in the hydrolysate to ethanol was achieved with a yield of 95 to 97% of theoretical.     

Delayed fluorescence from Chlorella: I. Low temperature observations

Delayed fluorescence has been observed from Chlorella whole cells at 0.5 ms following flash excitation and at temperatures from 293 °K to 120 °K. Cells which are cooled while pre-illuminated before flashes produce less observed delayed fluorescence than cells cooled without pre-illumination. There exists a small component of delayed fluorescence whose magnitude is independent of pre-illumination effects. The effect of pre-illumination upon delayed fluorescence emission is eliminated by prior freezing of the algae.     

Influence of electrostatic interactions on partitioning in aqueous polyethylene glycol/dextran biphasic systems: Part I

To study the influence of charges on the partition of solutes in aqueous two-phase systems of polyethylene glycol and dextran, partition coefficients of dimethylaminoethyl-dextran, trimethylamino-dextran, and bis (alpha,omega)-amino-poly(ethylene glycol) were determined as a function of pH (range 2 to 12) and ionic strength. These polymers are derivatives of the phase forming components and carry ionizable groups that are charged or uncharged depending on the pH. Unexpectedly, the largest differences in the partition coefficients were found at high pH, where the modified polymers are uncharged. In addition, the partitioning of low-molecular-weight model compounds, ethylenediamine and iminodiacetic acid, as well as poly-L-lysine and poly(allylamine) was analyzed. A consistent pattern was observed in the partition of polyelectrolytes reflecting the influence of charge, but another property of aqueous phase systems unrelated to charge and changing with pH seems to be superimposed. (c) 1995 John Wiley & Sons, Inc.     

Dynamic measurement of the volumetric oxygen transfer coefficient in fermentation systems

A rapid and internally consistent technique has been developed to measure the volumetric oxygen transfer coefficient, k_La, in fermentation systems. The method consists of tracing the dissolved O₂ concentration of the fermentation broth during a short interruption of the aeration. The O₂ concentration trace thus obtained can be analyzed to determine the values of k_La. Additional experiments on prolonged O₂ starvation, carried out to find the limitation of the technique, suggest that O₂ uptake rate will vary if a prolonged (2–10 min.) O₂ starvation occurs.