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.     

Extractive recovery of products from fermentation broths

Considerations of partition coefficients, selectivity, biocompatibility, and waste generation are important in selection of appropriate solvents to be used for extractive recovery of products from fermentation broths. Several selection criteria can be used based upon the nature of different species present in the broth. These criteria, along with examples of specific case studies, were presented. These serve not only in screening of useful solvents, but also in pointing to the specific modes of operation of recovery-coupled bioprocesses.     

Oxygen-stabilized enzyme electrode for d-glucose analysis in fermentation broths

A new principle for the construction of oxygen-dependent enzyme electrodes is presented. The enzyme electrode is based on a galvanic oxygen electrode which is furnished with an electrolysis anode covered by immobilized enzyme and placed close to the oxygen-sensing surface. An ordinary oxygen electrode is used as a reference electrode. The enzymatic consumption of oxygen in the enzyme electrode generates a potential difference between the electrodes which is utilized to control electrolytic generation of oxygen from water in such a way that zero differential potential is maintained. Thus, the enzyme electrode operates under ambient oxygen tension and does not suffer from oxygen limitation. The electrolytic current in this system gives a measure of the concentration of substrate surrounding the enzyme electrode. The electrode has been applied for continuous d-glucose analysis in situ during batch cultures of Candida utilis.     

Spectrophotometric method for determining gibberellic acid in fermentation broths

A novel method for the quantitative determination of gibberellic acid in fermentation broths has been developed. It is based on the kinetic of the reaction of conversion of gibberellic acid to gibberellenic acid. The method is simple, reliable, faster than most of methods known, and free of the interferences which commonly affect spectrophotometric methods currently in use. Its threshold sensitivity is 0.1 g and its accuracy is greater than 97% for concentrations of gibberellic acid ranging from 0.1 to 1 g l(-1).     

Measurement of penicillin acylase and cephalosporin acylase in fermentation broths

Summary The p-dimethylamino-benzaldehyde method of Bomstein and Evans, the ninhydrin method of Marrelli and the new p-dimethyl-amino-benzaldehyde method of Kornfeld were evaluated for analysis of penicillin-V acylase and cephalosporin-V acylase in fermentation broths. The ninhydrin method was the method of choice for cephalosporin-V acylase, whereas the Kornfeld method had certain advantages with respect to penicillin-V acylase.     

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.     

Procedure for isolation of Thienamycin from fermentation broths

An improved process for the isolation of thienamycin, produced by the actinomycete Streptomyces cattleya, has been developed. The isolation procedure consists of three chromatographic steps, volume reduction by reverse osmosis between the steps, and freezedrying for obtaining the final product. The chromatographic steps are as follows: (1) ion exchange chromatography on Dowex 1 × 2 resin in the bicarbonate cycle, (2) gel chromatography on Dowex 1 × 2 resin in the chloride cycle, (3) reverse phase chromatography on XAD-2 resin. This procedure is useful for processing large volumes of fermentation broth.     

Enhancement of oxygen transfer in highly viscous non-newtonian fermentation broths

The influence of short draft tubes covered by perforated plates on gas-liquid mass transfer was examined in external-loop airlift bioreactors. The volumetric mass transfer coefficients in a model external-loop airlift bioreactor were measured with water and non-Newtonian media. It was found that introduction of draft tubes covered with perforated plates in the riser significantly improved the mass transfer rate, particularly in higher viscous non-Newtonian fermentation media. The enhancement of mass transfer rate might be due mainly to an increase in bubble coalescence and redispersion. (c) 1994 John Wiley & Sons, Inc.     

Recovery of ethanol from fermentation broths using selective sorption-desorption

Industrialized nations face a critical problem in replacing the sources of liquid fuels that traditionally have been supplied by petroleum. One solution that has gained increasing support in this country is the use of ethanol produced by fermentation of renewable biomass as an extender in, or supplement to, gasoline for transportation fuel. Distillation, the present method of separating ethanol from the fermentation broth, is an energy-intensive one and frequently uses more energy than is available from the ethanol recovered. There are many investigations under way to find alternative, less energy-intensive techniques for the ethanol-water separation. The separations method described in this article involves the use of solid materials to preferentially remove ethanol from fermentation broths. Subsequent stripping of the ethanol from the sorbent with a dry gas reduces dramatically the energy required for the separation. Three solid sorbents have been investigated experimentally. Their sorption/desorption characteristics are described, and their incorporation in an ethanol recovery process is evaluated. Three sorbents were investigated: two commercially available divinylbenzene crosslinked polystyrene resins in bead form (one with a nominal surface area of 300 m(2)/g, the other with 750 m(2)/g) and an experimental proprietary molecular sieve with hydrophobic properties. Equilibrium adsorption isotherms for two of the sorbents were obtained at ambient temperature (21 degrees C) for ethanol-water solutions containing up to 12 wt. % ethanol. In addition, 40 degrees C isotherms were obtained for the polystyrene sorbents. Although different, the equilibrium isotherms for the sorbents indicated that ethanol could be preferentially sorbed from a dilute solution. Column breakthrough curves indicated very favorable kinetics. Desorption of the ethanol was readily effected with warm (60-80 degrees C), dry nitrogen.     

Power consumption of three impeller combinations in mixing Xanthan fermentation broths

Xanthan gum fermentation represents a good model for the study of the mixing of rheologically complex culture broths. Most of the previous work on power consumption dealt with ‘standard’, single impellers and used model fluids to simulate xanthan broths. This work describes the characterization of three dual-impeller combinations (D/T = 0·53) for the mixing of dehydrated—reconstituted fermentation broths of Xanthomonas campestris that had matched rheology to the actual broths. The bottom impeller was a Rushton turbine (RT) and the top impeller was another RT, a 45° pitched blade turbine (PT) or an A-310 Lightnin mixer (A310). The experiments were carried out in a tank of 0·0094 m³ working volume equipped with an air bearing dynamometer. The power was measured in a wide range of xanthan concentrations (5–40 kg m⁻³) in aerated (0·25, 0·5 and 1·0 vvm) and unaerated conditions. Unaerated power number (Po) vs. Reynolds number (Re) curves showed similar trends for the three combinations. Exponents close to −1 were obtained in the laminar region. A minimum in Po (Po_min) occurred at Re = 30–40, then increasing to a plateau value which was evident at Re> 200. In the transition region Po_min values were 4·3 (RT and RT), 3·6 (RT and PT) and 2·4 (RT and A310). The aerated power data for (RT and PT) and (RT and A-310) showed higher torque instabilities than the dual RT combinations at higher xanthan concentrations. The higher the xanthan concentrations, the higher the drop in power and the less important the effect of the aeration rate. Among the combinations tested, when using Rushton turbines, the well-mixed ‘cavern’ reached the tank wall (i.e., fluid motion was observed) at the lowest volumetric power input. High     

Salting-out extraction and crystallization of succinic acid from fermentation broths

In this study, salting-out extraction (SOE) and crystallization were combined to recover succinic acid from fermentation broths. Of the different SOE systems investigated, the system consisting of organic solvents and acidic salts appeared to be more favorable. A system using acetone and ammonium sulfate was investigated to determine the effect of phase composition and pH. The highest partition coefficient (8.64) and yield of succinic acid (90.05%) were obtained by a system composed of 30% (w/w) acetone and 20% (w/w) ammonium sulfate at a pH of 3.0. Additionally, 99.03% of cells, 90.82% of soluble proteins, and 94.89% of glucose could be simultaneously removed from the fermentation broths. Interestingly, nearly 40% of the pigment was removed using the single-step salting-out extraction process. The analysis of the effect of pH on salting-out extraction indicates that a pH lower than the pK of succinic acid is beneficial for the recovery of succinic acid in an SOE system. Crystallization was performed for the purification of succinic acid at 4 °C and pH 2.0. By combining salting-out extraction with crystallization, an identical total yield (65%) and a higher purity (97%) of succinic acid were obtained using a synthetic fermentation broth compared with the actual fermentation broth (65% and 91%, respectively).     

Reverse osmosis processing of organic model compounds and fermentation broths

Post-treatment of an anaerobic fermentation broth was evaluated using a 150 gal/day, single cartridge prototype reverse osmosis (RO) system. Baseline tests were conducted at 25 degrees C using six organic model compounds representing key species found in the fermentation broth: ethanol, butanol, acetic acid, oxalic acid, lactic acid, and butyric acid. Correlations of the rejection and recovery efficiencies for these organic species, individually and in simulated mixtures, were obtained as a function of feed pressure with and without recirculation of the retentate. The actual fermentation broth obtained from a continuous-flow biohydrogen process was treated by the RO system under the operating conditions similar to those used in the baseline tests, resulting in greater than 95% removal of total organic carbon. These results are encouraging and useful for further studies on the feasibility of incorporating the RO technology into an integrated and field deployable wastewater management and water recovery system.     

Application of ion exchange to purify acarbose from fermentation broths

Acarbose is conventionally used to reduce the insuline consumption of the diabetic patients. This compound is an oligosaccharide with the general formulae C25H43NO18 and obtained from fermentation processes by certain strains of Actinoplanes Utahensis. After the fermentation process, the acarbose has to be isolated from the fermentation broth where is accompanied of a large amount of substances, such as substrates, intermediate metabolites, proteins and different salts.

Four strong acid resins considering geliform and macroporous matrix types in aqueous and organic media have been tested in order to reach an easy and selective separation process. According to the experimental data, the Finex CS10GC (a gel strong cationic ion exchanger) presented the maximum acarbosa uptake and also the highest rate of ion exchange in water. The best behavior in non-aqueous media was observed with the Purolite CT151 (macroporous ion exchanger) but its maximum capacity of ion exchange was really lower than that exhibited by the Finex CS10GC resin in aqueous media. These results suggest that the acarbose removal from fermentation broths must be carried out in aqueous media to ensure the maximum usage of the resin uptake capacity. The results obtained provide a significant insight into the main equilibrium phenomena that takes place depending on the characteristics of the liquid phase. Finally, the elution of acarbose from the resin can be accomplished of a selectivity way by using a solution of 2.25N of HCl. The proposed separation method seems to be technically and economically feasible.     

Rheological characterization of xanthan fermentation broths and their reconstituted solutions

Two strains of X. campestris from a collection and a wild strain isolated from infected cabbage were cultured in a defined medium in a 25 dm³ fermenter. The rheological characteristics of the broth were measured using a Weissenberg rheogoniometer. The rheological behaviour over a wide range of shear rates could be described with the power law model and a yield stress was obtained using the Herschel-Bulkley equation. As expected, the broth's flow behaviour is highly influenced by gum concentration. In general, the rheological parameters during the fermentations varied within the following ranges: K (2–210 dyn-sⁿ/cm²), n (0.2–0.9), τ _y (0.02–50 dyn/ cm²), with strain 1459 reaching the maximum values as well as the highest gum concentration. Differences in flow behaviour could be associated with different molecular configurations of the polymers produced by the different strains. Heat treatment of the broths change the rheological characteristics, increasing K and decreasing n. Reconstituted solutions of all sterilized gums exhibited similar rheological characteristics and, at low concentrations, also similar to those of a commercial xanthan.