Automated fermentation equipment II. Computer–fermentor system

An expensive computer-operated system suitable for data collection and steady-state optimum control of fermentation processes is presented. With this system, minimum generation time has been determined as a function of temperature and pH in the turbidostat cultivation of a yeast strain. The applicability of the computer-fermentor system is also presented by the determination of the dynamic K_L a value.     

Ethanol fermentation in a continuous tower fermentor

A mutant of Saccharomyces cerevisiae, which forms large, multicellular flocs in liquid culture, rapidly fermented media containing high concentrations of glucose (100-180 g/L) in a continuous nonaerated tower fermentor at 30 degrees C. The fermentor operated continuously for seven months. Batch and tower fermentor data were fitted to a kinetic model incorporating linear ethanol inhibition and Monod dependence on glucose. Conversion, ethanol yield, and ethanol productivity were related to the apparent fermentation time for initial glucose concentrations of 130 and 180 g/L. Productivities of 8-12 g ethanol/L h were achieved through the yeast bed giving conversions exceeding 90% of the theoretical yield.     

A fermentor for study of sauerkraut fermentation

A fermentor was designed and constructed for study of the physical, microbiological, and chemical changes that occur during the sauerkraut fermentation. The fermentor has some essential features that include restriction in volume of the sauerkraut bed, construction of clear plastic to permit visual determination of liquid-level changes as a result of gas entrapment within the sauerkraut bed, and a gas-lift device for use in nitrogen purging of the fermenting brine. Fermentations exhibited two distinct stages, the first one gaseous and the second non-gaseous. The gaseous stage was characterized by rapid CO(2) and acid production due to growth by hetero-fermentative lactic acid bacteria with resultant gas entrapment within the sauerkraut bed and a rise in liquid level. Also, rapid disappearance of fructose and rapid appearance of mannitol occurred during this stage. The nongaseous stage was characterized by growth of homo-fermentative lactic acid bacteria with little or no CO(2) production and a gradual increase in lactic acid until all fermentable sugars were metabolized. Nitrogen purging appeared to offer several potential advantages, including a means for brine circulation, removal of CO(2) from the brine, and anaerobiosis to ensure retention of ascorbic acid, desirable color, and other oxygen-sensitive traits in sauerkraut.     

Ethanol fermentation in a yeast immobilized tubular fermentor

In this article, a mathematical model describing the kinetics of ethanol fermentation in a whole cell immobilized tubular fermentor is proposed. Experimental results show reasonable agreement with the proposed model. A procedure for treating the fermentation data for determining the ethanol inhibition constants k(1) and k(2) is described. The ethanol productivity of the immobilized cell fermentor is compared with those of traditional fermentors. Experimental studies indicate that with Saccharomyces cerevisiae (NRRL Y132) culture, ethanol productivity in the range 21.2-83.7 g ethanol L(-1) h(-1) at ethanol concentration of 76-60 g/L can be achieved. This is comparable to or higher than those reported in the literature for yeast. The product yield factor of 0.5 g ethanol/g glucose was obtained. The immobilized cell fermentor does not show washout at dilution rates of 7 h(-1) and shows good stability over a 650-h operating period.     

In vitro simulation of rabbit cecal fermentation in a semi- continuous flow fermentor. I. Role of food substrate pretreatment]

A Rusitec semi-continuous flow fermentor was used to study the influence of enzyme pretreatment of food substrates on the fermentation profile over a 2-week period following inoculation with rabbit caecal contents. Three types of substrate were examined: 1) homogenized commercial rabbit feed; 2) the solid remains of this feed after digestion with alpha-amylase for 24 h; and 3) substrate 2 digested for 4 h with pepsin (double enzyme treatment). One of a pair of nylon pouches containing 15 g substrate was replaced each day, thus producing a uniform 48-h fermentation. Fermentation of the untreated feed (1) for 5-6 days produced a fermentation profile quite different from that obtained in vivo in the rabbit caecum: propionic acid accounted for over 35% of total volatile fatty acid (VFA), and butyric acid for about 15%. Amylase digestion (2) gave a stable ferment profile closer to the in vivo profile, except that propionic and butyric acids were similar at 15% of total VFA. Digestion with both amylase and pepsin (3) produced a stable fermentation profile very close to the in vivo profile: C2 > 60%, C3 < 11% and 17% < C4 < 21%. The rate at which membrane constituents (acid detergent fibre, ADF) were lost in 48 h was similar to the digestibility coefficient measured in vivo by others for the same basic feed. Lastly, there was a high percentage (about 5%) of volatile C5 fatty acids; this could be due to the discontinuous fermentor input of one pouch per 24 h. Thus, feed pretreated with both amylase and pepsin simulates, in vitro, rabbit caecal fermentation in a semi-continuous Rusitec type fermentor.     

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.     

A continuous,multistage tower fermentor. I. Design and performance tests

The design of a continuous column fermentor with a multiple staging effect is described. The column is divided into four compartments by horizontal perforated plates and is provided with a central agitator shaft driving an impeller in each compartment. A tube at the center of each plate forms a liquid seal around the shaft and also acts as a “downcomer.” The fermentor is normally operated with counter-current flow of gas and medium. Fresh medium is added to the top stage and product is withdrawn from the bottom. The effect of plate and agitator design on fermentor performance was studied in terms of factor such as oxygen transfer rate, gas holdup, and interstage mixing. By proper choice of the design parameters, the fermentor was made to approximate a perfect four-stage cascade in terms of reactor performance. Preliminary experiments were performed with air-water systems, but a more realistic picture of fermentor performance was obtained in experience involving propagation of Escherichia coli. Data for business and substrate concentrations in each stage confirmed the staging effect of the apparatus. The fermentor operated in a stable manner for periods of more than two weeks.     

Solid-state fermentation of sweet sorghum to ethanol in a rotary-drum fermentor

Sugar compounds present in chopped solid-sweet sorghum particles were fermented to ethanol in a rotarydrum fermentor (RDF) using an ethanol tolerant yeast strain. The influence of rotational speed of the RDF on the rate of ethanol fermentation was investigated and compared with static flask experiments. The rate of ethanol formation decreased with increasing rotational speed. The maximum rate and extent of ethanol formation were ca. 3.1 g EtOH/L h (based on expressed juice volume) and ca. 9.6 g EtOH/100 g mash, respectively, at 1 rpm rotational speed.     

The optimization of Marasmius androsaceus submerged fermentation conditions in five-liter fermentor

Using desirability function, four indexes including mycelium dry weight, intracellular polysaccharide, adenosine and mannitol yield were uniformed into one expected value (Da) which further served as the assessment criteria. In our present study, Plackett–Burman design was applied to evaluate the effects of eight variables including initial pH, rotating speed, culture temperature, inoculum size, ventilation volume, culture time, inoculum age and loading volume on Da value during Marasmius androsaceus submerged fermentation via a five-liter fermentor. Culture time, initial pH and rotating speed were found to influence Da value significantly and were further optimized by Box–Behnken design. Results obtained from Box–Behnken design were analyzed by both response surface regression (Design-Expert.V8.0.6.1 software) and artificial neural network combining the genetic algorithm method (Matlab2012a software). After comparison, the optimum M. androsaceus submerged fermentation conditions via a five-liter fermentor were obtained as follows: initial pH of 6.14, rotating speed of 289.3 rpm, culture time of 6.285 days, culture temperature of 26 °C, inoculum size of 5%, ventilation volume of 200 L/h, inoculum age of 4 days, and loading volume of 3.5 L/5 L. The predicted Da value of the optimum model was 0.4884 and the average experimental Da value was 0.4760. The model possesses well fitness and predictive ability.     

Pilot plant fermentation of Streptomyces nigrificans in a mechanically agitated fermentor

Summary Optimal conditions for Streptomyces nigrificans growth, yields of glucose isomerase and protease activity were tested during fermentations in a mechanically agitated pilot plant fermentor, at two different impeller rotational speeds.     

Rapid ethanol fermentation of cellulose hydrolysate. II. Product and substrate inhibition and optimization of fermentor design

High concentrations of both ethanol and sugar in the fermentation broth inhibit the growth of yeast cells and the rate of product formation. Inhibitory effects of ethanol on the yeast strain Saccharomyces cerevisiae NRRL-Y-132 were studied in batch and continuous chemostat cultures. Growth was limited by either glucose or ethanol. Feed medium was supplemented with different ethanol concentrations. Ethanol was found to inhibit growth and the activity of yeast to produce ethanol in a noncompetitive manner. A linear kinetic pattern for growth and product formation was observed according to μ = μ_m (1 – P/P_m) and v = v_m (1 – P/P_m′), where μ_m is the maximum specific growth rate at P = 0 (hr^?1); P_m is the maximum specific product formation rate at P = 0 (hr^?1); P_m is the maximum ethanol concentration above which cells do not grow (g/liter); P_m′ is the maximum ethanol concentration above which cells do not produce ethanol (g/liter). Substrate inhibition studies were carried out using short-time experimental techniques under aerobic and anaerobic condition. The degree of substrate inhibition was found to be higher than that has been reported for ethanol fermentation of pure sugar. The kinetic relationships thus obtained were used to compute growth, substrate utilization, and alcohol production patterns and have been discussed with reference to batch and continuous fermentation of enzymatically produced bagasse hydrolysate.     

Scale-up of purine nucleoside fermentation from a shaking flask to a stirred-tank fermentor

Scaling-up purine nucleoside fermentation by a mutant strain of Bacillus subtilis from a shaking flask to a stirred-tank fermentor was attempted. The dimensions and the operating conditions of the stirred tank were determined in order to satisfy the optimum conditions of O₂ transfer and power consumption per unit volume for the shaking flask. When the purine nucleoside fermentation was carried out in the stirred-tank fermentor under these conditions, in which the temperature simulated that in the shaking flask, the total amount of purine nucleosides produced was almost the same as that in the shaking flask, but the accumulation ratio of guanosine to total nucleotides was different from that in the flask. Since urea could not be utilized so efficiently in the stirred-tank fermentor, the NH^p+ _f4 concentration and the pH of the culture broth were lower than those in the shaking-flask culture during fermentation. The activity of inosine monosphosphate dehydrogenase and the accumulation ratio were significantly affected by the NH^p+ _f4 concentration. When the pH of the stirred-tank culture was maintained at 6.9 by ammonia water to keep the NH^p+ _f4 level higher, the ratio was improved to the same level as that observed in the shaking-flask culture. The fermentation heat calculated from the shaking-flask data and its pattern of change were similar to those in the stirred-tank fermentor. Correspondence to: Y. Sumino     

Combined discrete particle and continuum model predicting solid-state fermentation in a drum fermentor

The development of mathematical models facilitates industrial (large-scale) application of solid-state fermentation (SSF). In this study, a two-phase model of a drum fermentor is developed that consists of a discrete particle model (solid phase) and a continuum model (gas phase). The continuum model describes the distribution of air in the bed injected via an aeration pipe. The discrete particle model describes the solid phase. In previous work, mixing during SSF was predicted with the discrete particle model, although mixing simulations were not carried out in the current work. Heat and mass transfer between the two phases and biomass growth were implemented in the two-phase model. Validation experiments were conducted in a 28-dm3 drum fermentor. In this fermentor, sufficient aeration was provided to control the temperatures near the optimum value for growth during the first 45-50 hours. Several simulations were also conducted for different fermentor scales. Forced aeration via a single pipe in the drum fermentors did not provide homogeneous cooling in the substrate bed. Due to large temperature gradients, biomass yield decreased severely with increasing size of the fermentor. Improvement of air distribution would be required to avoid the need for frequent mixing events, during which growth is hampered. From these results, it was concluded that the two-phase model developed is a powerful tool to investigate design and scale-up of aerated (mixed) SSF fermentors.     

Continuos IBE fermentation by immobilized growing Clostridium beijerinckii cells in a stirred-tank fermentor

The potential of continuous isopropanol-butanol-ethanol (IBE) fermentation by Ca-alginate-immobilized Clostridium beijerinckii cells in a continuous stirred-tank reactor is investigated. A mathematical model is presented to describe steady-state reactor performance. It appeared to be possible to use the biocatalyst particles repeatedly for successive fermentations (at least three times for a total duration of two months). Reactor productivity was 6-16 times higher than that of a batch fermentation (free cells), while the solvents yield was also increased. Measurements of substrate, product, and biomass concentrations were only partially in agreement with the model; however, a solid basis for further technological developement of the process has been laid.     

Optimization of monascus red pigment fermentation by regulating chitinase activity level in fermentor

Enhancement of monascus pigment production in a co-culture of Monascus sp. J101 with Saccharomyces cerevisiae was accomplished using chitinase in the feeding of a S. cerevisiae culture filtrate. Batch fermentations were carried out at constant chitinase activity levels of 0.8?U and 0.5?U after 24?h of cultivation. Monascus red pigment concentrations obtained at the end of fermentation were 182.4 OD units and 147.3 OD units, respectively, for chitinase activity levels of 0.8?U and 0.5?U. The cell mass was higher at 0.8?U than at 0.5?U, whereas pigment production per unit cell mass was higher at 0.5?U than at 0.8?U. Kinetic equations involving cell growth rate, cell activity and chitinase activity levels were analyzed. Cell activity was expressed as a function of mean cell age. The optimum time for reduction of the chitinase activity level from 0.8?U to 0.5?U for maximization of monascus pigment production was numerically estimated to be 60?h. Based on this result, a batch fermentation was performed where the chitinase activity level was maintained at 0.8?U from 24?h to 60?h, then the level was lowered to 0.5?U after 60?h. This scheme resulted in a monascus pigment production of 198.3 OD units.     

Citric acid production from glucose. I. Growth and excretion kinetics in a stirred fermentor

The growth and citric acid production kinetics of Saccharomycopsis lipolytica on glucose are investigated in an aerated stirred fermentor. Cellular growth first proceeds exponentially until exhaustion of ammonia in the fermentation medium. Cells then continue to grow at a reduced rate with a concomitant decrease in intracellular nitrogen content. Citric and isocitric acid production starts at the end of the growth phase. During about 80 hr excretion proceeds at a constant rate of 0.7 g/liter/hr for citric acid and 0.1 g/liter/hr for isocitric acid. The final citric and isocitric acid concentrations are 95 and 10g/liter, respectively. During acid excretion cellular respiration accounts for 60 and 35% of consumed oxygen and glucose. Both acid and CO₂ production rates follow a Michaelis–Menten-type dependence on oxygen concentration with Michaelis–Menten constants of 0.9 and 0.15 mg/liter for acid and CO₂ productions, respectively.     

In vitro simulation of rabbit cecal fermentation in a semi- continuous flow fermentor. II. Effect of inoculum type]

Two Rusitec fermentors were operated under identical conditions. One was seeded with an inoculum of rabbit caecal contents, and the other with bovine rumen contents. The fermentation substrate was rabbit feed that had been digested with amylase and pepsin. The substrate constituents (organic matter, OM and NDF) were lost in 48 h at a significantly higher rate in the presence of rumen inoculum (OM: +10%, NDF: +15%). The pHs of the 2 fermentors were similar at pH 6.6. The fermentors produced similar amounts of protein nitrogen per 24 h, after 6 d of adaptation. Volatile fatty acid production was slightly higher in the presence of rumen inoculum. The fermentor inoculated with rumen contents produced a higher percentage of propionic acid (25%) than of butyric acid (7%), while fermentation with rabbit caecal contents gave the opposite ratio (C3/C4 = 0.81). Consequently, only the rabbit caecal inoculum provided the fermentation profile characteristic of the species.