Elimination of oscillations in fermentation processes

Fermentation processes that are used to produce ethanol exhibit oscillatory behavior and for periods of time during these oscillations, ethanol production decreases substantially and there is considerable loss of residual substrates. There has been a considerable amount of work demonstrating oscillatory behavior in fermentation processes. The aim of this work is to demonstrate simple strategies to eliminate the oscillatory behavior in fermentation processes involving both Zymomonas mobilis and Saccharomyces cerevisiae. The oscillatory behavior is caused by the existence of Hopf bifurcations and it is demonstrated that very minor changes in the input conditions can eliminate the Hopf bifurcation points. In the case of Zymomonas mobilis, it is shown that the addition of a small amount of substrate and/or the key component of the biomass and/or product in the input stream causes the disappearance of the Hopf bifurcation points while in Saccharomyces cerevisiae fermentation process, a very minor increase in the input oxygen supply produces the same result. The aim of this work is not only to demonstrate the existence of the Hopf bifurcations in the fermentation problems, but also to provide strategies to eliminate them. © 2010 American Institute of Chemical Engineers AIChE J, 2011     

Ethanol fermentation by Z. mobilis ATCC10988 in repeated batch cultures

Repeated-batch fermentations of glucose to ethanol by Z. mobilis (ATCC 10988) were examined in two semi-defined nutrient media. The measurement of acid produced by the microorganism was used to study its transient fermentation characteristics. An examination of the inhibitory effect of ethanol in repeated-batch fermentations supports an earlier finding on the presence of a ‘slow-acting inhibitory mechanism’ at high ethanol concentration in Z. mobilis (ATCC 10988) cultures. The experimental results also suggest that both microbially produced and exogeneously added ethanol have similar inhibitory effects on the fermentation behavior of this bacterium.     

Direct ethanol production from dextran industrial waste water by Zymomonas mobilis

The direct production of ethanol from dextran industrial waste water was investigated by using Zymomonas mobilis via batch and semi-continuous fermentation mode. In batch fermentation, pretreated waste water (unsterilized and sterilized), pH value (3.8 and 6.0), and Mg²⁺ (with and without) was compared with OD₆₀₀, sugar and ethanol concentration. After 24 h fermentation, sugar in the dextran waste water was almost exhausted, and the amount of ethanol accumulated reached 24.33–29.92 g/l, which is nearly 99% of the theoretical yield of ethanol. Kinetic parameters of Z. mobilis in batch fermentation were also investigated. The raw dextran waste water was also used in semi-continuous fermentation. After 48 h fermentation, the production of ethanol was 28.65 g/l. These results indicated that dextran waste water may be used as a candidate substrate and Z. mobilis could convert the raw material into ethanol directly.     

Bifurcations of Propellant Burning Rate at Oscillatory Pressure

A new phenomenon, the disparity between pressure and propellant burning rate frequencies, has revealed in numerical studies of propellant burning rate response to oscillatory pressure. As is clear from the linear approximation, under small pressure amplitudes, h, pressure and propellant burning rate oscillations occur with equal period T (T‐solution). In the paper, however, it is shown that at a certain critical value of the parameter h the system in hand undergoes a bifurcation so that the T‐solution converts to oscillations with period 2T (2T‐solution). When the bifurcation parameter h increases, the subsequent behavior of the system becomes complicated. It is obtained a sequence of period doubling to 4T‐solution and 8T‐solution. Beyond a certain value of the bifurcation parameter h an apparently fully chaotic solution is found. These effects undoubtedly should be taken into account in studies of oscillatory processes in combustion chambers.     

Miscibility and crystallization behavior of poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate)/poly(vinyl acetate) blends

The miscibility and crystallization behavior of poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate) (P(HB‐co‐HV))/poly(vinyl acetate) (PVAc) blends have been investigated by differential scanning calorimetry (DSC) and polarized optical microscopy (POM). It was found that P(HB‐co‐HV)/PVAc blends were miscible in the melt over the whole compositions. Thus the blend exhibited a single glass transition temperature (T_g), which increased with increasing PVAc composition. The spherulitic morphologies of P(HB‐co‐HV)/PVAc blends indicated that the PVAc was predominantly segregated into P(HB‐co‐HV) interlamellar or interfibrillar regions during P(HB‐co‐HV) crystallization because of the volume‐filled spherulites. As to the crystallization kinetics study, it was found that the overall crystallization and crystal growth rates decreased with the addition of PVAc. The kinetics retardation was primarily attributed to the reduction of chain mobility and dilution of P(HB‐co‐HV) upon mixing with higher T_g PVAc. The overall crystallization rate was predominantly governed by the spherulitic growth rate and promoted by the samples treated with the quenched state because of the higher nucleation density. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100: 980–988, 2006     

Bifurcation analysis of two continuous bioreactors operated in series with recycle

In this paper, bifurcation analysis has been carried out for two continuous bioreactors operated in series with recycle from the second reactor. The existence of multiplicity of steady states is analyzed by considering Contois growth kinetics in the process model. It was observed that there exist two possible steady states of which one is trivial (wash out condition). Stability analysis is carried out to determine the stability of these steady states and it was observed that both these steady states are unstable in nature. Bifurcation analysis has been carried out for substrate and biomass concentration with dilution rate as the bifurcation parameter. Effect of recycle ratio, substrate separation factor and biomass separation factor is studied and analyzed. It was observed that Hopf bifurcation occurs at a dilution rate of 1.0208 with purely imaginary Eigen values which showed that sustained oscillatory behavior exists in the substrate concentration of the second reactor. The significance of different bifurcation points and the operating conditions by considering biomass and substrate concentrations in each reactor is studied and it was observed that the bioreactors need to be operated at intermediate dilution rates to obtain improved conversion and yield.     

Dynamic and chaotic behavior of periodically forced fermentors for bioethanol production

In this preliminary paper, the dynamic responses of bioethanol fermentors to sinusoidal periodic perturbations of the feed concentration have been investigated. It is shown that the steady state autonomous model of the unforced fermentor can be reduced analytically to a cubic polynomial. This analytical form enables to implement the global bifurcation analysis to divide the parameter space into regions with different number of steady state solutions; moreover, the discriminant of the cubic polynomial is used for the analysis of the nature of the number of the steady state solutions. The bifurcation analysis shows that the locus of the fold points is equivalent to the locus of the discriminant of the cubic polynomial in the (D, C_so) plane. It has been shown that the periodic operation of the autonomous fermentor gives higher average ethanol concentration than the corresponding unstable steady state. The investigation reveals that the centre of forcing has significant effect on the dynamic response of the periodically forced fermentors. Chaotic behavior is developed when the centre of forcing is relatively close to a homoclinic infinite period orbit, while quasi-periodicity is developed when the centre of forcing is in the neighborhood of a Hopf bifurcation point. The system shows interesting phase planes at certain forcing amplitudes. It has been shown that within the parameters range studied the best policy for production of bioethanol is to operate the forced fermentors in the frequency locking regions at small forcing amplitudes.     

Exploration and exploitation of bifurcation/chaotic behavior of a continuous fermentor for the production of ethanol

A four-dimensional model for the anaerobic fermentation process, developed and used earlier to simulate the oscillatory behavior of an experimental continuous stirred tank fermentor is utilized in the present investigation to explore the static/dynamic bifurcation and chaotic behavior of this fermentor, which is shown to be quite rich. The present investigation is a prelude to the experimental exploration of bifurcation and chaos in a membrane fermentor.Dynamic bifurcation (periodic attractors) as well as period doubling sequences leading to different types of periodic and chaotic attractors have been uncovered. It is fundamentally and practically important to discover the fact that in some cases, periodic and chaotic attractors have higher ethanol yield and production rate than the corresponding steady states.     

Improvement of ethanol production in Saccharomyces cerevisiae by hetero‐expression of GAPN and FPS1 deletion

BACKGROUND: During anaerobic bioethanol fermentation of Saccharomyces cerevisiae, the main byproduct glycerol is essential to regulate redox balance (reoxidize NADH to NAD⁺), which is necessary to maintain cell growth and fermentation. Hetero‐expression of a NADP⁺‐dependent glyceraldehydes‐3‐phosphate dehydrogenase (GAPN) [EC.1.2.1.9] in S. cerevisiae could redirect the carbon flux from glycerol to ethanol involving a net oxidation of NADH. The present study investigates whether combination of GAPN hetero‐expression and glycerol exporter Fps1p disruption would result in less glycerol and more ethanol production without affecting growth rate during anaerobic fermentations. RESULTS: The results of anaerobic fermentations showed that the fps1Δ mutant with GAPN (named 4FG) produced 21.47% less glycerol and 9.18% more ethanol compared with a parental strain with a control plasmid, while the rates of growth and fermentation were not changed. Moreover, the engineered strain 4FG yielded less glycerol and acetic acid, and more ethanol than the control, fps1Δ mutant or with GAPN only. CONCLUSIONS: During anaerobic fermentations, hetero‐expression of GAPN restored the reduced grow rate of the fps1Δ mutant, and led to less byproducts and more ethanol production. This combination strategy could be used to modulate glycerol metabolism and optimize the anaerobic fermentation of S. cerevisiae. Copyright © 2011 Society of Chemical Industry     

THE EFFECT OF MACROMIXING ON OSCILLATORY BEHAVIOR IN RECOMBINANT FERMENTATION IN A CONTINUOUS STIRRED TANK REACTOR

A general model is presented for recombinant fermentation in a chemostat in which micromixing may be imperfect. Using the production of tryptophan synthetase as an example and the dilution rate, D, as the bifurcation parameter, conditions are first derived for Hopf bifurcation in a perfectly mixed chemostat. The bifurcation surfaces in three-dimensional space are such that oscillatory solutions exist above the surfaces and washout conditions below. A similar analysis is done for imperfect mixing. The bifurcation region is a concave hyperboloid in the [D,f, n]-space, where f and n ; are model parameters. It is argued that poor macromixing reduces the range of D over which bifurcation to periodic solutions is possible.     

Miscibility and crystallization kinetics of poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate)/ poly(methyl methacrylate) blends

The miscibility and crystallization kinetics of the blends of random poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate) [P(HB‐co‐HV)] copolymer and poly(methyl methacrylate) (PMMA) were investigated by differential scanning calorimetry (DSC) and polarized optical microscopy (POM). It was found that P(HB‐co‐HV)/PMMA blends were miscible in the melt. Thus the single glass‐transition temperature (T_g) of the blends within the whole composition range suggests that P(HB‐co‐HV) and PMMA were totally miscible for the miscible blends. The equilibrium melting point (T°_m) of P(HB‐co‐HV) in the P(HB‐co‐HV)/PMMA blends decreased with increasing PMMA. The T°_m depression supports the miscibility of the blends. With respect to the results of crystallization kinetics, it was found that both the spherulitic growth rate and the overall crystallization rate decreased with the addition of PMMA. The kinetics retardation was attributed to the decrease in P(HB‐co‐HV) molecular mobility and dilution of P(HB‐co‐HV) concentration resulting from the addition of PMMA, which has a higher T_g. According to secondary nucleation theory, the kinetics of spherulitic crystallization of P(HB‐co‐HV) in the blends was analyzed in the studied temperature range. The crystallizations of P(HB‐co‐HV) in P(HB‐co‐HV)/PMMA blends were assigned to n = 4, regime III growth process. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 91: 3595–3603, 2004     

Stabilized production of highly concentrated bioethanol from fermentation broths by Zymomonas mobilis by pervaporation using silicone rubber‐coated silicalite membranes

Since pervaporation performance of ethanol‐permselective silicalite membrane, which is an aluminum‐free hydrophobic zeolite, in the separation of fermentation broths by yeast are negatively affected by succinic acid, the potential of pervaporation using silicone rubber‐coated silicalite membranes of ethanol fermentation broths, not containing succinic acid, by Zymomonas mobilis was investigated for the reliable production of concentrated bioethanol. In the separation of fermentation broths, the pervaporation performance was influenced by nutrients used for the preparation of fermentation broths. In the separation of a broth prepared with yeast extract, pervaporation performance was greatly compromised by accumulation of a substance(s) having an ultraviolet absorption maximum at approximately 260 nm not only in total flux, but also in permeate ethanol concentration compared to the separation of binary ethanol/water mixtures. When supplying a prepared broth with corn steep liquor without the accumulation of a substance(s) having an ultraviolet absorption maximum at approximately 260 nm, the permeate ethanol concentration did not decrease. Treating the prepared broth with activated carbon was effective in restraining the decrease in total flux. Pervaporation performance is also deteriorated by the adsorption of lactic acid contained in corn steep liquor onto the silicalite crystals. In the separation of ternary mixtures of ethanol/water/lactic acid, accomplished by adjusting the ternary mixtures to pH > 5, more than 90% of the permeation flux in the separation of binary ethanol/water mixtures was obtained, and the permeate ethanol concentration was comparable to that obtained in the separation of binary mixtures. For stably performing pervaporation, it is important to prepare ethanol fermentation broths by Zymomonas mobilis in which lactic acid concentration is as low as possible. Copyright © 2007 Society of Chemical Industry     

同时进行发酵和分离的CO_2气提、活性炭吸附乙醇发酵动力学研究——(二)游离细胞连续发酵

本文研究了游离细胞连续发酵时应用 CO_2气提、活性炭吸附的乙醇发酵和分离动力学,考察了稀释率和进口葡萄糖浓度对发酵速率的影响,提出了包括 CO_2汽提乙醇速率的发酵动力学模型,研究了连续发酵时的稳态和动态过程,结果表明,CO_2气提有助于减少乙醇对酵母的抑制作用,发酵动力学模型可以较好地关联稳态时的发酵动力学数据,从中获得的模型参数可以用于预测动态过程中底物、微生物及产物的变化规律。对于所研究的过程,存在着最佳稀释率和进口糖浓度以获得最大发酵速率。     

Kinetics of Bioethanol Production Employing Mono‐ and Co‐Cultures of Saccharomyces cerevisiae and Pichia stipitis

The kinetics of bioethanol production using mono‐ and co‐cultures of Saccharomyces cerevisiae and Pichia stipitis with glucose, xylose, and glucose‐xylose sugar mixtures were investigated. A MATLAB® program was formulated for simulation of experimental results in order to get predicted values of ethanol production and sugar consumption and for kinetic parameter estimation. Kinetic parameters implied less extent of substrate and/or product inhibition when the co‐culture scheme of immobilized S. cerevisiae and free P. stipitis was employed for fermentation of mixed sugars. In addition, a high ethanol yield was achieved by applying this co‐culture strategy to wheat straw hydrolysates.     

Platelet‐activating factor detection,metabolism, and inhibitors in the ethanologenic bacterium Zymomonas mobilis

Platelet‐activating factor (PAF) is a signaling phospholipid with a significant physiological role in multicellular and unicellular organisms, including fermentative organisms such as yeast. Zymomonas mobilis is an ethanologenic α‐proteobacterium currently studied for bioethanol production. In order to examine the presence of PAF and/or PAF inhibitors in Z. mobilis, a new one‐step high performance liquid chromatography (HPLC) separation procedure of total lipids was performed, using a C8 reversed‐phase semi‐preparative column. According to this method and to bioassays based on washed rabbit platelet aggregation, two lipid molecules with PAF‐like activity and same retention times as those of standard PAF were detected; electron‐spray ionization MS and MS/MS analysis revealed that they share similar structure with 16:0 and 18:0 PAF. Furthermore, other lipids extracted from Z. mobilis were found to exhibit a potent anti‐PAF activity. Enzyme activities indicative of key PAF biosynthetic enzymes, such as dithiothreitol‐insensitive cholinephosphotransferase (PAF‐CPT) and lyso‐PAF acetyltransferase were detected in Z. mobilis homogenates. As for PAF degradation, activity similar to that of PAF acetylhydrolase was also discovered. Overall, the presence of PAF, PAF‐specific inhibitors, and enzyme activities relating to PAF metabolism, suggests that PAF may play an intrinsic role in this biotechnological organism. Practical applications: Z. mobilis is a platform microorganism for bioethanol production and a potential source of high‐value chemicals of interest to the food and healthcare industries. Further investigation of PAF's role is bound to affect applications involving this and other biotechnological organisms. The finding that Z. mobilis lipids exhibit potent anti‐PAF activity opens up prospects for their identification, overproduction and pharmaceutical use. The presented HPLC method for lipid fractionation accomplishes a one‐step separation of lipids from dense samples, which may be successfully employed to other lipid‐rich sources such as blood.     

Metabolic responses of microorganisms growing on inhibitory substrates in nonsteady state culture

The dynamics of metabolic activity of Pseudomonas putida cultured on phenol in a nonsteady state, resulting from simultaneous step increases (or decreases) in dilution rate and feed phenol concentration, were studied. Metabolic activity was determined by the uptake rate, defined as the mass of phenol degraded by a unit mass of bacteria in unit time. A minimum duration of nonsteady state was necessary to induce the metabolic pathway. The induction time for Pseudomonas putida cultured on phenol was approximately 30 min, irrespective of the type of disturbance applied. The Monod-Haldane equation was applied to fit the experimental values of uptake rate and the best kinetic parameters were calculated by the nonlinear least squares technique. Values of the kinetic parameters differed both for each parameter and type of disturbance investigated. For simultaneous step increases in dilution rate and feed phenol concentration, the value of u_m in the transient state first increased monotonically approximately four times and then decreased monotonically to the new steady state value after three residence times. Values of K_s and K_i attained those at the new steady state after three residence times, the highest being approximately three times greater than those of the steady state prior to disturbance. For simultaneous step decreases in D and S_O, values of u_m, K_s and K_i reached those at the new steady state values after four residence times.     

Ethanol-Herstellung mit Bakterien

Ethanol production with bacteria . Strains of Saccharomyces cerevisiae have mostly been used for the production of ethanol from sugar by yeasts. Recently it was shown that the bacterium Zymomonas mobilis has some advantages compared to yeast for the production of industrial alcohol. Compared to traditional yeast fermentation, ethanol yield is about 5% higher than with yeast, since less sugar is incorporated into cell material by this bacterium. Like yeast, Zymomonas mobilis has remarkably high ethanol tolerance which enables the bacterium to produce ethanol concentrations of more than 13 vol.-% from sugar solutions of appropriate concentration. Investigations of the spectrum of lipids present have shown that this bacterium contains large quantities of hopanoids which are presumably of significance for the stabilization of cell membranes in the presence of ethanol. Since the cost of the sugar greatly influences the profitability fraction formed in the production of glucose syrup from wheat flour was investigated. It was shown that after enzymatic saccharification of this waste starch the glucose was efficiently fermented to ethanol by Zymomonas mobilis. It is planned to broaden the substrate spectrum of Zymomonas mobilis by gene cloning techniques so that in future pentoses, e. g. xylose or arabinose, can also be fermented to ethanol by this organism.     

A study on the ethanol production by immobilized cells ofZymomonas mobilis

Cells ofZymomonas mobilis immobilized in Ca-alginate matrix were used for ethanol production under various conditions. Immobilized cells showed broad optimum pH profile and their operational optimum temperature shifted from 30‡C to 40‡C upon immobilization. As reportedlyZ. mobilis did get the substrate inhibition by glucose, but at high concentration level of glucose the reduction of activity for ethanol production was less severe than that for yeast. The used beads of the immobilizedZ. mobilis were reactivated by incubating them in the activation medium. The increase in cell number and the enhancement of the specific activity per each cell are considered the two major factors responsible for the overall activation. A packed-bed reactor with the feed glucose concentration of 20% (W/V) gave an ethanol productivity as high as 33.0 g/l.hr at the flow rate of 58.5 ml/hr. A comparison between the experimental results from the real packed-bed reactor and the simulation results of an ideal case showed a two-fold inferior performance by the real reactor and this is at least partly attributed to the CO₂ gas effect.     

Production of L‐lactic acid with very high gravity distillery wastewater from ethanol fermentation by a newly isolated Enterococcus hawaiiensis

BACKGROUND: A great amount of wastewater with high contents of chemical oxygen demand (COD) are produced by ethanol production. It would be useful to utilize distillery wastewater to produce L‐lactic acid, which could be a high additional value byproduct of ethanol production. The fermentation process of L‐lactic acid production by a newly isolated Enterococcus hawaiiensis CICIM‐CU B0114 is reported for the first time. RESULTS: The strain produced 56 g L^?1 of L‐lactic acid after cultivation for 48 h in optimized medium consisting of (g L^?1) 80 glucose, 10 peptone, 10 yeast extract, 1.5 Na₂HPO₄ and 0.2 MgSO₄. E. hawaiiensis CICIM‐CU B0114 was isolated and purified by subculture for growing and producing L‐lactic acid in distillery wastewater of very high gravity (VHG) from ethanol fermentation. L‐lactic acid fermentation was further studied with distillery wastewater substrate in 7 L and 15 L fermentors. The results showed that L‐lactic acid concentrations of 52 g L^?1 and 68 g L^?1 was achieved in 7 L and 15 L fermentors with the initial sugar concentrations of 67 g L^?1 and 87 g L^?1, respectively. CONCLUSION: The production of L‐lactic acid by the newly isolated E. hawaiiensis CICIM‐CU B0114 was carried out and the fermentation medium was optimized by orthogonal experimental design. This new strain holds the promise of L‐lactic acid production utilizing distillery wastewater from VHG ethanol fermentation. Copyright © 2010 Society of Chemical Industry     

DYNAMIC ANALYSIS OF GAS STRIPPING DURING ETHANOL FERMENTATION IN CSTR

1 INTRODUCTIONInhibition phenomena have frequently been observed in microbial reactions.When thesubstrate concentration in the broth is high enough,the contour curve of the specificgrowth rate of cells is no longer monotonically increasing in a substrate-product phaseplane.The system exhibits a substrate inhibition.Some supersession products can affectthe growth and the yield of the cells as well as the viability of the microorganisms.Forexample,in the case of using Saccharomyces,the ethanol product inhibits the growth