Tower fermentation using Zymomonas mobilis for ethanol production

Summary Flocculation was induced in a pure strain of the bacteria Zymomonas mobilis. When fermenting glucose to ethanol, cell densities of up to 40g/l were achieved and sustained in a 0.92 litre tower fermenter with dilution rates of up to 2.3 hr^-1. A maximum productivity of 100g EtOH/l/hr with 98% conversion of the 105g/l glucose feed was achieved. The limitation to performance with increase in throughput arose from incomplete fermentation of the feed glucose, rather than washout of the flocculated bacteria.     

Intracellular ethanol and cell viability ofZymomonas mobilis during fed-batch alcoholic fermentation

Summary The effect of intracellular as well extracellular ethanol concentration on the viability ofZymomonas mobilis during a fed-batch fermentation was examined. The cells retained their viability until ethanol attained 69.5 ± 1.55 and 69 ± 1.6g/l for respectively, extracellular and intracellular values.Z. mobilis does not therefore accumulate ethanol in the cells. The number of dead cells increased after exposure to ethanol. The maximal efficiency of the fermentation was 95.5% (3.82 mol of ethanol-mol sucrose).

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Contenu intra-cellulaire en éthanol et viabilité des cellules de Zymomonas mobilis pendant une fermentation éthanolique en milieu non renouvelé à alimentation étagée

Résumé On a examiné l'effet de la concentration en éthanol tant inta- que extra-cellulaire sur la viabilité deZymomonas mobilis pendant une fermentation en milieu non renouvelé à alimentation étagée. Les cellules ont gardé leur viabilité jusqu'à ce que la concentration atteigne 69.5 ± 1.55 et 69 ±a 1.6 g/l respectivement pour les valeurs extra- et intra-cellulaires. Il en résulte queZymomonas mobilis n'accumule pas l'éthanol dans les cellules. Le nombre de cellules mortes augmentait après l'exposition à l'éthanol. L'efficience maximum de la fermentation était de 95.5% (3.82 mol d'éthanol/mol de sucrose).

     

Vacuum fermentation for ethanol production using strains of Zymomonas mobilis

Summary Using strains of Z.mobilis, a vacuum fermentation system has been evaluated. The system was designed with the fermentor at atmospheric pressure and an external vacuum vessel (50 mm Hg). Sequential operation of the vacuum vessel was under microprocessor control. The use of Z.mobilis together with the two-stage design of the vacuum system has been found to overcome the problems of oxygen addition and the possibility of contamination reported previously for vacuum fermentations with yeasts. The productivity of 85 g/1/h found in the continuous cell recycle experiments was similar to that reported previously for a strain of S.cerevisiae.     

Aeration alleviates ethanol inhibition and glycerol production during fed-batch ethanol fermentation

In this study, we investigated the effects of aeration on ethanol inhibition and glycerol production during fed-batch ethanol fermentation. When aeration was conducted at 0.13, 0.33, and 0.8 vvm, the ethanol productivity, specific ethanol production rate, and ethanol yield in the presence of greater than 100 g/L of ethanol were higher than when aeration was not conducted. In addition, estimation of the parameters (α and β) in a model equation of ethanol inhibition kinetics indicated that aeration alleviated ethanol inhibition against the specific growth rate and the specific ethanol production rate. Specifically, when aeration was conducted, the glycerol yield and specific glycerol production rate decreased approximately 50 and 70%, respectively. Finally, the results of this study indicated that aeration during fed-batch ethanol fermentation may improve the ethanol concentration in the final culture broth, as well as the ethanol productivity.     

Ethanol production by Zymomonas mobilis in fed-batch fermentations

The production of ethanol by Zymomonas mobilis NRRL B-4286 was studied in fed-batch cultures. Initial percent (w/v) glucose, rate of feed, and quantity of 50%; (w/v) glucose feed were varied. Glucose inhibition of growth rate occurred at concentrations greater than 8% (w/) Feed was begun after 4 h incubation. Feed volume was ca. 36%; of starting batch volume to get ca. 10%; (w/v) ethanol at harvest. The range of feed rates studied varied from 16%; batch volume/h (glucose concentration increased to an inhibitory level) to 4%; batch volume/h (glucose concentration dropped rapidly to zero and was limiting). Increasing feed volume to 46%; of starting volume at the best feed rate (ca. 10%; feed volume/h) increased final ethanol concentration to 11.3%; (w/v). However, the resultant increase in fermentation time from ca. 21 to 29 h decreased ethanol volumetric productivity from 5.2 to 4.6 g/L h.     

Ethanol production from biomass by repetitive solid-state fed-batch fermentation with continuous recovery of ethanol

To save cost and input energy for bioethanol production, a consolidated continuous solid-state fermentation system composed of a rotating drum reactor, a humidifier, and a condenser was developed. Biomass, saccharifying enzymes, yeast, and a minimum amount of water are introduced into the system. Ethanol produced by simultaneous saccharification and fermentation is continuously recovered as vapor from the headspace of the reactor, while the humidifier compensates for the water loss. From raw corn starch as a biomass model, 95 ± 3, 226 ± 9, 458 ± 26, and 509 ± 64 g l⁻¹ of ethanol solutions were recovered continuously when the ethanol content in reactor was controlled at 10–20, 30–50, 50–70 and 75–85 g kg-mixture⁻¹, respectively. The residue showed a lesser volume and higher solid content than that obtained by conventional liquid fermentation. The cost and energy for intensive waste water treatment are decreased, and the continuous fermentation enabled the sustainability of enzyme activity and yeast in the system.     

Optimal nonsingular control of fed-batch fermentation

Presented is a new simple method for multidimensional optimization of fed-batch fermentations based on the use of the orthogonal collocation technique. Considered is the problem of determination of optimal programs for fermentor temperature, substrate concentration in feed, feeding profile, and process duration. By reformulation of the state and control variables is obtained a nonsingular form of the optimization problem which has considerable advantage over the singular case since a complicated procedure for determination of switching times for feeding is avoided. The approximation of the state variables by Lagrange polynomials enables simple incorporation of split boundary conditions in the approximation, and the use of orthogonal collocations provides stability for integration of state and costate variables. The interpolation points are selected to obtain highest accuracy for approximation of the objective functional by the Radau-Lobatto formula. The control variables are determined by optimization of the Hamiltonian at the collocation points with the DFP method. Constraints are imposed on state and control variables.The method is applied for a homogeneous model of fermentation with volume, substrate, biomass, and product concentrations as the state variables. Computer study shows considerable simplicity of the method, its high accuracy for low order of approximation, and efficient convergence.     

分批补料对木糖发酵生产乙醇的影响

以树干毕赤酵母为发酵菌种,纯木糖为发酵底物,通过分批补料来提高糖利用率以及乙醇得率。结果表明,在24h内,最佳初始木糖浓度为80g/L,在28h的发酵周期中,可以将木糖浓度提高至90g/L,在32h发酵周期内可以将木糖浓度提高至100g/L。通过分批补料,乙醇浓度得到明显提高。当总糖浓度分别为80g/L、90g/L时,24h发酵周期内,分批补料次数以1次为宜,乙醇浓度分别达30.95g/L、32.60g/L,相比于不补料即一次性投料,乙醇浓度分别提高了9.36%、9.18%。总糖浓度100g/L,28h发酵周期内,补料2次效果最佳,乙醇浓度达37.49g/L,比一次性投料下提高了10.36%,较一次性投料达到相同发酵效果缩短了4h。     

Optimization of fed-batch fermentation for xylitol production by Candida tropicalis

Xylitol, a functional sweetener, was produced from xylose by biological conversion using Candida tropicalis ATCC 13803. Based on a two-substrate fermentation using glucose for cell growth and xylose for xylitol production, fed-batch fermentations were undertaken to increase the final xylitol concentration. The effects of xylose and xylitol on xylitol production rate were studied to determine the optimum concentrations for fed-batch fermentation. Xylose concentration in the medium (100 g l⁻¹) and less than 200 g l⁻¹ total xylose plus xylitol concentration were determined as optimum for maximum xylitol production rate and xylitol yield. Increasing the concentrations of xylose and xylitol decreased the rate and yield of xylitol production and the specific cell growth rate, probably because of an increase in osmotic stress that would interfere with xylose transport, xylitol flux to secretion to cell metabolism. The feeding rate of xylose solution during the fed-batch mode of operation was determined by using the mass balance equations and kinetic parameters involved in the equations in order to increase final xylitol concentration without affecting xylitol and productivity. The optimized fed-batch fermentation resulted in 187 g l⁻¹ xylitol concentration, 0.75 g xylitol g xylose⁻¹ xylitol yield and 3.9 g xylitol l⁻¹ h⁻¹ volumetric productivity. Journal of Industrial Microbiology & Biotechnology (2002) 29, 16–19 doi:10.1038/sj.jim.7000257 Received 15 October 2001/ Accepted in revised form 30 March 2002     

Optimized nikkomycin production by fed-batch and continuous fermentation

We performed fed-batch and continuous fermentations to extend the time of maximal nikkomycin production by Streptomyces tendae Tü 901/S 2566. This was achieved by the fed-batch culture technique. Furthermore, high productivity was obtained at slow growth rates in a continuous fermentation process. Different dilution rates with and without carbon limitation were done and the results were compared.Correspondence to : T. Schüz     

High efficiency ethanol fermentation by entrapment of Zymomonas mobilis into LentiKats

AIMS: To examine the potential of Zymomonas mobilis entrapped into polyvinylalcohol (PVA) lens-shaped immobilizates in batch and continuous ethanol production. METHODS AND RESULTS: Cells, free or immobilized in PVA hydrogel-based lens-shaped immobilizates - LentiKats, were cultivated on glucose medium in a 1 l bioreactor. In comparison with free cell cultivation, volumetric productivity of immobilized batch culture was nine times higher (43.6 g l(-1) h(-1)). The continuously operated system did not improve the efficiency (volumetric productivity of the immobilized cells 30.7 g l(-1) h(-1)). CONCLUSIONS: We demonstrated Z. mobilis capability, entrapped into LentiKats, in the cost-efficient batch system of ethanol production. SIGNIFICANCE AND IMPACT OF THE STUDY: The results reported here emphasize the potential of bacteria in combination with suitable fermentation technology in industrial scale. The innovation compared with traditional systems is characterized by excellent long-term stability, high volumetric productivity and other technological advantages.     

Continuous ethanol production by immobilized cells of Zymomonas mobilis

Summary Studies have been carried out with a highly productive strain of Zymomonas mobilis in an immobilized cell reactor using both Ca alginate and -carrageenan as supporting matrices. Productivities above 50 g/l/h have been found at ethanol concentrations in excess of 60 g/l. With immobilized cells of Z. mobilis, there was a decline of approximately 30s% in activity after 800 h operation.     

Long-term repeated fed-batch ethanol fermentation in aerated condition

In this study, we attempted to assess the process stability of long-term fed-batch ethanol fermentation in the absence and presence of aeration (0.33 vvm). To examine the effect of aeration, a long-term repeated fed-batch operation was conducted for 396 h to mimic a long-term industrial bioethanol production process. In this long-term repeated fed-batch ethanol fermentation experiments, withdrawal-fill operation were conducted every 36 h for 10 repeat cycles. The whole operation was stably sustained in a quasi-steady state. The average maximal cell concentration and the average maximal ethanol production during operation were increased by 81.63 and 12.12%, respectively, when aeration was used. In addition, since aeration was carried out, the average ethanol yield slightly decreased by 4.03% and the average specific ethanol production rate decreased by 46.75% during operation. However, the average ethanol productivity increased by 17.53% when aeration was carried out. After 396 h of long-term repeated fed-batch ethanol fermentation, 1,908.9 g of ethanol was cumulatively produced when aeration was used, which was 12.47%, higher than when aeration was not used (1,697.2 g). Meanwhile, glycerol production was greatly decreased during long-term repeated fed-batch ethanol fermentation, in which the glycerol concentration in the culture broth decreased from about 34∼15 g/L. Thus, we can conclude that cell growth was greatly improved by overcoming ethanol inhibition and glycerol production was remarkably decreased when aeration was carried out, although aeration in ethanol fermentation decreased the specific ethanol production rate and ethanol yield.     

Semicontinuous diffusion fermentation of fodder beets for fuel ethanol and cubed protein feed production

A novel, semicontinuous diffusion fermentation system was used to produce fuel ethanol and a cubed protein feed (CPF) from fodder beets at an intermediate scale. In the process, fodder beet cubes were augered diagonally upward against a flow of 0.26N H(2)SO(4) and yeast in a tubular fermentor. Exiting one end of the fermentor was CPF, while fermented beer [6-9% (v/v) ethanol] exited the other end. Retention times for beer and CPF were 264 and 72 h, respectively. Contamination was controlled by maintaining the fermentation pH between 2.1 and 2.6 using H(2)SO(4). Production costs for a greatly scaled-up (times 1400) conceptual version of this system (using a continuous rather than a semicontinuous processing mode) were projected by calculation to be $0.529/L for 95% ethanol (net of a $0.112/L credit for CPF). The calculated energy balance (energy output-energy input ratio) was estimated to be 3.04. In calculating the energy balance, the output energy of the CPF and input energy for growing the fodder beets were not included. A design for the scaled-up plant is provided.     

Bacterial cellulose production by fed-batch fermentation in molasses medium

Batch and fed-batch fermentations for bacterial cellulose (BC) production using molasses as a carbon source by Acetobacter xylinum BPR2001 were carried out in a jar fermentor. For improvement of BC production, molasses was subjected to H2SO4-heat treatment. The maximum BC concentration by this treated molasses increased 76%, and the specific growth rate increased 2-fold compared with that by untreated molasses. In batch fermentation, when the initial sugar concentrations of H2SO4-heat-treated molasses were varied from 20 to 70 g/L, the highest value of maximum BC concentration of 5.3 g/L was observed at 20 g/L. BC production in intermittent fed-batch (IFB) fermentation was conducted referring to the data in batch fermentation, and the highest BC production of 7.82 g/L was obtained when 0.2 L of molasses medium was added five times. When continuous fed-batch (CFB) fermentations were conducted, maximum BC concentration was obtained with a feeding rate of 6.3 g-sugar/h, which was derived from the optimal IFB experiment.     

Continuous ethanol production by a flocculent strain of Zymomonas mobilis

Summary Cell retention and ethanol production using the flocculent bacterium Zymomonas mobilis NRRL B-12526 were studied in three bioreactor configurations. The flocculent growth characteristic of this strain and a special reactor design were combined to achieve relatively high cell concentrations in a continuous bioreactor for the conversion of glucose to ethanol.Research sponsored by the Office of Energy Research, U.S. Department of Energy, under contract W-7405-eng-26 with the Union Carbide Corporation.     

Effect of calcium and sodium salts on ethanol production in high sugar fermentation by free cells ofZymomonas mobilis

Summary Addition of calcium carbonate enhanced ethanol production byZymomonas mobilis ZM4 and a mutant (ZMI₂), especially at higher concentrations (200–400 g/L) of glucose and sucrose, as well as at higher temperature (42°C) by the mutant. Calcium and sodium carbonates neutralized the acid produced in the medium and enhanced the ethanol production. The Na salts were less effective in the parent strain and were not favourable for the mutant. Ca²⁺ ions played a direct role in augmenting ethanol production as evidenced by the effect of calcium chloride at controlled pH (5.5).