Maximum Volumetric Productivity Research Articles

Anaerobic sequencing batch reactor treatment of landfill leachate

Anaerobic treatability of municipal landfill leachate was evaluated using lab-scale anaerobic sequencing batch reactors (ASBR) at 35°C. Experimental studies were conducted at wide-range of volumetric (0.4–9.4 g COD l −1 d −1) and specific (0.2–1.9 g COD g −1 VSS d −1) loading rates by varying hydraulic retention times (HRT) (10–1.5 days) and influent wastewater COD's (3800–15900 mg l −1). The COD removal efficiencies were in the range of 64–85% depending on volumetric and specific loading rates applied. The net daily biomass conversion yields were about 0.1 g volatile suspended solids (VSS) per g of COD removed. Bacterial decay rate constant was 0.01 d −1. The maximum volumetric methane production rate (VMPR) of 1.85 l CH 4 l −1 d −1 was achieved at the volumetric loading rate of 9.4 g COD l −1 d −1. The specific methane production rates in terms of COD were about 1.06 g CH 4–COD g VSS −1 d −1 in the reactor. The results have shown that about 83% of COD removed during the treatment was converted to methane. The average biomass yield was determined as 0.12 g VSS per gram of COD removed.

Modeling and scale-up of the unsterile scleroglucan production process with Sclerotium rolfsii ATCC 15205

An empirical model was applied to describe the growth related formation of scleroglucan in batchwise cultivation of Sclerotium rolfsii. In this case, the level of oxygen supply controls the carbon flux into glucan, biomass, and CO2 evolution and therefore determines the yield coefficients YGlucan/BDM and YBDM/O2. It was observed that scleroglucan formation is enhanced under microaerobic conditions. However, as the empirical model and data of actual batch cultivations show, different maxima exist for product end concentration [g/l] and volumetric productivity [g/ld] depending on the total oxygen uptake during cultivation. A sufficient bulk mixing of the highly viscous culture suspension becomes particularly important during large-scale cultivations. In addition, the scleroglucan production process proved to be shear sensitive. A correlation between the attainable molecular weight of the glucan and the stirrer tip velocity in bioreactors of different sizes is presented. For all these reasons, a scale-up of this process is very complex. Large-scale cultivations under microaerobic conditions, aiming for maximum product end concentration, were slowed down by poor bulk mixing leading to a lower carbon flux into glucan formation. On the other hand, a scale-up designed for maximum volumetric productivity using high oxygen supply was successfully conducted up to a reactor volume of 1.500 l. To minimize the loss in product quality (molecular weight of the glucan) due to high stirrer tip velocities, a mixing concept was developed employing reduced agitation combined with maximum aeration to secure a sufficient axial bulk mixing in the reactor.

Increase of xylitol yield by feeding xylose and glucose in Candida tropicalis.

Candida tropicalis, a strain isolated from the sludge of a factory manufacturing xylose, produced a high xylitol concentration of 131 g/l from 150 g/l xylose at 45 h in a flask. Above 150 g/l xylose, however, volumetric xylitol production rates decreased because of a lag period in cell growth. In fed-batch culture, the volumetric production rate and xylitol yield from xylose varied substantially with the controlled xylose concentration and were maximum at a controlled xylose concentration of 60 g/l. To increase the xylitol yield from xylose, feeding experiments using different ratios of xylose and glucose were carried out in a fermentor. The maximum xylitol yield from 300 g/l xylose was 91% at a glucose/xylose feeding ratio of 15%, while the maximum volumetric production rate of xylitol was 3.98 g l-1 h-1 at a glucose/xylose feeding ratio of 20%. Xylitol production was found to decrease markedly as its concentration rose above 250 g/l. In order to accumulate xylitol to 250 g/l, 270 g/l xylose was added in total, at a glucose/xylose feeding ratio of 15%. Under these conditions, a final xylitol production of 251 g/l, which corresponded to a yield of 93%, was obtained from 270 g/l xylose in 55 h.

Optimization of Phaffia rhodozyma continuous culture through response surface methodology

Response surface methodology was applied to optimize the growth of the yeast Phaffia rhodozyma in continuous fermentation using peat hydrolysates as substrate. A second-order, complete, factorial design of the experiments was used to develop empirical models providing a quantitative interpretation of the relationships between the two variables studied, dilution rate and pH. Maximum biomass concentration in the fermentor was obtained by employing the following predicted optimum fermentation conditions: a dilution rate of 0.017/h and a pH level of 7.19. A verification experiment, conducted at previously optimized conditions for maximum biomass volumetric productivity (a dilution rate of 0.022/h, and a pH level of 6.90), produced values for biomass concentration, residual substrate concentration, biomass yield, and biomass volumetric productivity that were very close to the predicted values, indicating the reliability of the empirical model. The concentration of the pigment astaxanthin produced by the yeast under the optimized growth conditions was found to be 544 mg astaxanthin/kg dry cell biomass. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 314–320, 1998.

Optimization ofPhaffia rhodozyma continuous culture through response surface methodology

Response surface methodology was applied to optimize the growth of the yeast Phaffia rhodozyma in continuous fermentation using peat hydrolysates as substrate. A second-order, complete, factorial design of the experiments was used to develop empirical models providing a quantitative interpretation of the relationships between the two variables studied, dilution rate and pH. Maximum biomass concentration in the fermentor was obtained by employing the following predicted optimum fermentation conditions: a dilution rate of 0.017/h and a pH level of 7.19. A verification experiment, conducted at previously optimized conditions for maximum biomass volumetric productivity (a dilution rate of 0.022/h, and a pH level of 6.90), produced values for biomass concentration, residual substrate concentration, biomass yield, and biomass volumetric productivity that were very close to the predicted values, indicating the reliability of the empirical model. The concentration of the pigment astaxanthin produced by the yeast under the optimized growth conditions was found to be 544 mg astaxanthin/kg dry cell biomass.

γ-ray induced mutagenesis ofCellulomonas biazotea for improved production of cellulases

A rifampin-resistant mutant ofCellulomonas biazotea secreted elevated levels of cellulasesin vivo. The cellulase production in the mutant was not inhibited in the presence of 5% glucose, cellobiose or glycerol in the solid medium. The mutant exhibited approximately two- to three-fold enhanced product yields and productivity of cellular β-glucosidase over the wild parent in shake-flask culture studies when grown on either cellulosic or lignocellulosic substrates. Extracellular production of filter paper cellulase (FPase) and endo-glucanase (CMCase) were also significantly (p≤0.05) altered. During growth of the mutant on α-cellulose, the maximum volumetric productivities for CMCase, FPase and β-glucosidase were 52, 23.3, and 15.2 IUL−1 h−1,i.e 118, 121, and 229% their respective values for the parental strain. Some enzyme properties of the mutant cellulases were altered. Mutant-derived cellulases produced higher yields of glucose arising by degradation of bagasse, wheat straw, and α-cellulose (1.53-, 1.57-, and 1.75-fold, respectively).

Mutagenesis of Cellulomonas biazotea for enhanced production of xylanases

A rifampin-resistant mutant of Cellulomonas biazotea, designated 7 Rf, secreted elevated levels of xylanases in vivo. The xylanase and β-xylosidase production in the mutant was not inhibited in the presence of 5% glucose, cellobiose or glycerol in the solid medium. This mutant showed approximately 2–3-fold enhanced product yield and productivity of cellular β-xylosidase over the wild type parent in shake flask culture studies when grown on either cellobiose, cellulosic or lignocellulosic (LC) substrates. Extracellular production of xylanase was also significantly ( P ≤ 0.05) altered in the mutant. During growth of C. biazotea mutant derivative on xylan, maximum volumetric productivities ( Q p) for xylanase and β-xylosidase were 493, and 30.7 IU/l/h, which were 1.21- and 2.29-fold improved over their respective values from the parental strain. The above values are statistically ( P ≤ 0.05) higher than the values reported in some other xylanolytic mutant organisms.

Biodesulfurization of flue gases and other sulfate/sulfite waste streams using immobilized mixed sulfate-reducing bacteria.

Sulfur dioxide (SO2) is one of the major pollutants in the atmosphere that cause acid rain. Microbial processes for reducing SO2 to hydrogen sulfide (H2S) have previously been demonstrated by utilizing mixed cultures of sulfate-reducing bacteria (SRB) with municipal sewage digest as the carbon and energy source. To maximize the productivity of the bioreactor for SO2 reduction in this study, various immobilized cell bioreactors were investigated: a stirred tank with SRB flocs and columnar reactors with cells immobilized in either potassium-carrageenan gel matrix or polymeric porous BIO-SEP beads. The maximum volumetric productivity for SO2 reduction in the continuous stirred-tank reactor (CSTR) with SRB flocs was 2.1 mmol of SO2/(h.L). The potassium-carrageenan gell matrix used for cell immobilization was not durable at feed sulfite concentrations greater than 2000 mg/L (1.7 mmol/(h.L)). A columnar reactor with mixed SRB cells that had been allowed to grow into highly stable BIO-SEP polymeric beads exhibited the highest sulfite conversion rates, in the range 16.5 mmol/(h.L) (with 100% conversion) to 20 mmol/(h.L) (with 95% conversion). The average specific activity for sulfite reduction in the column, in terms of dry weight of SRB biomass, was 9.5 mmol of sulfite/(h.g). In addition to flue gas desulfurization, potential applications of this microbial process include the treatment of sulfate/sulfite-laden wastewater from the pulp and paper, petroleum, mining, and chemical industries.

Batch and Continuous Production of Lactic Acid from Beet Molasses by Lactobacillus delbrueckii IFO 3202

Process variables were optimized for the production of lactic acid from pretreated beet molasses by Lactobacillus delbrueckii IFO 3202 for batch and continuous fermentations. In the batch fermentation, maximum yields (95·4% conversion, 77·1% effective) and maximum lactic acid volumetric productivity (4·83 g dm−3 h−1) was achieved at 45°C, pH 6·0, 78·2 g dm−3 sugar concentration with 10 g dm−3 yeast extract. Various cheaper nitrogen sources were replaced with yeast extract on equal nitrogen bases in batch fermentation. Of all the nitrogen sources tested, yeast extract yielded the highest and malt sprouts yielded the second highest level of lactic acid. In the continuous fermentation, maximum lactic acid (4·15%) was obtained at a dilution rate of 0·1 h−1. Maximum volumetric lactic acid productivity (11·20 g dm−3 h−1) occurred at D = 0·5 h−1 dilution rate. © 1997 SCI.

Batch and Continuous Production of Lactic Acid from Beet Molasses byLactobacillus delbrueckii IFO 3202

Process variables were optimized for the production of lactic acid from pretreated beet molasses by Lactobacillus delbrueckii IFO 3202 for batch and continuous fermentations. In the batch fermentation, maximum yields (95·4% conversion, 77·1% effective) and maximum lactic acid volumetric productivity (4·83 g dm−3 h−1) was achieved at 45°C, pH 6·0, 78·2 g dm−3 sugar concentration with 10 g dm−3 yeast extract. Various cheaper nitrogen sources were replaced with yeast extract on equal nitrogen bases in batch fermentation. Of all the nitrogen sources tested, yeast extract yielded the highest and malt sprouts yielded the second highest level of lactic acid. In the continuous fermentation, maximum lactic acid (4·15%) was obtained at a dilution rate of 0·1 h−1. Maximum volumetric lactic acid productivity (11·20 g dm−3 h−1) occurred at D = 0·5 h−1 dilution rate. © 1997 SCI.

Process optimization for large-scale production of TGF-alpha-PE40 in recombinant Escherichia coli: effect of medium composition and induction timing on protein expression.

The effects of medium composition and induction timing on expression of a chimeric fusion protein TGF-alpha-PE40 (TP-40) in Escherichia coli strain RR1 were examined using a complex medium at several fermentor scales. Two distinctive phases in E. coli catabolism were identified during fermentation based on preferential utilization between protein hydrolysate and glycerol. Maximum specific and volumetric productivities were achieved by inducing the culture when the cells were switching substrate utilization from protein hydrolysate to glycerol. By increasing the yeast extract concentration in the production medium, initiation of the catabolic switch was delayed until high cell mass was achieved. The final titer of TP-40 at the 15-L fermentation scale was doubled from 400 mg L-1 to 850 mg L-1 by increasing the yeast extract concentration from 1% to 4% (w/v) and delaying the time of induction. This fermentation process was rapidly scaled up in 180-L and 800-L fermentors, achieving TP-40 titers of 740 and 950 mg L-1, respectively.

Effect of inoculum level of xylitol production from rice straw hemicellulose hydrolysate by Candida guilliermondii.

The effect of inoculum level on xylitol production by Candida guilliermondii was evaluated in a rice straw hemicellulose hydrolysate. High initial cell density did not show a positive effect in this bioconversion since increasing the initial cell density from 0.67 g L-1 to 2.41 g L-1 decreased both the rate of xylose utilization and xylitol accumulation. The maximum xylitol yield (0.71 g g-1) and volumetric productivity (0.56 g L-1 h-1) were reached with an inoculum level of 0.9 g L-1. These results show that under appropriate inoculum conditions rice straw hemicellulose hydrolysate can be converted into xylitol by the yeast C. guilliermondii with efficiency values as high as 77% of the theoretical maximum.

Production of lactic acid from whey with agar immobilized cells in a continuous packed tubular reactor

Growth and acid formation of lactic acid bacteria were examined under batch operating conditions for various micro-organism strains in order to find the most efficient strain for lactic acid production. In addition, production of lactic acid from deproteinized whey using the efficient strain immobilized in an agar gel and fermented in a continuous fixed bed reactor was investigated. The optimal processing temperature and dilution rate were found to be 40°C and 0.33 h−1. Maximum volumetric productivities of lactic acid from whey were 7.28 and 9.36 gl−1h−1 without and with recycle, respectively.

Kinetics and stability of a fibrous-bed bioreactor for continuous production of lactic acid from unsupplemented acid whey

The kinetics and long-term stability of the fibrous-bed bioreactor were investigated for continuous lactic acid production from unsupplemented acid whey containing ~ 3.7% (w/v) lactose and ~ 0.8% (w/v) lactic acid, using immobilized cells of Lactobacillus helveticus at pH 5.5 and 42 °C. Depending on dilution rate and lactate concentration, reactor volumetric productivities ranged from 2.6 g l −1 h −1 to 7 g l −1 h −1. These productivities were approx. 10-times higher than those obtained in batch cultivations with free cells. The product inhibition followed noncompetitive kinetics, with the inhibition constant, K p, equal to 7.9 g l −1 and the maximum volumetric productivity, q′ max, equal to 20 g l −1 h −1. Lactate yield from lactose was > 95% (w/w). Reactor performance was stable for continuous, long-term operation for both sterile and nonsterile unsupplemented acid whey feeds for the 6-month period studied. Concentrated acid whey was also found to be an appropriate feedstock to this bioreactor. SEM micrographs showed a dense biofilm covering the fibrous reactor packing. Atypical cell morphology was also observed with the immobilized cells. Total cell density, including suspended cells, within the reactor exceeded 60 g 1 −1. The specific cell productivity was lower than for free cells, indicating a relatively low cell efficiency, perhaps due to diffusion limitation.

Effect of dilution rate in continuous production of l-ornithine by an arginine auxotrophic mutant

Continuous production of l-ornithine by an arginine auxotroph of Brevibacterium ketoglutamicum was carried out in a 2- l jar fermentor. The effects of the dilution rate on the fermentation parameters were investigated under l-arginine limited conditions. The maximum l-ornithine yield and volumetric productivity obtained were 0.213 g/g and 0.313 g/ l/h, respectively. The specific l-ornithine production rate increased with the dilution rate, with a maximum value of 0.084 g/g/h at a dilution rate of 0.125 h −1.

Glucose oxidase, catalase and gluconic acid production by immobilized mycelium of Penicillum variabile P16

Conidia of Penicillium variabile P16 producing glucose oxidase were immobilized in different carriers and used in repeated-batch processes. Limited free-cell growth and good mechanical stability of the carriers were obtained with Ca-alginale, agar and polyurethane sponge. During prolonged experiments, the polyurethane sponge appeared to be the best carrier in relation to glucose oxidase and catalase activities. The maximum mean volumetric productivity of gluconate was obtained using agar as immobilizing agent.

Production of acetone-butanol-ethanol by Clostridium acetobutylicum using a spin filter perfusion bioreactor

The performance of a custom-built cell perfusion bioreactor was investigated in the biosynthesis of acetone, butanol and ethanol by Clostridium acetobutylicum. In a continuous flow operating mode with total cell retention, i.e., when only the cell-free spent medium was removed, steady-state concentrations of acetone, butanol, ethanol, butyric acid, acetic acid and residual glucose and increasing biomass concentration were observed up to 49 g l −1 of glucose in the feed stream. The steady-state concentrations of acetone, butanol, ethanol, butyric acid and acetic acid were independent of the dilution rate and a function of the feed glucose concentration. Under these conditions a complete utilization of glucose with a maximum volumetric productivity of 1.14 g l −1 h −1 of total solvents was obtained. Increasing the feed glucose concentration to 58 and 63 g l −1 in total cell retention mode led to sustained undampened oscillations resulting from product inhibition. These oscillations were eliminated when a full harvest stream of cell containing spent medium was withdrawn in addition to the cell-free filtrate stream.

Optimization of the conditions of the fermentation of beet molasses to lactic acid by Lactobacillus delbrueckii

AbstractOptimization studies were carried out for the production of L‐lactic acid from the fermentation of beet molasses by Lactobacillus delbrueckii. A Central Composite Design was used to determine the optimum values of the process variables (temperature, pH, inoculum concentration, and initial sucrose concentration) for obtaining the maximum yield and the maximum volumetric productivity of lactic acid. Among the variables selected for study, it was found that all of them apart from the temperature significantly affected the responses (yield and volumetric productivity of lactic acid). The Central Composite Design also permitted formulating two second‐order polynomial empirical models relating to the responses and the significant variables. From these models it was possible to determine the value of the variables giving the maximum yield of lactic acid production (87.8%) and the maximum volumetric rate of lactic acid biosynthesis (2.7 g/l · h). Finally, the dependence of the lactic acid yield and productivity on the model variables was investigated. All conclusions are restricted to the experimental range studied.

Immobilization of the slime mould Dictyostelium discoideum for the continuous production of recombinant human antithrombin III

Recombinant vegetative Dictyostelium discoideum cells were immobilized inside a porous matrix by an inorganic membrane that was permeable to nutrients but not to cells, in order to produce recombinant human antithrombin III. Cells so entrapped could reach up to 15 times higher biomass densities compared with organisms growing freely in suspension. The high cell concentration maintained in the immobilized cell bioreactor caused an increase in specific and volumetric productivity. In continuous operation a maximum volumetric antithrombin productivity of 56 ng h-1 ml-1 catalyst bulk volume was attained at a dilution rate of 0.016 h-1. In addition, the good retention of metabolic activity for several weeks as well as the convenient form of storage and regeneration of the catalytic system were shown.

Effects of pH and acetic acid on glucose and xylose metabolism by a genetically engineered ethanologenic Escherichia coli.

Efficient utilization of the pentosan fraction of hemicellulose from lignocellulosic feedstocks offers an opportunity to increase the yield and to reduce the cost of producing fuel ethanol. The patented, genetically engineered, ethanologen Escherichia coli B (pLOI297) exhibits high-performance characteristics with respect to both yield and productivity in xylose-rich lab media. In addition to producing monomer sugar residues, thermochemical processing of biomass is known to produce substances that are inhibitory to both yeast and bacteria. During prehydrolysis, acetic acid is formed as a consequence of the deacetylation of the acetylated pentosan. Our investigations have shown that the acetic acid content of hemicellulose hydrolysates from a variety of biomass/waste materials was in the range 2-10 g/L (33-166 mM). Increasing the reducing sugar concentration by evaporation did not alter the acetic acid concentration. Acetic acid toxicity is pH dependent. By virtue of its ability to traverse the cell membrane freely, the undissociated (protonated) form of acetic acid (HAc) acts as a membrane protonophore and causes its inhibitory effect by bringing about the acidification of the cytoplasm. With recombinant E. coli B, the pH range for optimal growth with glucose and xylose was 6.4-6.8. With glucose, the pH optimum for ethanol yield and volumetric productivity was 6.5, and for xylose it was 6.0 and 6.5, respectively. However, the decrease in growth and fermentation efficiency at pH 7 is not significant. At pH 7, only 0.56% of acetic acid is undissociated, and at 10 g/L, neither the ethanol yield nor the maximum volumetric productivity, with glucose or xylose, is significantly decreased. The "uncoupling" effect of HAc is more pronounced with xylose and the potency of HAc is potentiated in a minimal salts medium. Controlling the pH at 7 provided an effective means of circumventing acetic acid toxicity without significant loss in fermentation performance of the recombinant biocatalyst.