Bioremediation and decolorization of anaerobically digested distillery spent wash

A facultative anaerobic pure bacterial culture L-2 capable of growth on 12.5% (v/v) diluted digested spent wash supplemented with glucose (10g/l) was isolated from an Indian distillery. It achieved 31% decolorization and 57% COD reduction after 7 days' incubation. The advantages of using such a culture for digested spent wash bioremediation are apparent in providing a realistic approach for decreasing its pollution potential prior to disposal.     

Self-cycling fermentation in a stirred tank reactor

Self-cycling fermentations (SCFs) were conducted in a stirred tank apparatus using Bacillus subtilis and Acinetobacter calcoaceticus. The systems were very stable and the experiments lasted through many cycles. The variation of parameters such as biomass and doubling time from cycle to cycle was small. The stirred tank reactor (STR) allowed a much better control of the working volume in the fermentor from cycle to cycle, compared to the cyclone column, and it was not necessary to make periodic corrections.The production of surfactin from B. subtilis was achieved without extending the cycle time. The harvested broth at the end of each cycle was allowed to remain in a secondary vessel, at ambient temperature, before being collected. It is exhaustion of the limiting nutrient which causes an increase in dissolved oxygen (DO). At this point, the computer, which constantly monitors the DO, triggered the harvesting sequence to end the cycle. Thus, the mature culture in the secondary vessel experienced appropriate conditions for the production of the secondary metabolite. Meanwhile, the next batch of cells was being grown in the primary reactor.The response of a gas analyzer on the effluent paralleled that of the DO measurements in the fermentor. These data for oxygen and carbon dioxide exhibited less noise than the DO readings. Either would be a more reliable parameter for feedback control of the SCF because the problem of fouling of the DO probe after extended runs of many cycles would be eliminated. (c) 1993 John Wiley & Sons, Inc.     

Cephalosporin C production in a stirred tank reactor

Summary Cephalosporin C was produced with the moldCephalosporium acremonium in a 20 1 stirred tank reactor with 100 kg/m³ peanut flour in fed-batch operation. The growth and product formation was followed by on-line analysis of the broth composition. The cell concentration was estimated from the RNA-content of the cells. By optimization of the fed-batch operation and by increasing the phosphate content in the broth, a final cephalosporin C concentration of 12 kg/m³ was attained.Nomenclature CPC cephalosphorin C - DAC deacetylcephalosporin C - DAOC deacetoxycephalosporin C - k _L a volumetric mass transfer coefficient - MMBS 2-Hydroxy-4-methylmercaptobutyric acid - PABAH p-Hydroxybenzoicacidhydrazid - RNA ribonucleic acid - RQ respiratory quotient - oxygen transfer rate - CO₂-production rate - t fermentation time     

A bioprocess for the remediation of anaerobically digested molasses spentwash from biogas plant and simultaneous production of lactic acid

A bacterial isolate LAB-1 identified as Lactobacillus casei was cultivated in a fermentation medium containing biogas plant effluent. This effluent was generated after anaerobic digestion of molasses spentwash in biogas fermenters. In the bioremediation process of this very dark coloured effluent through the cultivation of L. casei, decrease in effluent's colour of 49 and 52% and reduction in COD level up to 54 and 57% were achieved using bacterial cells in free and immobilised system, respectively, in 5 days batch cultivation. During this process of bioremediation, a metabolite lactic acid was produced by this bacterial isolate with the yield of 10.9 and 11.3 mg/ml, in free and immobilised cells fermentation, respectively.     

Biodegradation and decolourization of anaerobically treated distillery spent wash by a novel bacterial consortium

The aim of this study was to isolate microorganisms capable of decolourizing and degrading anaerobically treated distillery spent wash. A bacterial consortium DMC comprising of three bacterial cultures was selected on the basis of rapid effluent decolourization and degradation, which exhibited 67 +/- 2% decolourization within 24 h and 51 +/- 2% chemical oxygen demand reduction within 72 h when incubated at 37 degrees C under static condition in effluent supplemented with 0.5% glucose, 0.1% KH(2)PO(4), 0.05% KCl and 0.05% MgSO(4) x 7H(2)O. Addition of organic or inorganic nitrogen sources did not support decolourization. The cultures were identified as Pseudomonas aeruginosa PAO1, Stenotrophomonas maltophila and Proteus mirabilis by the 16S rDNA analysis.     

Continuous ATP production by spinach thylakoid in a stirred tank reactor

Spinach thylakoid was immobilized by two different methods for the purpose of retention within a continuous-flow stirred tank reactor (CSTR). The glutaraldehyde crosslinked albumin polymer method completely inactivated the cyclic ATP photophosphorylation of thylakoid. In contrast, agarose-entrapped thylakoid retained about 17% of the activity of the cyclic photophosphorylation of non-immobilized thylakoid. This activity declined continuously during ATP production in the CSTR. Fifty percent of the initial activity was lost within about 5.5 h. Ascorbate was found to increase the stability of ATP photophosphorylation; about twice as much ATP was produced at the optimal ascorbate concentration of 5 mM. Under the optimal dilution rate of 2.36 h⁻¹, about 60 μmol of ATP per mg chlorophyll were produced in 20 h by agarose-entrapped thylakoid in the CSTR. These results showed that, compared to non-immobilized thylakoid in batch operation, agarose-entrapped thylakoid produced only a low amount of ATP under continuous operation.     

Study of microencapsulated urease in a continuous feed,stirred tank reactor

Urease, (urea amidohydrolase, EC 3.5.1.5) co-encapsulated with haemoglobin in cellulose nitrate membranes was found to exhibit apparent Michaelis-Menten kinetics; however, a steadily increasing apparent Michaelis-Menten constant over the lifetime of the preparation was observed. The activity of the enzyme in a continuous feed stirred tank reactor (CSTR) was investigated and correlated with a mathematical model derived from basic Michaelis-Menten kinetics. Plots relating substrate conversion to feed substrate concentration and tank reactor capacity were constructed and found to be accurate to less than 15% error under the experimental conditions studied.     

A comparative study of immobilized amyloglucosidase in a packed bed reactor and a continuous feed stirred tank reactor

The catalytic activity of amyloglucosidase covalently attached to DEAE-cellulose was studied in a packed bed reactor and a continuous feed stirred tank reactor (CSTR) for the reaction maltose → glucose. At low flow rates mass-transfer limitations in the bed reactor lead to lower conversions for this reactor compared to the CSTR. Simple theoretical expressions for these reactors were compared with the experimental results. There are significant differences between the kinetic parameters and pH profile of the immobilized and free enzyme. The immobilized enzyme also showed greater stability at 50°C than did free amyloglucosidase. The temperature dependence of the reaction rate was the same for immobilized and free enzyme.     

Optimization of operational conditions for adipate ester synthesis in a stirred tank reactor

Esterification of adipic acid and oleyl alcohol in a solvent-free system featuring a stirred tank reactor containing commercially immobilized Candida antarctica lipase B was performed. The process was carried out using an artificial neural network (ANN) trained by the Levenberg-Marquardt (LM) algorithm. The effects of four operative variables, temperature, time, amount of enzyme, and impeller speed, on the reaction yield were studied. By examining different ANN configurations, the best network was found to consist of seven hidden nodes using a hyperbolic tangent sigmoid transfer function. The values of the coefficient of determination (R²) and root mean squared error (RMSE) between the actual and predicted responses were determined to be 1 and 0.0058178 for training and 0.99467 and 0.622540 for the testing datasets, respectively. These results imply that the developed model was capable of predicting the esterification yield. The operative variables affected the yield, and their order of contribution was as follows: time > amount of enzyme > temperature > impeller speed. A high percentage of yield (95.7%) was obtained using a low level of enzyme (2.5% w/w), and the temperature, time, and impeller speed were 66.5°C, 354 min (about 6 h), and 500 rpm, respectively. A simple protocol for efficient substrate conversion in a solvent-free system evidenced by high enzyme stability is indicative of successful ester synthesis.     

Operational stability of immobilized d-glucose isomerase in a continuous feed stirred tank reactor

d-Glucose isomerization has been studied using immobilized cells of Streptomyces phaeochromogenes in a continuous feed stirred tank reactor (CSTR) where the external film diffusion resistance was negligible. Experiments conducted with various sizes of enzyme particles indicated that a strong internal diffusion resistance improved the apparent stability of these particles. The performance equations of the CSTR were constructed by associating the material balances for the inside porous support matrix with the bulk liquid phase, and enzyme deactivation was also taken into consideration. An iterative method together with the orthogonal collocation method is proposed for the evaluation of effectiveness factor and the substrate concentration profile within the enzyme particles. The numerical results offer an alternative analytical proof for the observation that under strong internal diffusion control the apparent operational stability of immobilized enzyme is improved.     

Bioleaching of chalcopyrite concentrate by a moderately thermophilic culture in a stirred tank reactor

A mixed culture of moderately thermophilic microorganisms was enriched from acid mine drainage samples collected from several chalcopyrite mines in China. Such mixed culture can be used to effectively extract copper from chalcopyrite. Furthermore, after being adapted to gradually increased concentration of chalcopyrite concentrate, the tolerance of the mixed culture to chalcopyrite concentrate was brought up to 80 g/L. The effects of several leaching parameters on copper recovery in stirred tank reactor also had been investigated. The results of the investigation show that it was possible to achieve a copper extraction rate of 75% in 44 days at a pulp density of 8%. The leaching rate of chalcopyrite concentrate tended to increase with dissolved total iron concentration. At low pH ranges, more microscopic counts of microorganisms were found in the solution. Furthermore, the analysis of leached residues indicates that the passivation of chalcopyrite concentrate was mainly due to a mass of jarosite and PbSO(4) on the mineral surface, other than the elemental sulphur layer. The bacterial community composition was analyzed by using Amplified Ribosomal DNA Restriction Analysis. Two moderately thermophilic bacteria species were identified as Leptospirillum ferriphilum and Acidithiobacillus caldus with abundance of 67% and 33% in the bio-pulp, respectively.     

Semi-continuous xylose-to-xylitol bioconversion by Ca-alginate entrapped yeast cells in a stirred tank reactor

Candida guilliermondii FTI 20037 cells were entrapped in Ca-alginate beads and used for xylose-to-xylitol bioconversions during five successive batches in a stirred tank reactor. Supplemented sugarcane bagasse hemicellulosic hydrolysate was used as the fermentation medium. The average volume of the Ca-alginate beads was reduced by about 30% after the 600 h taken to perform the five bioconversion cycles, thus demonstrating physical instability under the conditions prevailing in the reactor vessel. In spite of this, almost steady bioconversion rates and yields were observed along the repeated batches. In average values, a production of 51.6 g l(-1), a productivity of 0.43 g l(-1 )h(-1) and a yield of 0.71 g g(-1) were attained in each batch, variation coefficients being smaller than 10%.     

Influence of medium composition on penicillin V production in a stirred tank reactor.

Penicillin production with a high-producing strain Penicillium chrysogenum was investigated under well-controlled conditions in a stirred tank reactor with complex media containing lard oil and lactose on the one hand, and lactose on the other hand. With lard oil, cell growth and product formation rates were higher, and the production time was shorter by 40 h than without lard oil. On account of the longer production time without lard oil, the amount of beta-lactam compounds was higher (29.93 g l-1), but the mole fraction of the decomposed products (penicilloic acid and penilloic acid) was larger (0.282) than the amount of penicillin V (23.25 g l-1) and the decomposed mole fraction (0.0747) with lard oil. The final product concentrations were about the same (20.86 g l-1 or 35,462 IU ml-1 with lard oil, and 20.43 g l-1 or 34510 IU ml-1 without lard oil). The mole fractions of the by-product (p-OH-penicillin V) were 0.0365 and 0.066. The substitution of lard oil with lactose is possible without a considerable reduction of process performance.     

Disturbance-accommodating control approach applied for the control of a continuous stirred tank reactor

This paper describes an experimental study of linear adaptive control to achieve the monitoring of a continuous stirred tank reactor. The practical control objective was the regulation of the substrate concentration at a pre-specified value in the process effluent despite local changes and/or culture physiology variations. The substrate concentration and the dilution rate have been selected as the controlled and the control variable respectively. The results obtained confirm that this approach offers the possibility to combine simplicity and effectiveness in bioprocess control.     

Hydrogen metabolic patterns driven by Clostridium-Streptococcus community shifts in a continuous stirred tank reactor

Applied Microbiology and Biotechnology - The hydrogen (H2) production efficiency in dark fermentation systems is strongly dependent on the occurrence of metabolic pathways derived from the...     

Optimization of anaerobically digested distillery molasses spent wash decolorization using soil as inoculum in the absence of additional carbon and nitrogen source

The aim of this study was to achieve maximum decolorization of molasses spent wash (MSW) in absence of any additional carbon or nitrogen source using soil as inoculum. Soil samples were collected from the MSW disposal site. Colored soil samples exhibited higher pH, sugar and protein as compare to less colored samples. A decolorization of 69% was obtained using 10% (w/v) soil and 12.5% (v/v) MSW after 7 days incubation. Optimized parameters including days--6 days, pH--6, MSW--12.5% and soil concentration--40%, were obtained for maximum decolorization. A decolorization of 81% was achieved using 10% soil and 12.5% MSW after 18 days incubation in absence of any media supplement. Nearly 12% reduction in decolorization activity of the soil sample was observed over a period of 12 months when stored at 6 degrees C. It could be concluded that the decolorization of MSW might be achieved using soil as inoculum without addition of chemical amendments.     

Co-immobilized aerobic/anaerobic mixed cultures in stirred tank and gaslift loop reactors

Co-immobilized Aspergillus awamori and Zymomonas mobilis cultures were investigated in a stirred tank reactor on synthetic medium with starch as substrate at various dissolved oxygen concentrations. In a gaslift loop reactor, freely suspended and immobilized A. awamori were cultivated on synthetic medium and soluble potato starch. In the same reactor, the growth and ethanol production of freely suspended and immobilized Z. mobilis cultures were studied on synthetic medium and glucose. Co-immobilized A. awamori and Z. mobilis were cultivated in batch and continuous operations in the gaslift loop reactor on synthetic medium with starch substrate at different dissolved oxygen concentrations. The interrelations between the different process variables are discussed.     

Measuring carbon monoxide gas-liquid mass transfer in a stirred tank reactor for syngas fermentation

Carbon monoxide-liquid mass transfer rates in a water-filled stirred tank reactor are determined using a myoglobin protein method to measure dissolved carbon monoxide concentrations as a function of time. Data are acquired over a range of stirrer speeds (200 < or = N < or = 600 rpm) and gas flow rates (1 < or = Q < or = 6 L/min), corresponding to a gas retention time range of 1.2-7 min. Volumetric CO-water mass transfer coefficients range from 0.003 to 0.043 s(-1) and are well-correlated using the power density and the superficial gas velocity.     

Scale-up of biotransformation process in stirred tank reactor using dual impeller bioreactor

The gas-liquid mass transfer coefficient K(L)a in the fermenter is a strong function of mode of energy dissipation and physico-chemical properties of the liquid media. A combination of disc turbine (DT) and pitched blade turbine down flow (PTD) impellers has been tested in laboratory bioreactor for gas hold-up and gas-liquid mass transfer performance for the growth and biotransformation medium for an yeast isolate VS1 capable of biotransforming benzaldehyde to L-phenyl acetyl carbinol (L-PAC) and compared with those in water.Correlations have been developed for the prediction of the fractional gas hold-up and gas-liquid mass transfer coefficient for the above media. The mass transfer coefficient and respiration rate have been determined in the shake flask for the growth as well as for biotransformation medium. These results, then have been used to optimize the operating parameters (impeller speed and aeration) for growth and biotransformation in a laboratory bioreactor. The comparison of cell mass production and L-PAC production in the bioreactor has been done with that obtained in shake flask studies.