Degradation and decolorization of monosodium glutamate wastewater with <Emphasis Type="Italic">Coriolus versicolor</Emphasis>

Degradation and decolorization of monosodium glutamate wastewater (MSGW) with Coriolus versicolor were firstly carried out. The effects of various operation parameters namely wastewater concentrations, pH, culture time and incidence of sterilization on maximum percentage of degradation and decolorization of wastewater were investigated. Studies of mycelium and enzyme for C. versicolor degradation and decolorization were estimated in this study. Ten percentage of wastewater concentration and pH = 5.0 were found to be the most suitable ones among the other experiments. The highest degradation and decolorization efficiency of wastewater was obtained at the fifth day of cultivation, which was displayed with more than 70% chemical oxygen demand removal, 83% total sugar removal and 55% color removal, respectively. Sterile operation had no remarkable effect on the degradation and decolorization efficiency for C. versicolor. Mycelium and the extra cellular fungal enzyme were both necessary for the degradation and decolorization of MSGW. C. versicolor possesses great potential and economic advantages in MSGW treatment.     

Purification and characterization of a novel alkaline protease from Bacillus horikoshii

An investigation was conducted on the enhancement of production and purification of an oxidant and SDS-stable alkaline protease (BHAP) secreted by an alkalophilic Bacillus horikoshii, which was screened from the body fluid of a unique Korean polychaeta (Periserrula leucophryna) living in the tidal mud flats of Kwangwha Island in the Korean West Sea. A prominent effect on BHAP production was obtained by adding 2% maltose, 1% sodium citrate, 0.8% NaCl, and 0.6% sodium carbonate to the culturing medium. The optimal medium for BHAP production contained (g/l) SBM, 15; casein, 10; K(2)HPO(4), 2; KH(2)PO(4), 2; maltose, 20; sodium citrate, 10; MgSO(4), 0.06; NaCl, 8; and Na(2)CO(3), 6. A protease yield of approximately 56,000 U/ml was achieved using the optimized medium, which is an increase of approximately 5.5-fold compared with the previous optimization (10,050 U/ml). The BHAP was homogenously purified 34-fold with an overall recovery of 34% and a specific activity of 223,090 U/mg protein using adsorption with Diaion HPA75, hydrophobic interaction chromatography (HIC) on Phenyl-Sepharose, and ion-exchange chromatography on a DEAE- and CMSepharose column. The purified BHAP was determined a homogeneous by SDS-PAGE, with an apparent molecular mass of 28 kDa, and it showed extreme stability towards organic solvents, SDS, and oxidizing agents. The K(m) and k(cat) values were 78.7 μM and 217.4 s(-1) for N-succinyl-Ala- Ala-Pro-Phe-pNA at 37° C and pH 9, respectively. The inhibition profile exhibited by PMSF suggested that the protease from B. horikoshii belongs to the family of serine proteases. The BHAP, which showed high stability against SDS and H(2)O(2), has significance for industrial application, such as additives in detergent and feed industries.     

Decolorization of molasses wastewater by Lactobacillus plantarum No. PV71-1861

Lactobacillus plantarum No. PV71-1861, isolated from pickle samples in Thailand, showed the high potential for use in decolorization of molasses wastewater under both anaerobic and facultative (static) conditions. The strain showed the highest melanoidin pigment (MP) decolorization yield of 68.12% with MP solution (color intensity corresponding to an optical density of 3.5 units at 475 nm) containing 2% glucose, 0.4% yeast extract, 0.1% KH(2)PO(4), 0.05% MgSO(4).7H(2)O and initial pH of 6 under static condition at 30 degrees C within 7 days. But, it showed low growth and MP decolorization yields under aerobic conditions. Gel filtration chromatograms of the MP solutions showed that the small molecular weight fraction of MP solution was decolorized by the strain when the large molecular weight fraction still remained in the effluent. For application, the strain could apply to treat anaerobic treated-molasses wastewater (T-MWW) with high removal efficiency. The highest MP removal efficiencies and growth yield of 76.6% and 2.6 mg/mL, respectively, were observed with the T-MWW within 7 days of culture, and the effluent pH of the system was decreased to lower than 4.0 after 2-3 days operation.     

Improvement of the decolorization of azo dye by anaerobic sludge bioaugmented with<Emphasis Type="Italic">Desulfovibrio desulfuricans</Emphasis>

A culture of anaerobic sludge was bioaugmented withDesulfovibrio desulfuricans for the color removal of authentic textile wastewater containing a substantial amount of sulfate, in order to improve the decolorization process. The sulfide produced by sulfate respiration ofD. desulfuricans can chemically reduce azo bonds to produce a colorless metabolite in the form of aromatic amines. In the case where the culture of anaerobic sludge was bioaugmented withD. desulfuricans, the decolorization of C.I. Reactive Black 5 showed an increase of more than 14% after 48 h in comparison with that in the culture of anaerobic sludge alone. In the decolorization of authentic textile wastewater, the color removal (about 69.0%) was improved by the mixed culture of anaerobic sludge andD. desulfuricans, compared with results obtained with only anaerobic sludge as reported in our previous work, suggesting that bioaugmentation byD. desulfuricans can be useful for the decolorization of wastewater that contains complex dye compounds and sulfate.     

Decolorization of reactive dyes by mixed cultures isolated from textile effluent under anaerobic conditions

In the present study mixed cultures that could grew in the molasses media were isolated from textile dye effluent and its decolorization activity was studied in a batch system under anaerobic conditions, in order to determine the optimal conditions required for the highest decolorization activity. The optimum pH value for decolorization was determined as 8 for all the dyes tested. In the experiment with pH 8 dye decolorizations by mixed cultures were investigated at about 96.2–1031.3 mg l⁻¹ initial dye concentrations. The highest dye removal rates of mixed cultures were 94.9% for Reactive Red RB, 91.0% for Reactive Black B and 63.6% for Remazol Blue at 953.2, 864.9 and 1031.3 mg l⁻¹ initial dye concentrations respectively within 24 h incubation period. When the Reactive Red RB was used, approximately 82–98% total color removal was obtained at between 96.2 and 953.2 mg l⁻¹ initial dye concentrations after 12 h of incubation at 35 °C. These results show that our enriched mixed cultures have the potential to serve as an excellent biomass for the use in reactive dye removal from wastewaters under anaerobic conditions.     

Biochemical degradation pathway of reactive blue 13 by Candida rugopelliculosa HXL-2

A strain HXL-2 from a lab-scale sequence batch reactor (SBR) was identified as Candida rugopelliculosa based on its physiological, biochemical characteristics, and 26S rDNA D1/D2 gene phylogenetic analysis. About 90% of the 50 mg/L Reactive blue 13(RB13) was degraded in 48 h after inoculation with strain HXL-2. The optimum efficiency of pH on decolorization was obtained at pH 5.The optimum efficiency temperature of C. rugopelliculosa HXL-2 decolorization RB13 was obtained at 28 °C. The color removal efficiency was obtained at 80.3% when the feed concentration reached 2000 mg/L. We first detected naphthalene-like compound is produced as degradation intermediate after the cleavage of RB13 azo bond, detected 1-chloro-3-aniline- 2,4,6-triazine. We proposed degradation pathway of Reactive blue 13 by Candida sp and proved RB13 degradation pathway by Candida sp. has some difference from RB13 degradation pathway by Pseudomonas sp.     

Decolorization of the textile dyes by newly isolated bacterial strains

Six bacterial strains with the capability of degrading textile dyes were isolated from sludge samples and mud lakes. Aeromonas hydrophila was selected and identified because it exhibited the greatest color removal from various dyes. Although A. hydrophila displayed good growth in aerobic or agitation culture (AGI culture), color removal was the best in anoxic or anaerobic culture (ANA culture). For color removal, the most suitable pH and temperature were pH 5.5-10.0 and 20-35 degrees C under anoxic culture (ANO culture). More than 90% of RED RBN was reduced in color within 8 days at a dye concentration of 3,000 mg l(-1). This strain could also decolorize the media containing a mixture of dyes within 2 days of incubation. Nitrogen sources such as yeast extract or peptone could enhance strongly the decolorization efficiency. In contrast to a nitrogen source, glucose inhibited decolorization activity because the consumed glucose was converted to organic acids that might decrease the pH of the culture medium, thus inhibiting the cell growth and decolorization activity. Decolorization appeared to proceed primarily by biological degradation.     

Decolorization of azo dye using PVA-immobilized microorganisms

A microbial consortium having a high capacity for rapid decolorization of azo dye (RED RBN) was immobilized by a phosphorylated polyvinyl alcohol (PVA) gel. The immobilized-cell beads exhibited a color removal capability of 75%, even at a high concentration of RED RBN (500 mg l(-1)) within 12 h using flask culture. The continuous operation was conducted at a hydraulic retention time (HRT) of 5-20 h in which the dye loading rate ranged from 240 to 60 mg dye h(-1). A removal efficiency exceeding 90% was obtained at the HRT higher than 10 h. No recognizable destruction of bead appearance was observed in the 6-month operation. Examination of the mechanism of the decolorization process by cell beads indicated that it proceeded primarily by biological decolorization associated with partial adsorption of the dye onto the entrapped cells and gel matrix. Microscopic observation revealed that the microbial consortium contained in the gel beads was at least made up of three kinds of bacterial species. From the economical viewpoint, alternative cheaper nitrogen sources such as fish meal, soybean meal, pharmamedia and vita yeast powder were examined.     

Decolorization of anthraquinone dye by Shewanella decolorationis S12

A new species of genus Shewanella, Shewanella decolorationis S12, from activated sludge of a textile-printing wastewater treatment plant, can decolorize Reactive Brilliant Blue K-GR, one kind of anthraquinone dye, with flocculation first. Although S. decolorationis displayed good growth in an aerobic condition, color removal was the best in an anaerobic condition. For color removal, the most suitable pH values and temperatures were pH 6.0–8.0 and 30–37°C under anaerobic culture. More than 99% of Reactive Brilliant Blue K-GR was removed in color within 15 h at a dye concentration of 50 mg/l. Lactate was the suitable carbon source for the dye decolorization. A metal compound, HgCl₂, had the inhibitory effect on decolorization of Reactive Brilliant Blue K-GR, but a nearly complete decolorization also could be observed at a HgCl₂ concentration of 10 mg/l. The enzyme activities, which mediate the tested dye decolorization, were not significantly affected by preadaptation of the bacterium to the dye.     

Assembly of mutations for improving thermostability of <Emphasis Type="Italic">Escherichia coli</Emphasis> AppA2 phytase

Conventional processes for lignocellulose-to-organic acid conversion requires pretreatment, enzymatic hydrolysis, and microbial fermentation. In this study, lime-treated wheat straw was hydrolyzed and fermented simultaneously to lactic acid by an enzyme preparation and Bacillus coagulans DSM 2314. Decrease in pH because of lactic acid formation was partially adjusted by automatic addition of the alkaline substrate. After 55 h of incubation, the polymeric glucan, xylan, and arabinan present in the lime-treated straw were hydrolyzed for 55%, 75%, and 80%, respectively. Lactic acid (40.7 g/l) indicated a fermentation efficiency of 81% and a chiral l(+)-lactic acid purity of 97.2%. In total, 711 g lactic acid was produced out of 2,706 g lime-treated straw, representing 43% of the overall theoretical maximum yield. Approximately half of the lactic acid produced was neutralized by fed-batch feeding of lime-treated straw, whereas the remaining half was neutralized during the batch phase with a Ca(OH)₂ suspension. Of the lime added during the pretreatment of straw, 61% was used for the neutralization of lactic acid. This is the first demonstration of a process having a combined alkaline pretreatment of lignocellulosic biomass and pH control in fermentation resulting in a significant saving of lime consumption and avoiding the necessity to recycle lime.     

Investigation of the decolorization efficiency of two pin-to-plate corona discharge plasma system for industrial wastewater treatment

In this article, a dual pin-to-plate high-voltage corona discharge system is introduced to study experimentally the gap distance, the contact time, the effect of pin and plate materials, the thickness of ground plate and the conductivity on the amount of Acid Blue 25 dye color removal efficiency from polluted water. A study for the optimum air gap distance between dual pin and surface of Acid Blue 25 dye solution is carried out using 3D-EM simulator to find maximum electric field intensity at the tip of both pins. The outcomes display that the best gap for corona discharge is approximately 5 mm for 15-kV source. This separation is constant during the study of other factors. In addition, an investigation of the essential reactive species responsible for oxidation of the dye organic compounds (O₃ in air discharge, O₃ in water, and H₂O₂) during the experimental time is conducted. Three various materials such as: stainless steel, copper and aluminum are used for pins and plate. The maximum color removal efficiencies of Acid Blue 25 dyes are 99.03, 82.04, and 90.78% after treatment time 15 min for stainless steel, copper, and aluminum, respectively. Measurement results for the impact of thickness of an aluminum ground plate on color removal competence show color removal efficiencies of 86.3, 90.78, and 98.06% after treatment time 15 min for thicknesses of 2, 0.5, and 0.1 mm, respectively. The increasing of the solution conductivity leads to the reduction of decolorization efficiency. A kinetic model is used to define the performance of corona discharge system. The models of pseudo-zero-order, pseudo-first-order, and pseudo-second-order reaction kinetics are utilized to investigate the decolorization of Acid Blue 25 dye. The rate of degradation of Acid Blue 25 dye follows the pseudo-first-order kinetics in the dye concentration.     

Repeated batch process for biological treatment of black liquor using brown-rot basidiomycete <Emphasis Type="Italic">Fomitopsis</Emphasis> sp. IMER2

A repeated batch operation is developed for the treatment of alkaline pulp black liquor, through a process of biological acidification precipitation of lignin using brown rot fungus Fomitopsis sp. IMER2. The results showed that COD and color removal of black liquor was dependent on the biomass concentration, pH decrease and initial COD. Based on these results, the repeated batch process was successfully carried out 12 times over 36 days in an air bubble column bioreactor. The average reduction of COD and color was approximately 40% and 70%, respectively.     

Purification and partial characterization of serine protease from thermostable alkalophilic <Emphasis Type="Italic">Bacillus laterosporus</Emphasis>-AK1

An extracellular thermostable alkaline protease isolated from Bacillus laterosporus-AK1 was purified by sephadex G-200 gel filtration and DEAE cellulose ion-exchange chromatography techniques. The purified protease showed a maximum relative activity of 100% on casein substrate and appeared as a single band on SDS-PAGE with the molecular mass of 86.29 kDa. The protease was purified to 11.1-folds with a yield of 34.3%. Gelatin zymogram also revealed a clear hydrolytic zone due to proteolytic activity, which corresponded to the band obtained with SDS-PAGE. The protease enzyme had on optimum pH of 9.0 and exhibited highest activity at 75°C. The enzyme activity was highly susceptible to the specific serine protease inhibitor PMSF, suggesting the presence of serine residues at the active sites. Enzyme activity strongly enhanced by the metal ions Ca²⁺ and Mg²⁺ and this enzyme compatible with aril detergent stability retained 75% even 1-h incubation. The purified protease remove bloodstain completely when used with Wheel detergent.     

Decolorization of sugar syrups using commercial and sugar beet pulp based activated carbons

Sugar syrup decolorization was studied using two commercial and eight beet pulp based activated carbons. In an attempt to relate decolorizing performances to other characteristics, surface areas, pore volumes, bulk densities and ash contents of the carbons in the powdered form; pH and electrical conductivities of their suspensions and their color adsorption properties from iodine and molasses solution were determined. The color removal capabilities of all carbons were measured at 1/100 (w/w) dosage, and isotherms were determined on better samples. The two commercial activated carbons showed different decolorization efficiencies; which could be related to their physical and chemical properties. The decolorization efficiency of beet pulp carbon prepared at 750 degrees C and activated for 5h using CO2 was much better than the others and close to the better one of the commercial activated carbons used. It is evident that beet pulp is an inexpensive potential precursor for activated carbons for use in sugar refining.     

Alkaline protease production by an actinomycete MA1-1 isolated from marine sediments

Alkaliphilic actinomycetes isolated from sediment samples of the Izmir Gulf, Turkey were studied for the production of protease activity. Strain MA1-1 was selected as a good alkaline protease producer as measured by the clear zone diameter by the hydrolysis of skim-milk and casein. The alkaline protease production from the marine alkaliphilic actinomycete MA1-1 was studied by using different carbon and nitrogen sources in medium containing glycerol, peptone, KCl, MgSO₄, K₂HPO₄, and trace elements at 30°C for 72 h. Among the different carbon and nitrogen sources, fructose, starch, maltose, D(+) glucose, yeast extract, malt extract, beef extract and peptone provided higher production of protease. Starch was also found to be effective for growth and enzyme production with highest specific activity at 699 U mg^?1. Purification was achieved by adsorption on Diaion HP 20 which resulted in a recovery rate of 68% with a specific activity of 7618 U mg^?1 protein and 40-fold purification. The optimum pH and temperature of the partially purified protease were determined as pH 9.0 and 50°C, but high activity was also observed at pH 8.0–13.0 and 35–50°C. The inhibition profile exhibited by phenylmethylsulphonyl fluoride (PMSF) showed that this enzyme belongs to the serine-protease group.     

Electrochemical oxidation for the treatment of textile industry wastewater

This study elucidates the reduction of organics from textile effluents through electrochemical oxidation technique. Effect of pH and current intensity were investigated in this system. It was found that degradation was maximum at the current intensity of 0.6 A and at a pH of 1.3. Under the same experimental conditions the removal of chemical oxygen demand (COD), total solids, total dissolved solids and total organic carbon were found to be approximately 68%, 49.2%, 50.7% and 96.8%, respectively. Effect of current intensity on color removal was also investigated as a function of electrolysis time (30-210 minutes) and it showed that maximum removal efficiency (96%) was reached within 60 minutes at 0.6 A. While studying the effect of pH on COD removal, it was observed that a decrease in pH to an optimum of 1.3 showed maximum COD reduction of 68%. These results suggest an important role of these parameters in electrochemical process for removing organic pollutants.     

Biosynthesis and properties of an extracellular thermostable serine alkaline protease from <Emphasis Type="Italic">Virgibacillus pantothenticus</Emphasis>

In this communication, we report the presence of a newly identified serine alkaline protease producing bacteria, Virgibacillus pantothenticus (MTCC 6729) in the fresh chicken meat samples and the factors affecting biosynthesis as well as characterization of protease. The strain produced only 14.3 U ml⁻¹ protease in the standard medium after 72 h of incubation, while in optimized culture conditions the production of protease was increased up to 18.2 U ml⁻¹. The strain was able to produce protease at 40°C at pH 9.0. The addition of dextrose and casein improved protease production. The protease was partially purified and characterized in terms of pH and temperature stability, effect of metal ions and inhibitors. The protease was found to be thermostable alkaline by retaining its 100% and 85% stability at pH 10.0 and at 50°C respectively. The protease was compatible with some of the commercial detergents tested, and was effective in removing protein stains from cotton fabrics. The V. pantothenticus, MTCC 6729 protease appears to be potentially useful as an additive in detergents as a stain remover and other bio-formulations.     

Oxidant and SDS-stable alkaline protease from Bacillus clausii I-52: production and some properties

AIMS: An investigation was carried out on an oxidative and SDS-stable alkaline protease secreted by Bacillus clausii of industrial significance. METHODS AND RESULTS: Maximum enzyme activity was produced when the bacterium was grown in the medium containing (g l-1): soyabean meal, 15; wheat flour, 10; liquid maltose, 25; K2HPO4, 4; Na2HPO4, 1; MgSO4.7H2O, 0.1; Na2CO3, 6. The enzyme has an optimum pH of around 11 and optimum temperature of 60 degrees C. The alkaline protease showed extreme stability towards SDS and oxidizing agents, which retained its activity above 75 and 110% on treatment for 72 h with 5% SDS and 10% H2O2, respectively. Inhibition profile exhibited by phenylmethylsulphonyl fluoride suggested that the protease from B. clausii belongs to the family of serine proteases. CONCLUSIONS: Bacillus clausii produced high levels of an extracellular protease having high stability towards SDS and H2O2. SIGNIFICANCE AND IMPACT OF THE STUDY: The alkaline protease from B. clausii I-52 is significant for an industrial perspective because of its ability to function in broad pH and temperature ranges in addition to its tolerance and stability in presence of an anionic surfactant, like SDS and oxidants like peroxides and perborates. The enzymatic properties of the protease also suggest its suitable application as additive in detergent formulations.     

Removal of proteases from <Emphasis Type="Italic">Clostridium perfringens</Emphasis> fermented broth by aqueous two-phase systems (PEG/citrate)

In order to reduce the toxicity of Clostridium perfringens fermentation broths used in vaccine preparation, we developed two-phase aqueous systems for removal of toxin-activating proteases. Removal of the proteases inhibits the conversion of protoxin to active toxin. In order to establish the conditions under which the phase separation occurs, binodal curves, formed by poly(ethylene glycol) (PEG) and sodium citrate, were investigated at different values of pH and PEG molar mass. A 2⁴-experimental design was used to evaluate the influence of PEG molar mass and concentration, citrate concentration and pH on protease partition coefficient, removal factor and protease removal yield. It has been found that simultaneous increase in PEG molar mass and decrease in citrate concentration remarkably improved the removal factor, whereas the protease removal yield showed an opposite trend. The best conditions for the system under consideration (removal factor of 2.69 and yield of 116%) were obtained at pH 8.0 using PEG molar mass of 8000 g mol⁻¹ and concentrations of PEG and citrate of 24 and 15%, respectively.     

Production of fuel ethanol from bamboo by concentrated sulfuric acid hydrolysis followed by continuous ethanol fermentation

An efficient process for the production of fuel ethanol from bamboo that consisted of hydrolysis with concentrated sulfuric acid, removal of color compounds, separation of acid and sugar, hydrolysis of oligosaccharides and subsequent continuous ethanol fermentation was developed. The highest sugar recovery efficiency was 81.6% when concentrated sulfuric acid hydrolysis was carried out under the optimum conditions. Continuous separation of acid from the saccharified liquid after removal of color compounds with activated carbon was conducted using an improved simulated moving bed (ISMB) system, and 98.4% of sugar and 90.5% of acid were recovered. After oligosaccharide hydrolysis and pH adjustment, the unsterilized saccharified liquid was subjected to continuous ethanol fermentation using Saccharomycescerevisiae strain KF-7. The ethanol concentration, the fermentation yield based on glucose and the ethanol productivity were approximately 27.2 g/l, 92.0% and 8.2 g/l/h, respectively. These results suggest that the process is effective for production of fuel ethanol from bamboo.