Decolorization of molasses wastewater by yeast strain, Issatchenkia orientalis No. SF9-246

Among 2402 strains of yeast isolated from various sources in Thailand, a strain No. SF9-246 identified as Issatchenkia orientalis, showed the highest potential for use in decolorization of molasses wastewater. In a malt extract-glucose-peptone broth (MYGP) culture containing melanoidin pigment (MP) at 30 °C a 60.2% decolorization was obtained within 7 days. The strain appeared to enhance both MP-degradation and MP-adsorption. The strain showed MP, chemical oxygen demand (COD) and biological oxygen demand (BOD₅) removal efficiencies of 91.2%, 80.0% and 77.4%, respectively from anaerobic-treated molasses wastewater solution (T-MWW), collected from an anaerobic pond. The wastewater contained 2.5% glucose, 0.1% NH₄Cl, and 0.1% KH₂PO₄. The pH was adjusted to 5.0 at 30 °C for 7 days batch type culture system. The strain showed almost constant decolorization yield of 75–80% over 7 days in a periodical feeding system of 10% fresh T-MWW with the culture system. The strain provided a constant decolorization yield about 70% during 3 replacement cycles. Gel filtration chromatography showed that larger molecular weight fraction of MP solution was rapidly removed, while the smaller molecular weight fraction remained in the effluent.     

Decolorization of Molasses Waste Water by a Thermophilic Strain,Aspergillus fumigatus G-2-6 Sadahiro

We screened for fungi that can decolorize molasses melanoidin in the tropical zone and isolated some strains, mainly in the genus Aspergillus. Of these, strain No. G-2–6 was most active and was identical with Aspergillus fumigatus based on detailed morphological studies.

This strain decolorized about 75% of a molasses melanoidin solution when the strain was cultivated on a glycerol-peptone medium at 45°C for 3 days with shaking. In successive decolorization reusing the mycelia, this strain had more than 60% of the melanoidin-decolorizing activity at the eighth replacement in the presence of 4% glycerol.

Continuous decolorization of molasses melanoidin solution in a jar fermentor had an almost constant decolorization yield of about 70% at a dilution rate of 0.014 hr^-1. At the same time, about 51 % of the chemical oxygen demand and 56% of the total organic carbon in the initial solution were removed. In contrast, continuous decolorization of non-dialyzed molasses melanoidin solution removed a little more chemical oxygen demand and total organic carbon than those of dialyzed molasses melanoidin solution, but had a lower level of melanoidin-decolorizing activity (about 40%).     

Isolation and characterization of distillery spent wash color reducing bacteria and process optimization by Taguchi approach

The ability of five different types of bacterial strains isolated from a distillery mill site was analyzed for decolorization of distillery spent wash. 16S rDNA based denaturing gradient gel electrophoresis (DGGE) and amplified random DNA restriction analysis (ARDRA) were used to characterize the bacterial strains. One of the isolates had higher capability to reduce color (21%) and chemical oxygen demand (COD) (30%) was finally identified by 16S rDNA sequence analysis as Bacillus sp. Different parameters such as pH, temperature, aeration,% carbon,% nitrogen, inoculum size and incubation time were optimized by the Taguchi approach to achieve maximum decolorization of distillery spent wash by the Bacillus sp. Reduction in color (85%) and COD (90%) was observed within 12 h after optimization by the Taguchi method. The significant factor in the optimization process was duration followed by inoculum size to attain maximum color reduction. The Taguchi approach proved to be a reliable tool in optimizing culture conditions and analyzing interaction effects of process parameters in achieving the best possible combination for maximum decolorization of the distillery spent wash.     

Decolorization of anaerobically digested molasses spent wash by <Emphasis Type="Italic">Pseudomonas putida</Emphasis>

The distillery wastewater (spent wash) contains dark brown colored recalcitrant organic compounds that are not amenable to conventional biological treatment. The characteristic recalcitrance to decolorization is due to the presence of brown melanoidin polymers. In the present study, feasibility of using Pseudomonas putida putida strain U for decolorization of spent wash was demonstrated. Batch cultures of P. putida decolourized spent wash by 24%, twofold higher decolorization was achieved following immobilization in calcium alginate beads. Glucose concentration was critical for decolourization, and improved color removal efficiency was obtained by periodic replenishment of glucose. Decolourization was also observed with lactose or whey as alternative carbon sources. The results of our study suggest that P.putida could be used for biological decolorization of molasses spent washes and that supplementation with whey (a by-product from cheese industry) can offer economical viability to the process. Published in Russian in Prikladnaya Biokhimiya i Mikrobiologiya, 2009, Vol. 45, No. 1, pp. 78–83. The text was submitted by the authors in English.     

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.     

Decolorization and biotransformation of triphenylmethane dye, methyl violet, by Aspergillus sp. isolated from Ladakh, India

Methyl violet, used extensively in the commercial textile industry and as a biological stain, is a hazardous recalcitrant. Aspergillus sp. strain CB-TKL-1 isolated from a water sample from Tsumoriri Lake, Karzok, Ladakh, India, was found to completely decolorize methyl violet within 24 h when cultured under aerobic conditions at 25 degrees C. The rate of decolorization was determined by monitoring the decrease in the absorbance maxima of the dye by UV-visible spectroscopy. The decolorization of methyl violet was optimal at pH 5.5 and 30 degrees C when agitated at 200 rpm. Addition of glucose or arabinose (2%) as a carbon source and sodium nitrate or soyapeptone (0.2%) as a nitrogen source enhanced the decolorization ability of the culture. Furthermore, the culture exhibited a maximum decolorization rate of methyl violet after 24 h when the C:N ratio was 10. Nine N-demethylated decolorized products of methyl violet were identified based on UV-visible spectroscopy, Fourier transform infrared (FTIR), and LC-MS analyses. The decolorization of methyl violet at the end of 24 h generated mono-, di-, tri-, tetra-, penta-, and hexa-Ndemethylated intermediates of pararosaniline. The variation of the relative absorption peaks in the decolorized sample indicated a linear decrease of hexa-N-demethylated compounds to non-N-demethylated pararosaniline, indicating a stepwise N-demethylation in the decolorization process.     

Screening of Microorganisms to Decolorize a Model Melanoidin and the Chemical Properties of a Microbially Treated Melanoidin

A microbe of Streptomyces werraensis TT 14, which was newly isolated from soil, decolorized the model melanoidin prepared from glucose and glycine, the decolorization rate being 64% in the optimal medium of pH 5.5 (2.0% starch, 1.0% yeast, 0.3% NaCl, and 0.3% CaC0₃) and 45% in a synthetic medium. There was virtually no difference in the UV-VIS absorption spectra of the microbially treated melanoidin and the control. The peaks of the gel permeation chromatogram for the treated melanoidin and for the control showed the same retention times, but lower molecular weight compounds increased in the decolorized melanoidin. The Cu(II)-chelating activity of the decolorized melanoidin was reduced to about half that of the control. The melanoidin component of pI 2.5 was increased and that of pI 3.5 was reduced by the microbial treatment.     

Production of Decolorizing Activity for Molasses Pigment by Coriolus versicolor Ps4a

The distribution of molasses pigment (melanoidin) decolorizing activity (MDA) was investigated in various Basidiomycetes. MDA was only found in some genera of the white-rot-fungi group of which Coriolus versicolor Ps4a showed high activity, a decolorization yield of approximately 80% under the optimal conditions. Production of MDA by C. versicolor was almost completely coincident with the growth of mycelia. The main MDA was due to intracellular enzymes and induced by the molasses pigment. The induced enzyme consisted of two types, namely a sugar dependent enzyme and a sugar independent enzyme. The decolorization by C. versicolor was due to the decomposition of the molasses pigment.     

Bioremediation Perspective of Navy Blue Rx–Containing Textile Effluent by Bacterial Isolate

The aim of the present study was to investigate the textile effluent degrading potential of an isolated bacterium, Proteus sp. SUK7. The strain had the capacity to decolorize Navy Blue Rx–containing textile effluent up to 83% within 96 h. The maximum decolorization was observed under static conditions at pH 7.0 and 30°C. Reduction in the chemical oxygen demand (COD) and biological oxygen demand (BOD) of textile effluent was observed after treatment with Proteus sp. SUK7. Induction in the activities of laccase and aminopyrine N-demethylase was observed after decolorization, which indicates involvement of these enzymes in the decolorization process. The presence of various inducers was also found to have a modulatory effect on enzyme activities and the decolorization process. Biodegradation was confirmed using various analytical techniques, such as ultraviolet-visible (UV-Vis) spectroscopy, Fourier transform infrared (FTIR), gas chromatography–mass spectrometry (GC-MS), and high-performance liquid chromatography (HPLC). A phytotoxicity study was performed to confirm the nontoxic nature of the degradation metabolites.     

Distillery and curd whey wastes as viable alternative sources for biosurfactant production

Biosurfactant production from synthetic medium and industrial waste, viz. distillery and whey wastes was investigated by using an oily sludge isolate Pseudomonas aeruginosa strain BS2. In synthetic medium separately supplemented with glucose and hexadecane as water-soluble and -insoluble carbon sources, respectively, strain BS2 reduced the surface tension of the fermentation broth from 57 to 27 mN/m. The culture produced biosurfactant during the stationary growth phase and its yield was 0.97 g/l. The culture utilized distillery and whey wastes for its growth, as maximum cell counts reached to 54 × 10⁸ and 64 × 10⁹ c.f.u./ml from an initial inoculum size of 1 × 0⁵ c.f.u./ml, respectively, within 48 h of incubation and in these wastes the yields of biosurfactant obtained were 0.91 and 0.92 g/l, respectively. In synthetic medium, distillery and whey wastes, strain BS2 produced a crystalline biosurfactant which belonged to the category of secondary metabolites and its maximum production occurred after the onset of nitrogen-limiting conditions. After recovering biosurfactant from the fermented waste, the chemical oxygen demand (COD) of distillery and whey wastes was significantly reduced by 81 and 87%, respectively. Total acids, nitrogen and phosphate levels in distillery waste were reduced by 90, 92 and 92%, respectively, while in case of whey waste the concentration of these nutrients was reduced by 88, 95 and 93%, respectively. The isolated biosurfactant possessed potent surface active properties, as it effectively reduced the surface tension of water from 72 to 27 mN/m and formed 100% stable emulsions of a variety of water-insoluble compounds such as hydrocarbons, viz. hexadecane, crude oil, kerosene and oily sludge and pesticides, viz. dichlorodiphenyltrichloroethane (DDT) and benzene hexachloride (BHC). The effectiveness of biosurfactant was also evident from its low critical micellar concentration (CMC) which was 0.028 mg/ml.     

Sucrose-supplemented distillery spent wash as a medium for production of ethanol at 45 °C by free and alginate-immobilized preparations of Kluyveromyces marxianus IMB3

Ethanol production by the thermotolerant, ethanol-producing yeast strain Kluyveromyces marxianus IMB3, was compared during growth on sucrose-supplemented laboratory-based media and distillery spent wash from the Old Bushmill's Distillery Co., Ltd., Co. Antrim, Northern Ireland. Fermentations were carried out using preparations of the free and alginate-immobilized microorganism as inocula in media supplemented with 2 and 10% (w/v) sucrose. Maximum ethanol concentrations accounted for 75–99% of the maximum theoretical yield and in all cases maximum concentrations obtained using the spent wash were similar if not slightly higher than those obtained on the sucrose-supplemented yeast growth media. In addition, the highest concentrations of ethanol were produced by the alginate-immobilized biocatalyst on both types of media. Analysis of exhausted media in the spent wash-based systems demonstrated significant decreases in the total organic carbon content following fermentation. These results confirm our earlier suggestion that ethanol production based on this microorganism in a recycle system may provide a more cost-effective means of disposing of whiskey distillery spent wash.     

Ethanol production by Kluyveromyces marxianus IMB3 during growth on straw-supplemented whiskey distillery spent wash at 45 °C

The thermotolerant, ethanol-producing yeast strain, Kluyveromyces marxianus IMB3 was grown on media consisting of straw-supplemented distillery spent wash from The Old Bushmill's Distillery Co. Ltd., Bushmills, Co Antrim, Northern Ireland. Media were supplemented with cellulase activity and fermentations were carried out at 45?°C. When pulverized straw was used as substrate in this system at concentrations of 2, 4 and 6% (w/v), ethanol concentrations increased to maxima of 1.45, 2.2 and 3?g/l, respectively. Based on straw containing a maximum of 40% cellulose, these ethanol concentrations accounted for 36, 27 and 24% of the maximum theoretical yield, respectively. When the straw was pre-treated with NaOH and used in the spent wash containing system at concentrations of 2, 4 and 6% (w/v) ethanol, concentrations increased to maxima of 3, 6.2 and 10.5?g/l, respectively and these accounted for 75, 76 and 86% of the maximum theoretical yield. When these results are compared with previously published data relating to the use of straw in laboratory-based media, they suggest that whiskey distillery spent wash may provide an adequate medium for supplementation with complex carbohydrate and subsequent ethanol production in simultaneous saccharification and fermentation processes.     

Response surface methodology for optimization of medium for decolorization of textile dye Direct Black 22 by a novel bacterial consortium

Decolorization and degradation of polyazo dye Direct Black 22 was carried out by distillery spent wash degrading mixed bacterial consortium, DMC. Response surface methodology (RSM) involving a central composite design (CCD) in four factors was successfully employed for the study and optimization of decolorization process. The hyper activities and interactions between glucose concentration, yeast extract concentration, dye concentration and inoculum size on dye decolorization were investigated and modeled. Under optimized conditions the bacterial consortium was able to decolorize the dye almost completely (>91%) within 12h. Bacterial consortium was able to decolorize 10 different azo dyes. The optimum combination of the four variables predicted through RSM was confirmed through confirmatory experiments and hence this bacterial consortium holds potential for the treatment of industrial waste water. Dye degradation products obtained during the course of decolorization were analyzed by HPTLC.     

Biodegradation of organic compounds of molasses melanoidin (MM) from biomethanated distillery spent wash (BMDS) during the decolourisation by a potential bacterial consortium

Molasses melanoidin (MM) is a major pollutant in biomethanated distillery spent wash (BMDS) due to its recalcitrant properties. The 75% colour and 71% COD of MM (1,000?ppm) were reduced with developed bacterial consortium comprising Proteus mirabilis (IITRM5; FJ581028), Bacillus sp. (IITRM7; FJ581030), Raoultella planticola (IITRM15; GU329705) and Enterobacter sakazakii (IITRM16, FJ581031) in the ratio of 4:3:2:1 within 10?days at optimized nutrient. Bacterial consortium showed manganese peroxidase and laccase activity during MM decolourisation. The dominant growth of R. planticola and E. sakazakii was noted in consortium during MM decolourisation. The comparative GC-MS analysis of extracted compounds of control and degraded samples showed that most of the compounds present in control were completely utilized by bacterial consortium along with production of some metabolites. The developed bacterial consortium could be a tool for the decolourisation and degradation of melanoidin containing BMDS.     

Decolorization and biodegradation of triphenylmethane dye,brilliant green,by Aspergillus sp. isolated from Ladakh,India

Brilliant green, used extensively to color silk and wool in the commercial textile industry is a hazardous recalcitrant. Aspergillus sp. strain CB-TKL-1 isolated from a water sample from Tsumoriri Lake, Karzok, Ladakh, India, was found to completely decolorize this dye within 72 h when cultured under aerobic conditions at 25 °C. The extent of decolorization was monitored by the decrease in absorbance maxima of the dye by UV–visible spectroscopy. The decolorization was optimum at pH 5 and 35 °C when agitated at 200 rpm. Addition of glucose (2%) as a carbon source and sodium nitrate (0.2%) as a nitrogen source enhanced the decolorization ability of the culture. The culture exhibited maximum extent of decolorization of brilliant green with a C:N ratio of 2.5 after 72 h. Thirteen N-demethylated decolorized products of brilliant green were identified based on UV–visible spectroscopy, Fourier Transform Infrared (FT-IR) spectroscopy and liquid chromatography–electrospray ionization mass spectrometry (LC–ESI-MS) analysis at the end of 72 h before mineralization. The difference of the relative absorption peaks in the decolorized sample indicated a linear release of N-demethylated compounds, indicating a stepwise N-demethylation in the decolorization process.     

Characterization of technological features of dry yeast (strain I-7-43) preparation, product of electrofusion between Saccharomyces cerevisiae and Saccharomyces diastaticus, in industrial application

The aim of the study was to verify the technological usability and stability of biotechnological features of active dry distillery yeast preparation (strain I-7-43 with amylolytic abilities) applied to full-scale production of agricultural distillery. Various reduced doses of glucoamylase preparation (San-Extra L) were used for starch saccharification, from 90% to 70% in relation to the full standard dose of preparation. The dry distillery yeast I-7-43 were assessed positively in respect to fermentation activity and yield of ethanol production. Application of the dry yeast I-7-43 preparation in distillery practice lowers the costs of spirit production by saving the glucoamylase preparation (up to 30%) used in the process of mash saccharification. Concentrations of the volatile fermentation by-products in raw spirits obtained from fermentations with application of I-7-43 strain were on the levels guaranteeing good organoleptic properties of distillates.     

Isolation and growth of the phototrophic bacterium Rhodopseudomonas palustris strain B1 in sago-starch-processing wastewater

An indigenous strain of the purple non-sulphur phototrophic bacterium, Rhodopseudomonas palustris strain B1, was selected for the utilization and treatment of wastewater from a sago-starch-processing decanter. Growth of Strain B1 under anaerobic–light conditions in the carbohydrate-rich effluent was optimized by using 50% (v/v) effluent diluted in a basal minimal mineral medium with the addition to 0.1% (w/v) yeast extract. The optimum level of nitrogen source supplement, ammonium sulphate, was 1.0g/l. Highest cell mass concentration was achieved by using tungsten lamps as the light source with a light intensity of 4 klux. Under these optimal conditions, a maximum biomass of about 2.5g dry cell/l with a pigment content of about 1.1mg carotenoid/g dry weight cell was achieved after 96h of anaerobic cultivation. There was a 77% reduc n the chemical oxygen demand (COD) of the effluent. A cell yield of about 0.59g dry weight cell/g COD was obtained.     

Decolourisation of synthetic and spentwash melanoidins using the white-rot fungus Phanerochaete chrysosporium JAG-40

Phanerochaete chrysosporium JAG-40 was isolated from the soil samples saturated with spilled molasses collected from a sugar mill. This isolate decolourised synthetic and natural melanoidins present in spentwash in liquid fermentation; up to 80% in 6 days at 30 degrees C under aerobic conditions. A large inoculum size stimulated fungal biomass production, but this gave less decolourisation of pigment; 5% w/v (dry weight) mycelial suspension was found optimum for maximum decolourisation in melanoidin medium supplemented with glucose and peptone. Gel-filtration chromatography showed that larger molecular weight fractions of melanoidin were decolourised more rapidly than small molecular weight fractions.     

Isolation of fungi and optimization of process parameters for decolorization of distillery mill effluent

Five different fungi isolated from distillery mill site in which two isolates (DF3 and DF4) had higher capabilities to remove color were identified as Emericella nidulans var. lata and Neurospora intermedia, respectively. Optimization of process parameter for decolorization was initially performed to select growth factors which were further substantiated by Taguchi approach in which seven factors, %carbon, %nitrogen, duration, pH, temperature, stirring and inoculum size, at two levels applying L-8 orthogonal array were taken for both fungi. Maximum color was removed at pH 3, temperature 30°C, stirring 125 rpm, dextrose (0.05%) and sodium nitrate (0.025%) by both fungi. After optimization, there was two-fold increase in color removal from 38 to 62% (DF3) and 31 to 64% (DF4) indicating significance of Taguchi approach in decolorization of distillery mill effluent. The mechanism for decolorization was determined by enzyme analysis, laccase and glucose oxidase, and indicated significant activity in DF3 as compared to DF4.     

Decolorization of an industrial effluent by free and immobilized cells of <Emphasis Type="Italic">Stenotrophomonas maltophilia</Emphasis> AAP56<Emphasis Type="Italic">.</Emphasis> Implementation of efficient down flow column reactor

A bacterial strain AAP56, isolated from a polluted soil (from Kelibia city) and identified as Stentrophomonas maltophilia, was particularly interesting for its ability to decolorize recalcitrant dyes of an industrial effluent: SITEX Black. The final percentage decolorization 60% was shown by bacterial culture after incubation in LB medium at 30°C under shaking conditions. The decolorization was closely correlated with the metabolic bacterial growth. The replacement of yeast extract in LB medium composition by soya flour was clearly efficient to enhance the percentage decolorization by 20% and also to reduce the growth medium cost 60-fold. The bacteria were able to reduce 23% from the initial COD and 28% from the initial BOD₅ of the effluent. The immobilization of bacterial cells in calcium alginate beads improved by 25% the effectiveness of the biotransformation within 24 h in batch conditions. The potential of a downflow fixed column reactor (DFCR) to decolorize SITEX Black was evaluated under dilution rate. The best decolorization percentage (82%) was recorded at 0.3 h⁻¹. This bioprocess seems to be a potentially useful method to remediate the colored textile wastewater.