Enhanced Butanol Production Using Non-ionic Surfactant-Based Extractive Fermentation: Effect of Substrates and Immobilization of Cell.

The foremost aim of the present study was to enhance butanol production in an extractive fermentation study in presence of non-ionic surfactant using immobilized cells. Earlier studies had shown improved butanol production with non-ionic surfactant and immobilized cells independently. Therefore, in the present work, the combined effect of extractive fermentation and immobilized cells on butanol production was studied. Different matrices (brick, bamboo, cotton fiber, flannel cloth, and polyurethane foam) were tested for immobilization of Clostridium sporogenes. Immobilized biomass thus obtained was used in an extractive fermentation study with non-ionic surfactant L62. Biomass immobilized on polyurethane foam (PF) doubled the butanol production in presence of 6% (v/v) L62 with respect to control (free cells without surfactant). Further, the effect of different carbon sources was also studied to check the suitability of different industrial wastes containing different carbon sources. Glucose was found to be the best carbon source.

Effect of Operating Conditions and Immobilization on Butanol Enhancement in an Extractive Fermentation Using Non-ionic Surfactant.

The present study was undertaken in order to investigate effect of diverse parameters such as fermentation media, pH, initial concentration of biomass, different surfactant concentrations, and immobilization on increasing butanol and total solvent production. Cheng's fermentation media was successfully tested and perceived to increase final solvents concentration. Controlled pH at 12th and 24th hours had negative effect on butanol enhancement; however, it resulted in more butyric acid production which remained accumulated. Ten percent (v/v) biomass was evaluated to increase final solvents concentration and hence butanol yield compared to 20% and 30% (v/v) of initial biomass concentrations. Effect of surfactant concentration (3-20%) was studied on butanol production. Six percent (v/v) L62 resulted in 49% higher final butanol concentration compared to control. Simultaneous immobilization and fermentation showed higher butanol production (16.8 g/L with 6%) which was attributed to partial immobilization of biomass.

Enhanced n-butanol production by Clostridium beijerinckii MCMB 581 in presence of selected surfactant.

Extractive butanol fermentation with non-ionic surfactant, a recently explored area, has shown promising results with several advantages but is relatively less investigated. This work reports the extractive fermentation with selected non-ionic surfactants (L62 and L62D) to enhance butanol production using a high-butanol producing strain (Clostridium beijerinckii MCMB 581). Biocompatibility studies with both the surfactants showed growth. Higher concentrations of surfactant (>5%) affected the cell count. 15.3 g L-1 of butanol and 21 g L-1 of total solvents were obtained with 3% (v/v) L62 which was respectively, 43% (w/w) and 55% (w/w), higher than control. It was found that surfactant addition at 9th h doubled the productivity (from 0.13 to 0.31 g L-1 h-1 and 0.17 to 0.39 g L-1 h-1, respectively for butanol and total solvent). Butanol productivity obtained was 2-3 times higher than similar studies on extractive fermentation with non-ionic surfactants. Interestingly, mixing did not improve butanol production.

Xylitol production from non-detoxified and non-sterile lignocellulosic hydrolysate using low-cost industrial media components.

Immobilized Candida tropicalis cells in freeze dried calcium alginate beads were used for production of xylitol from lignocellulosic waste like corn cob hydrolysate without any detoxification and sterilization of media. Media components for xylitol fermentation were screened by statistical methods. Urea, KH2PO4 and initial pH were identified as significant variables by Plackett-Burman (PB) design. Significant medium components were optimized by response surface methodology (RSM). Predicted xylitol yield by RSM model and experimental yield was 0.87 and 0.79 g/g, respectively. Optimized conditions (urea 1.5 g/L, KH2PO4 1.9 g/L, xylose 55 g/L, pH 6.7) enhanced xylitol yield by 32% and xylose consumption by twofold over those of basal media. In addition, the immobilized cells were reused five times at shake flask level with optimized medium without affecting the xylitol productivity and yield. Xylitol production was successfully scaled up to 7.5 L stirred tank reactor using optimized media. Thus, the optimized condition with non-detoxified pentose hydrolysate from corn cob lignocellulosic waste with minimal nutrients without any sterilization opens up the scope for commercialization of the process.

Enhanced xylitol production using immobilized Candida tropicalis with non-detoxified corn cob hemicellulosic hydrolysate.

This study reports an industrially applicable non-sterile xylitol fermentation process to produce xylitol from a low-cost feedstock like corn cob hydrolysate as pentose source without any detoxification. Different immobilization matrices/mediums (alginate, polyvinyl alcohol, agarose gel, polyacrylamide, gelatin, and κ-carrageenan) were studied to immobilize Candida tropicalis NCIM 3123 cells for xylitol production. Amongst this calcium alginate, immobilized cells produced maximum amount of xylitol with titer of 11.1 g/L and yield of 0.34 g/g. Hence, the process for immobilization using calcium alginate beads was optimized using a statistical method with sodium alginate (20, 30 and 40 g/L), calcium chloride (10, 20 and 30 g/L) and number of freezing-thawing cycles (2, 3 and 4) as the parameters. Using optimized conditions (calcium chloride 10 g/L, sodium alginate 20 g/L and 4 number of freezing-thawing cycles) for immobilization, xylitol production increased significantly to 41.0 g/L (4 times the initial production) with corn cob hydrolysate as sole carbon source and urea as minimal nutrient source. Reuse of immobilized biomass showed sustained xylitol production even after five cycles.

Screening of non-Ionic Surfactant for Enhancing Biobutanol Production.

This work deals with finding a suitable non-ionic surfactant which has high butanol capturing capacity and can be separated at a temperature close to room temperature and does not extract any intermediates or substrate (i.e., glucose). Importantly, it should be biocompatible, and its separation from the aqueous phase is not affected by other fermentation products. Hence, a pool of non-ionic Pluronic surfactants (L31, L61, L62D, L62LF, L62, L81, L92, L101, L121, L64, P65, P84, P104, P105) were selected for the study. Screening of the surfactant was done based on its hydrophile-lipophile balance (HLB) value, butanol capturing capacity (BCC), and cloud point temperature. Among the various surfactant investigated, L62D captured maximum amount of butanol (0.68 g/g of surfactant). Also, the cloud point temperature of L62D is close to room temperature (28.7 °C). Biocompatibility studies were carried out by conducting fermentation in presence of 3% L62D which resulted in 148% increase in butanol production as compared to control (without surfactant). Further, the fermentation products did not have strong influence on phase separation.

Denitrification of high strength nitrate waste from a nuclear industry using acclimatized biomass in a pilot scale reactor.

This work investigates the performance of acclimatized biomass for denitrification of high strength nitrate waste (10,000 mg/L NO3) from a nuclear industry in a continuous laboratory scale (32 L) and pilot scale reactor (330 L) operated over a period of 4 and 5 months, respectively. Effect of substrate fluctuations (mainly C/NO3-N) on denitrification was studied in a laboratory scale reactor. Incomplete denitrification (95-96 %) was observed at low C/NO3-N (≤2), whereas at high C/NO3-N (≥2.25) led to ammonia formation. Ammonia production increased from 1 to 9 % with an increase in C/NO3-N from 2.25 to 6. Complete denitrification and no ammonia formation were observed at an optimum C/NO3-N of 2.0. Microbiological studies showed decrease in denitrifiers and increase in nitrite-oxidizing bacteria and ammonia-oxidizing bacteria at high C/NO3-N (≥2.25). Pilot scale studies were carried out with optimum C/NO3-N, and sustainability of the process was checked on the pilot scale for 5 months.

Extraction of p-coumaric acid and ferulic acid using surfactant-based aqueous two-phase system.

Ferulic acid (FA) and p-coumaric acid (pCA) are high-value products that can be obtained by alkaline hydrolysis of lignocellulose. Present work explores the potential of surfactant-based cloud-point extraction (CPE) for FA and pCA extraction from corn cob hydrolysate. More than 90 % (w/w) extraction of both FA and pCA was achieved from model system with L92. The partition coefficient of FA and pCA in L92 aqueous phase system was 35 and 55, respectively. A significant enrichment (8-10-fold) of both FA and pCA was achieved in surfactant-rich phase. Furthermore, the downstream process volume was reduced by 10 to 13 times. Optimized conditions (5 % v/v L92 and pH 3.0) resulted into 85 and 89 % extraction of FA and p-CA, respectively, from alkaline corn cob hydrolysate. Biocompatibility tests were carried out for L92 for ethanol fermentation and found to be biocompatible. Thus, the new surfactant-based CPE system not only concentrated FA and pCA but also reduced the process volume significantly. Further, aqueous phase containing sugars can be used for ethanol fermentation.

Simultaneous removal of carbon and nitrate in an airlift bioreactor.

This paper presents the integrated removal of carbon (measured as chemical oxygen demand i.e. COD) and NO(x)-N by sequentially adapted sludge, studied in an airlift reactor (ALR). Simultaneous removal of COD and nitrate occurs by denitrification (anoxic) and oxidation (aerobic). Aerobic (riser) and anoxic (remaining part) conditions prevail in different parts of the reactor. Studies were carried out in a 42 L ALR operated at low aeration rate to maintain anoxic and aerobic conditions as required for denitrification and COD removal, respectively. The sludge was adapted sequentially to increasing levels of NO(x)-N and COD over a period of 45 days. Nitrate removal efficiency of the sludge increased due to adaptation and degraded 900 ppm NO(3)-N completely in 2h (initially the sludge could not degrade 100 ppm NO(3)-N). The performance of the adapted sludge was tested for the degradation of synthetic waste with COD/N loadings in the range of 4-10. The reduction of COD was significantly faster in the presence of NO(x)-N and was attributed to the availability of oxygen from NO(x)-N and distinct conditions in the reactor. This hypothesis was justified by the material balance of COD.

Denitrification of highly alkaline nitrate waste using adapted sludge.

Uranium extraction and regeneration of ion exchange resin generates concentrated nitrate effluents (typically 500 to 10,000 ppm NO(3)-N) that are highly alkaline in nature (pH 9.0 to 11.0). It is difficult to remove nitrate from such solutions using standard physiochemical and biological methods. This paper reports denitrification of such wastes using preadapted sludge (biomass), which was acclimatized to different influent pH (7.5 to 11.5) in a sequencing batch reactor (4 l) for 2 months. Performance of the developed consortia was studied under different pH (7.5 to 12). Biomass denitrified the synthetic wastewater containing 1,694 ppm NO(3)-N at a pH of 10.5. Decrease in nitrite build up was observed at higher pH, which differs from the reported results. Kinetic analysis of the data showed that specific rate of nitrate reduction was highest (78 mg NO(3)-N/g MLSS/h) at higher pH (10.5). This was attributed to the acclimatization process. Thus, high-strength nitrate wastewater, which was highly alkaline, was successfully treated using preadapted sludge.

Biotreatment of high strength nitrate waste using immobilized preadapted sludge.

One of the major wastes generated by fertilizer, explosive, and nuclear industries are nitrate (as high as 1,000 ppm NO(3)N) whose removal before disposal has become a growing concern. In this study, an active denitrifying sludge was immobilized onto support materials like cloth and polyurethane foam and their denitrification efficiency on high nitrate wastes [1,000 ppm NO(3) (225 ppm NO(3)N), 5,000 ppm NO(3) (1,129 ppm NO(3)N), 7,500 ppm NO(3) (1,693 ppm NO(3) N)] was studied. Results showed complete degradation of the nitrate wastes (225 ppm NO(3)N, 1,129 ppm NO(3)N, and 1,693 ppm NO(3)N) without any accumulation of nitrite in a period of only 1, 4, and 10 h, respectively. Based on adhering and entrapment principle, an immobilization unit was developed using a combination of cloth and foam as well as both individually. This system used for treating such high nitrate wastes was found to be quite effective in waste water treatment, particularly in problems associated with solid-liquid separation. The batch column reactor was run in about 45 batches without any loss in activity or reactor stability.

Biological denitrification of high strength nitrate waste using preadapted denitrifying sludge.

Denitrification of synthetic high nitrate waste containing 9032 ppm NO(3)-N (40,000 ppm NO(3)) in a time period of only 6h has been achieved in our previous study using activated sludge. The activated sludge culture was acclimatized by a stepwise increase in the nitrate concentration of synthetic waste. In the present work, studies were carried out on the changing microbial population of the sludge and the physiology of nitrate metabolism during the various stages of adaptation process to high strength synthetic nitrate waste. During the course of adaptation, with an increase in the nitrate concentration, a sharp increase in the number of denitrifiers was found with an equally rapid decrease in the nitrifying community. Two key enzymes involved in the first two steps of the denitrification process were also studied during this period. The results of the study suggest that specific enzyme levels increase as the activated sludge adapts itself to higher nitrate concentrations. Biological denitrification of high nitrate waste is a slow process and to increase the rate of denitrification, parameters such as pH, temperature, C:N and biomass concentration of the process were optimized using orthogonal array method. Optimized conditions increased the specific nitrate reduction rate by 54% and specific nitrite reduction rate by 45%.

Denitrification of high strength nitrate waste.

The aim of the present work was to study the treatment of high strength nitrate waste (40000 ppm NO(3) i.e., 9032 ppm NO(3)-N) by acclimatizing sludge initially capable of degrading dilute streams (100-200 ppm NO(3)-N). Sludge from an effluent treatment plant of a fertilizer industry was acclimatized for 15 d each at 1694, 3388, 6774 and 9032 ppm NO(3)-N in a 4 L sequencing batch reactor. Complete denitrification of extremely concentrated nitrate waste (9032 ppm NO(3)-N) using acclimatized sludge was achieved in just 6 h. During the acclimatization period, increase in nitrite peak value from zero to 5907 ppm NO(2)-N was observed, as the concentration was increased from 1694 to 9032 ppm NO(3)-N. Kinetic analysis of the nitrate and nitrite profile could reasonably support microbiological explanations for nitrite build up and changes in sludge composition.