Docosahexaenoic acid (DHA, C22:6n-3) plays a vital role in the enhancement of human health, particularly for cognitive, neurological, and visual functions. Marine microalgae, such as members of the genus Aurantiochytrium, are rich in DHA and represent a promising source of omega-3 fatty acids. In this study, levels of glucose, yeast extract, sodium glutamate and sea salt were optimized for enhanced lipid and DHA production by a Malaysian isolate of thraustochytrid, Aurantiochytrium sp. SW1, using response surface methodology (RSM). The optimized medium contained 60 g/L glucose, 2 g/L yeast extract, 24 g/L sodium glutamate and 6 g/L sea salt. This combination produced 17.8 g/L biomass containing 53.9% lipid (9.6 g/L) which contained 44.07% DHA (4.23 g/L). The optimized medium was used in a scale-up run, where a 5 L bench-top bioreactor was employed to verify the applicability of the medium at larger scale. This produced 24.46 g/L biomass containing 38.43% lipid (9.4 g/L), of which 47.87% was DHA (4.5 g/L). The total amount of DHA produced was 25% higher than that produced in the original medium prior to optimization. This result suggests that Aurantiochytrium sp. SW1 could be developed for industrial application as a commercial DHA-producing microorganism.
Composting of Palm Oil Mill Sludge (POMS) with sawdust was conducted in natural aerated reactor. Composting using natural aerated reactor is cheap and simple. The goal of this study is to observe the potential of composting process and utilizing compost as media for growing Cymbopogun citratus, one of Malaysia herbal plant. The highest maximum temperature achieved is about 40 degrees C and to increase temperature bed, more biodegradable substrate needs to be added. The pH value decrease along the process with final pH compost is acidic (pH 5.7). The highest maximum organic losses are about 50% with final C/N ratio of the compost is about 19. Final compost also showed some fertilizing value but need to be adjusted to obtain an ideal substrate. Addition of about 70% sandy soil causes highest yield and excellent root development for C. citratus in potted media. Beside that, compost from POMS-sawdust also found to have fertilizer value and easy to handle. Composting of POMS with sawdust shows potential as an alternative treatment to dispose and recycle waste components.
Photo fermentation is a biological process that can be applied for hydrogen production. The process is environmental friendly which is operated under mild conditions using renewable resources. In order to increase yield of H2 produced by Rhodobacter sphaeroides, some experimental factors that may enhance H2 production were studied. The effect of operating parameters including agitation, aeration and light on hydrogen production using R. sphaeroides NCIMB 8253 was investigated. Rhodobacter sphaeroides NCIMB 8253 was grown in 100 mL serum bottle containing growth medium with maliec acid as the sole organic carbon source. The cultures were incubated anaerobically at 30 degrees C with tungsten lamp (100 W) as the light source (3.8 klux) and argon gas was purged for maintaining anaerobic condition. The results show that maximum hydrogen produced was higher (54.37 mL) in static culture with 69.98% of H2 in the total gas compared with shake culture (11.57 mL) with 57.86% of H2. By using static culture, H2 produced was five times higher compared with non-static in both aerobic and anaerobic condition. It was found that growth and H2 production with fluorescent lamp showed better results than growth and H2 production with tungsten light.
Rice bran (RB) and de-oiled rice bran (DRB) have been treated and used as the carbon source in acetone-butanol-ethanol (ABE) production using Clostridium saccharoperbutylacetonicum N1-4. The results showed that pretreated DRB produced more ABE than pretreated RB. Dilute sulfuric acid was the most suitable treatment method among the various pretreatment methods that were applied. The highest ABE obtained was 12.13 g/L, including 7.72 g/L of biobutanol, from sulfuric acid. The enzymatic hydrolysate of DRB (ESADRB), when treated with XAD-4 resin, resulted in an ABE productivity and yield of 0.1 g/L h and 0.44 g/g, respectively. The results also showed that the choice of pretreatment method for RB and DRB is an important factor in butanol production.
The aim of this study was to investigate the influence of some environmental factors on bacterial metabolism. Fermentative hydrogen production by C. acetobutylicum, using glucose as the substrate. The effect of initial pH (4-8), inoculum size (1-20% (v/v)) and glucose concentration (1-30 g L(-1)) on hydrogen production were studied. The optimum cultivation temperature for hydrogen production was at 30 degrees C. The results show that substrate concentration and inoculum size resulted in hydrogen yield (Y(P/S)) of 391 mL g(-1) glucose utilized with maximum hydrogen productivity of 77.5 mL/L/h. Higher substrate concentration or inoculum size adversely affects hydrogen production, which decreases hydrogen yield by 15% to 334 mL g(-1) glucose utilized when 30% (v/v) inoculum size was used. The use of 30 g L(-1) substrate concentration resulted in a 75% decrease to 97 mL g(-1) glucose supplied. Concluded that proper Xo/So enhanced the hydrogen production.
The effect of removal of resultant gas resulted in enhancement of the H2 yield. The technique of CO2 scavenging resulted in H2 yield being improved from 408 mL g(-1) to reach the maximum of 422 mL g'. The highest hydrogen productivity of 87.9 ml L(-1) h(-1) was obtained by CO2 scavenging. Biomass concentration was enhanced to 1.47 g L(-1), Y(P,X) of 287 ml g(-1) L(-1), Y(X/S) of 0.294 and Y(H2/s) of 0.0377 by the use of CO2 scavenging. The results suggested that the presence of the gaseous products in fermentation medium and headspace adversely effect biomass growth and hydrogen production.
The objective of this study is to investigate the effect of salts addition to fermentation medium on hydrogen production, under anaerobic batch culture system. In this study, batch experiments were conducted to investigate the inhibitory effect of both NaCl and sodium acetate on hydrogen production. The optimum pH and temperature for hydrogen production were at initial pH of 7.0 and 30 degrees C. Enhanced production of hydrogen, using glucose as substrate was achieved. In the absence of Sodium Chloride and Sodium Acetate enhanced hydrogen yield (Y(P/S)) from 350 mL g(-1) glucose utilized to 391 mL g(-1) glucose utilized with maximum hydrogen productivity of 77.5 ml/L/h. Results also show that sodium chloride and sodium acetate in the medium adversely affect growth. Hydrogen yield per biomass (Y(P/X)) of 254 ml/L/g, biomass per substrate utilized (Y(X/S)) of 0.268 and (Y(H2/S) of 0.0349. The results suggested that Sodium at any concentration resulted to inhibit the bacterial productivity of hydrogen.
Date fruit juice contains high concentration of simple sugars ranging from 65 to 75% (w/w) in dry form. In this study, the potential of date fruit juice as biobutanol fermentation medium by C. acetobutylicum was investigated. The fermentation process was carried out at initial pH of 5, 6 and 7, incubation temperature of 30, 35 and 40 degrees C for 72 hours. The date fruit concentrations tested were 10, 20, 30 and 40 g L(-1). Medium containing 30 g L(-1) of date fruit at 35 degrees C incubation temperature with initial medium pH 7.0 gave the highest concentration of solvents of 3.1, 0.1 and 1.1 g L(-1) butanol, ethanol and acetone respectively. The yield and productivity of biobutanol were 0.32 g g(-1) and 0.044 g L(-1)/h respectively, while for total ABE were 0.45 g g(-1) and 0.06 g L(-1) h, respectively.
In this study, optimization of growth, lipid and DHA production of Aurantiochytrium SW1 was carried out using response surface methodology (RSM) in optimizing initial fructose concentration, agitation speed and monosodium glutamate (MSG) concentration. Central composite design was applied as the experimental design and analysis of variance (ANOVA) was used to analyze the data. ANOVA analysis revealed that the process which adequately represented by quadratic model was significant (p
In this work, hydrolysis of cellulose and hemicellulose content of palm kernel cake (PKC) by different types of hydrolytic enzymes was studied to evaluate monomeric sugars released for production of biobutanol by Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564) in acetone-butanol-ethanol (ABE) fermentation. Experimental results revealed that when PKC was hydrolyzed by mixed β-glucosidase, cellulase and mannanase, a total simple sugars of 87.81±4.78 g/L were produced, which resulted in 3.75±0.18 g/L butanol and 6.44±0.43 g/L ABE at 168 h fermentation. In order to increase saccharolytic efficiency of enzymatic treatment, PKC was pretreated by liquid hot water before performing enzymatic hydrolysis. Test results showed that total reducing sugars were enhanced to 97.81±1.29 g/L with elevated production of butanol and ABE up to 4.15±1.18 and 7.12±2.06 g/L, respectively which represented an A:B:E ratio of 7:11:1.
The locally isolated filamentous fungus Cunninghamella bainieri 2A1 was cultivated in a 5 L bioreactor to produce lipid and gamma-linolenic acid (GLA). The optimization was carried out using response surface methodology based on a central composite design. A statistical model, second-order polynomial model, was adjusted to the experimental data to evaluate the effect of key operating variables, including aeration rate and agitation speed on lipid production. Process analysis showed that linear and quadratic effect of agitation intensity significantly influenced lipid production process (P < 0.01). The quadratic model also indicated that the interaction between aeration rate and agitation speed had a highly significant effect on lipid production (P < 0.01). Experimental results showed that a lipid content of 38.71% was produced in optimum conditions using an airflow rate and agitation speed of 0.32 vvm and 599 rpm, respectively. Similar results revealed that 0.058(g/g) gamma-linolenic acid was produced in optimum conditions where 1.0 vvm aeration rate and 441.45 rpm agitation rate were used. The regression model confirmed that aeration and agitation were of prime importance for optimum production of lipid in the bioreactor.
The biosynthesis of biomedical products including lipid and gamma-linolenic acid (GLA) by Cunninghamella bainieri 2A1 was studied in repeated batch fermentation. Three key process variables, namely, glucose concentration, ammonium tartrate concentration, and harvesting time, were optimized using response surface methodology. Repeated batch fermentation was carried out by the cultivation of Cunninghamella bainieri 2A1 in nitrogen-limited medium with various nitrogen concentration (1-4 g/L) and glucose concentration (20-40 g/L) at three time intervals (12 h, 24 h, and 48 h). Experimental results showed that the highest lipid concentration of 6.2 g/L and the highest GLA concentration of 0.4 g/L were obtained in optimum conditions, where 20.2 g/L glucose, 2.12 g/L ammonium tartrate, and 48 h harvesting time were utilized. Statistical results showed that the interaction between glucose and ammonium tartrate concentration had highly significant effects on lipid and GLA biosynthesis (P < 0.01). Moreover, harvesting time had a significant interaction effect with glucose and ammonium tartrate concentration on lipid production (P < 0.05).
Palm kernel cake (PKC) was used for biobutanol production by Clostridium saccharoperbutylacetonicum N1-4 in acetone-butanol-ethanol (ABE) fermentation. PKC was subjected to acid hydrolysis pretreatment and hydrolysates released were detoxified by XAD-4 resin. The effect of pH, temperature and inoculum size on butanol production was evaluated using an empirical model. Twenty ABE fermentations were run according to an experimental design. Experimental results revealed that XAD-4 resin removed 50% furfural and 77.42% hydroxymethyl furfural. The analysis of the empirical model showed that linear effect of inoculums size with quadratic effect of pH and inoculum size influenced butanol production at 99% probability level (P<0.01). The optimum conditions for butanol production were pH 6.28, temperature of 28°C and inoculum size of 15.9%. ABE fermentation was carried out under optimum conditions which 0.1g/L butanol was obtained. Butanol production was enhanced by diluting PKC hydrolysate up to 70% in which 3.59g/L butanol was produced.
The production of biobutanol was studied by the cultivation of Clostridium acetobutylicum NCIMB 13557 in P2 medium including date fruit as the sole substrate. The effect of P2 medium and the effect of different concentrations of date fruit ranging from 10 to 100 g/L on biobutanol production were investigated. Anaerobic batch culture was carried out at 35 °C incubation temperature and pH 7.0 ± 0.2 for 72 h. Experimental results showed that the lowest yield of biobutanol and acetone-butanol-ethanol (ABE) was 0.32 and 0.35 gram per gram of carbohydrate consumed (g/g), respectively, when an initial date fruit concentration of 10 g/L was utilized. At this fruit date concentration a biobutanol production value of 1.56 g/L was obtained. On the other hand, the maximum yield of biobutanol (0.48 g/g) and ABE (0.63 g/g) was produced at 50 g/L date fruit concentration with a biobutanol production value as high as 11 g/L. However, when a higher initial date fruit concentration was used, biobutanol and ABE production decreased to reach the yield of 0.22 g/g and 0.35 g/g, respectively, where 100 g/L date fruit was used. Similar results also revealed that 10.03 g/L biobutanol was produced using 100 g/L date fruit.
Improvement in the butanol production selectivity or enhanced butanol:acetone ratio (B:A) is desirable in acetone-butanol-ethanol (ABE) fermentation by Clostridium strains. In this study, artificial electron carriers were added to the fermentation medium of a new isolate of Clostridium acetobutylicum YM1 in order to improve the butanol yield and B:A ratio. The results revealed that medium supplementation with electron carriers changed the metabolism flux of electron and carbon in ABE fermentation by YM1. A decrease in acetone production, which subsequently improved the B:A ratio, was observed. Further improvement in the butanol production and B:A ratios were obtained when the fermentation medium was supplemented with butyric acid. The maximum butanol production (18.20 ± 1.38 g/L) was gained when a combination of methyl red and butyric acid was added. Although the addition of benzyl viologen (0.1 mM) and butyric acid resulted in high a B:A ratio of 16:1 (800% increment compared with the conventional 2:1 ratio), the addition of benzyl viologen to the culture after 4 h resulted in the production of 18.05 g/L butanol. Manipulating the metabolic flux to butanol through the addition of electron carriers could become an alternative strategy to achieve higher butanol productivity and improve the B:A ratio.
Catalytic depolymerization of mannan composition of palm kernel cake (PKC) by mannanase was optimized to enhance the release of mannan-derived monomeric sugars for further application in acetone-butanol-ethanol (ABE) fermentation. Efficiency of enzymatic hydrolysis of PKC was studied by evaluating effects of PKC concentration, mannanase loading, hydrolysis pH value, reaction temperature and hydrolysis time on production of fermentable sugars using one-way analysis of variance (ANOVA). The ANOVA results revealed that all factors studied had highly significant effects on total sugar liberated (P<0.01). The optimum conditions for PKC hydrolysis were 20% (w/v) PKC concentration, 5% (w/w) mannanase loading, hydrolysis pH 4.5, 45°C temperature and 72h hydrolysis time. Enzymatic experiments in optimum conditions revealed total fermentable sugars of 71.54±2.54g/L were produced including 67.47±2.51g/L mannose and 2.94±0.03g/L glucose. ABE fermentation of sugar hydrolysate by Clostridium saccharoperbutylacetonicum N1-4 resulted in 3.27±1.003g/L biobutanol.
Continuous fermentation of dilute acid-pretreated de-oiled rice bran (DRB) to butanol by the Clostridium acetobutylicum YM1 strain was investigated. Pretreatment of DRB with dilute sulfuric acid (1%) resulted in the production of 42.12 g/L total sugars, including 25.57 g/L glucose, 15.1 g/L xylose and 1.46 g/L cellobiose. Pretreated-DRB (SADRB) was used as a fermentation medium at various dilution rates, and a dilution rate of 0.02 h-1 was optimal for solvent production, in which 11.18 g/L of total solvent was produced (acetone 4.37 g/L, butanol 5.89 g/L and ethanol 0.92 g/L). Detoxification of SADRB with activated charcoal resulted in the high removal of fermentation inhibitory compounds. Fermentation of detoxified-SADRB in continuous fermentation with a dilution rate of 0.02 h-1 achieved higher concentrations of solvent (12.42 g/L) and butanol (6.87 g/L), respectively, with a solvent productivity of 0.248 g/L.h. This study showed that the solvent concentration and productivity in continuous fermentation from SADRB was higher than that obtained from batch culture fermentation. This study also provides an economic assessment for butanol production in continuous fermentation process from DRB to validate the commercial viability of this process.
The effect of pH and butyric acid supplementation on the production of butanol by a new local isolate of Clostridium acetobutylicum YM1 during batch culture fermentation was investigated. The results showed that pH had a significant effect on bacterial growth and butanol yield and productivity. The optimal initial pH that maximized butanol production was pH 6.0 ± 0.2. Controlled pH was found to be unsuitable for butanol production in strain YM1, while the uncontrolled pH condition with an initial pH of 6.0 ± 0.2 was suitable for bacterial growth, butanol yield and productivity. The maximum butanol concentration of 13.5 ± 1.42 g/L was obtained from cultures grown under the uncontrolled pH condition, resulting in a butanol yield (YP/ S ) and productivity of 0.27 g/g and 0.188 g/L h, respectively. Supplementation of the pH-controlled cultures with 4.0 g/L butyric acid did not improve butanol production; however, supplementation of the uncontrolled pH cultures resulted in high butanol concentrations, yield and productivity (16.50 ± 0.8 g/L, 0.345 g/g and 0.163 g/L h, respectively). pH influenced the activity of NADH-dependent butanol dehydrogenase, with the highest activity obtained under the uncontrolled pH condition. This study revealed that pH is a very important factor in butanol fermentation by C. acetobutylicum YM1.
A new isolate of the solvent-producing Clostridium acetobutylicum YM1 was used to produce butanol in batch culture fermentation. The effects of glucose concentration, butyric acid addition and C/N ratio were studied conventionally (one-factor-at-a-time). Moreover, the interactions between glucose concentration, butyric acid addition and C/N ratio were further investigated to optimize butanol production using response surface methodology (RSM). A central composite design was applied, and a polynomial regression model with a quadratic term was used to analyze the experimental data using analysis of variance (ANOVA). ANOVA revealed that the model was highly significant (p
A two-stage fermentation process consisting of dark and photo-fermentation periods was carried out in a batch reactor. In the first stage, glucose was fermented in the dark stage using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564; CSN1-4) to produce acetate, CO2 and H2. The acetate produced in the first stage is fermented to H2 and CO2 by Rhodobacter sphaeroides NCIMB 8253 for further hydrogen production in the second, illuminated stage. The yield of hydrogen in the first stage was about 3.10 mol H2 (mol glucose)(-1) at a glucose concentration of 10 g L(-1), pH 6 +/- 0.2 and 37 degrees C and the second stage yield was about 1.10-1.25 mol H2 (mol acetic acid)(-1) at pH 6.8 +/- 0.2 and 32 degrees C, without removal of the Clostridium CSN1-4. The overall yield of hydrogen in the two-stage process, with glucose as the main substrate was higher than single-stage fermentation.