Enzymatical hydrolysis of food waste and ethanol production from the hydrolysate

The aim of present paper was to investigate the prospect for the use of food waste, an important municipal waste, as a potential substrate to generate hydrolysates for fuel ethanol production. The critical variables that affected reducing sugar production from food waste were identified by Plackett–Burman design (glucoamylase loud, time, temperature and pH) and further optimized by using a four factor central composite design of response surface methodology. According to the results of response surface analysis, the optimum conditions for reducing sugar production were determined to be glucoamylase loud of 142.2 u/g, saccharification pH of 4.82, enzyme reaction temperature of 55 °C, enzyme reaction time of 2.48 h. Reducing sugar production (164.8 g/L) in the optimized condition was in good agreement with the value predicted by the quadratic model (164.3 g/L), thereby confirming its validity. Furthermore, the obtained liquid phase of food waste hydrolysate was utilized for production of ethanol by using Saccharomyces cerevisiae H058 fermentation. In order to develop an economical process for transforming food waste hydrolysates to ethanol, non-sterilized and sterilized processes were compared in the experiments. The result shows non-sterilized fermentation without undergoing heat treatment was better due to the unspoiled nutrients inside. These results helped to find the effective strategies to utilize food waste for ethanol production.     

Lactic acid recovery from fermentation broth of kitchen garbage by esterification and hydrolysis method

Kitchen garbage was utilized to produce lactic acid (LA) to reduce the corresponding cost. The whole process for pure LA production involved fermentation, esterification and hydrolysis. Kitchen garbage could produce 47.9 g L^?1 LA with pH adjusted with ammonia to 6–7. Then the fermentation broth was concentrated by water evaporation, the ammonium lactate inside was esterified with the butanol to produce butyl lactate. Proper catalyst was studied to improve esterification rate, a cation-exchange resin modified by FeCl₃ as a catalyst was proved to be effective. The esterification yield of ammonium lactate (NH₄LA) could reach 96%. Pure LA was hydrolyzed from the obtained butyl lactate in presence of a cation-exchange resin in the H⁺ form as a catalyst. The catalyst for hydrolysis could be regenerated and reused to save the cost. LA production from the kitchen garbage could not only save cost, but also solve the pollution problems of kitchen garbage.     

Optimization of ethanol production from hot-water extracts of sugar maple chips

Hot-water extracts from sugar maple chips prior to papermaking was employed in this study to produce ethanol by Pichia stipitis 58784. The effects of several factors, seed culture age, fermentation time, inoculum quantity, agitation rate, percent extract, concentration of inorganic nitrogen source (NH₄)₂SO₄ and pH value, on ethanol production were investigated by orthogonal experiments. Orthogonal analysis shows that the optimal fermentation was obtained in the condition of 48-h seed culture, 120-h fermentation, 16% inoculum, 180 rpm, containing 30% extracts, 8% ammonium sulphate supplement and pH 5. This optimal condition was verified at 800-mL level in a 1.3 L fermentor. The ethanol yield reached 82.27% of the theoretical (20.57 g/L) after 120 h.     

Biological hydrogen production from probiotic wastewater as substrate by selectively enriched anaerobic mixed microflora

Biohydrogen production from probiotic wastewater using mixed anaerobic consortia is reported in this paper. Batch tests are carried out in a 5.0 L batch reactor under constant mesophillic temperature (37 °C). The maximum hydrogen yield 1.8 mol-hydrogen/mol-carbohydrate is obtained at an optimum pH of 5.5 and substrate concentration 5 g/L. The maximum hydrogen production rate is 168 ml/h. The hydrogen content in the biogas is more than 65% and no significant methane is observed throughout the study. In addition to hydrogen, acetate, propionate, butyrate and ethanol are found to be the main by-products in the metabolism of hydrogen fermentation.     

Production of 16% ethanol from 35% sucrose

A strain of Saccharomyces cerevisiae, which showed marked fermentation activity, ethanol and temperature tolerance and good flocculation ability, was selected for ethanol production. A stuck fermentation occurred at sucrose concentration of 25%. Increasing the yeast inoculum volume from 3% to 6% showed positive effects on fermentation from 25% sucrose. The ratio of added nitrogen to sucrose, which gave the best results (for the selected yeast strain), was determined. It was concluded that this ratio (nitrogen as ammonium sulphate at a rate of 5 mg g^?1 of consumed sucrose) is constant at various sugar concentrations. Addition of nitrogen at this ratio produced 11.55% ethanol with complete consumption of 25% sucrose after 48 h of fermentation. However fermentation of 30% sucrose at the above optimum conditions was not complete. Addition of yeast extract at a level of 6 g l^?1 together with thiamine at a level of 0.2 g l^?1 led to complete utilization of 30% sucrose with resultant 14% ethanol production. However the selected yeast strain was not able to ferment 35% sucrose at the same optimum conditions. Addition of air at a rate of 150 dm³ min^?1 m³ of reactor volume during the first 12 h of fermentation led to complete consumption of 35% sucrose and 16% ethanol was produced. This was approximately the theoretical maximum for ethanol production.     

Cellulase production using biomass feed stock and its application in lignocellulose saccharification for bio-ethanol production

A major constraint in the enzymatic saccharification of biomass for ethanol production is the cost of cellulase enzymes. Production cost of cellulases may be brought down by multifaceted approaches which include the use of cheap lignocellulosic substrates for fermentation production of the enzyme, and the use of cost efficient fermentation strategies like solid state fermentation (SSF). In the present study, cellulolytic enzymes for biomass hydrolysis were produced using solid state fermentation on wheat bran as substrate. Crude cellulase and a relatively glucose tolerant BGL were produced using fungi Trichoderma reesei RUT C30 and Aspergillus niger MTCC 7956, respectively. Saccharification of three different feed stock, i.e. sugar cane bagasse, rice straw and water hyacinth biomass was studied using the enzymes. Saccharification was performed with 50 FPU of cellulase and 10 U of β-glucosidase per gram of pretreated biomass. Highest yield of reducing sugars (26.3 g/L) was obtained from rice straw followed by sugar cane bagasse (17.79 g/L). The enzymatic hydrolysate of rice straw was used as substrate for ethanol production by Saccharomyces cerevisiae. The yield of ethanol was 0.093 g per gram of pretreated rice straw.     

Optimization of media conditions for the production of ethanol from sweet sorghum juice by immobilized Saccharomyces cerevisiae

In order to obtain high ethanol yield and fermentation rate, response surface methodology (RSM) was employed to study the effect of culture medium on the ethanol productivity from stalk juice of sweet sorghum by immobilized yeast. A 2³ central composite design (CCD) was chosen to explain the combined effects of the medium constituents, viz. nitrogen (adjusted by adding (NH₄)₂SO₄), phosphorus (adjusted by adding KH₂PO₄), and pH. A mathematical correlation about the influence of the nitrogen, phosphorus, and pH on the ethanol productivity was established. It predicted that the maximum ethanol production rate (119.12 g/l h) was observed for a medium consisting of 0.77 g/l phosphorus, 2.15 g/l nitrogen, and pH = 6.39. Under this condition, the ethanol fermentation rate was 122.85 g/l h.     

Bioethanol production by a flocculent hybrid,CHFY0321 obtained by protoplast fusion between Saccharomyces cerevisiae and Saccharomyces bayanus

Fusion hybrid yeast, CHFY0321, was obtained by protoplast fusion between non-flocculent-high ethanol fermentative Saccharomyces cerevisiae CHY1011 and flocculent-low ethanol fermentative Saccharomyces bayanus KCCM12633. The hybrid yeast was used together with the parental strains to examine ethanol production in batch fermentation. Under the conditions tested, the fusion hybrid CHFY0321 flocculated to the highest degree and had the capacity to ferment well at pH 4.5 and 32 °C. Simultaneous saccharification and fermentation for ethanol production was carried out using a cassava (Manihot esculenta) powder hydrolysate medium containing 19.5% (w v^?1) total sugar in a 5 l lab scale jar fermenter at 32 °C for 65 h with an agitation speed of 2 Hz. Under these conditions, CHFY0321 showed the highest flocculating ability and the best fermentation efficiency for ethanol production compared with those of the wild-type parent strains. CHFY0321 gave a final ethanol concentration of 89.8 ± 0.13 g l^?1, a volumetric ethanol productivity of 1.38 ± 0.13 g l^?1 h^?1, and a theoretical yield of 94.2 ± 1.58%. These results suggest that CHFY0321 exhibited the fermentation characteristics of S. cerevisiae CHY1011 and the flocculent ability of S. bayanus KCCM12633. Therefore, the strong highly flocculent ethanol fermentative CHFY0321 has potential for improving biotechnological ethanol fermentation processes.     

Fuel ethanol production from steam-pretreated corn stover using SSF at higher dry matter content

Replacing fossil fuels by bio-fuels has many advantages, such as the reduction of CO₂-emission to the atmosphere, the possibility for non-oil-producing countries to be self-sufficient in fuel, and increased local job opportunities. Bio-ethanol is such a promising renewable fuel. However, today it is produced from sugar or starch—raw materials that are relatively expensive. To lower the production cost of bio-ethanol the cost of the raw material must be reduced and the production process made more efficient. The production of bio-ethanol from corn stover using simultaneous saccharification and fermentation (SSF) at high dry matter content addresses both issues. Corn stover is an agricultural by-product and thus has a low economic value. SSF at high dry matter content results in a high ethanol concentration in the fermented slurry, thereby decreasing the energy demand in the subsequent distillation step.In this study, SSF was performed on steam-pretreated corn stover at 5, 7.5 and 10% water-insoluble solids (WIS) with 2 g/L hexose-fermenting Saccharomyces cerevisiae (ordinary compressed baker's yeast). SSF at 10% WIS resulted in an ethanol yield of 74% based on the glucose content in the raw material and an ethanol concentration of 25 g/L. Neither higher yeast concentration (5 g/L) nor yeast cultivated on the liquid after the pretreatment resulted, under these conditions, in a higher overall ethanol yield.     

Carob pod as a feedstock for the production of bioethanol in Mediterranean areas

There is a growing interest worldwide to find out new and cheap carbohydrate sources for production of bioethanol. In this context, carob pod (Ceratonia siliqua) is proposed as an economical source for bioethanol production, especially, in arid regions. The carob tree is an evergreen shrub native to the Mediterranean region, cultivated for its edible seed pods and it is currently being reemphasised as an alternative in dryland areas, because no carbon-enriched lands are necessary. In this work, the global process of ethanol production from carob pod was studied. In a first stage, aqueous extraction of sugars from the pod was conducted, achieving very high yields (>99%) in a short period of time. The process was followed by acid or alkaline hydrolysis of washed pod at different operating conditions, the best results (R = 38.20%) being reached with sulphuric acid (2% v/v) at 90 °C, using a L/S (liquid/solid) ratio of 7.5 and shaking at 700 rpm for 420 min. After that, fermentation of hydrolysates were tested at 30 °C, 125 rpm, 200 g/L of sugars and 15 g/L of yeast with three different kinds of yeasts. In these conditions a maximum of 95 g/L of ethanol was obtained after 24 h. Finally, the distillation and dehydration of water–bioethanol mixtures was analyzed using the chemical process simulation software CHEMCAD with the aim of estimate the energy requirements of the process.     

Photocatalytic colour and COD removal in the distillery effluent by solar radiation

The photodegradation of distillery effluent has been studied for removal of colour and COD reduction in the presence of solar radiation. The influence of experimental parameters such as H₂O₂ concentration dosage, effluent COD concentration, TiO₂ catalyst and pH on colour and COD removal efficiency through solar photochemical process has been investigated. Maximum colour removal of the distillery effluent achieved was 79% at an H₂O₂ concentration of 0.3 M, pH 6, effluent COD concentration of 500 ppm and catalyst dosage of 0.1 g/L. The TiO₂/H₂O₂ system seems to be more efficient in comparison to the synergetic action that appears when using H₂O₂ and TiO₂. The photocatalytic degradation process using solar light as an irradiation source showed potential application for the colour removal of the distillery effluent treatment. Solar radiation can be an considered as an alternative, effective and economic energy carrier for the treatment of industrial effluent.     

Optimization of ethanol production from sweet sorghum (Sorghum bicolor) juice using response surface methodology

Sweet sorghum juice was fermented into ethanol using Saccharomyces cerevisiae (ATCC 24858). Factorial experimental design, regression analysis and response surface method were used to analyze the effects of the process parameters including juice solid concentration from 6.5 to 26% (by mass), yeast load from 0.5 g L⁻¹ to 2 g L⁻¹ and fermentation temperature from 30 °C to 40 °C on the ethanol yield, final ethanol concentration and fermentation kinetics. The fermentation temperature, which had no significant effect on the ethanol yield and final ethanol concentration, could be set at 35 °C to achieve the maximum fermentation rate. The yeast load, which had no significant effect on the final ethanol concentration and fermentation rate, could be set at 1 g L⁻¹ to achieve the maximum ethanol yield. The juice solid concentration had significant inverse effects on the ethanol yield and final ethanol concentration but a slight effect on the fermentation rate. The raw juice at a solid concentration of 13% (by mass) could be directly used during fermentation. At the fermentation temperature of 35 °C, yeast solid concentration of 1 g L⁻¹ and juice solid concentration of 13%, the predicted ethanol yield was 101.1% and the predicted final ethanol concentration was 49.48 g L⁻¹ after 72 h fermentation. Under this fermentation condition, the modified Gompertz's equation could be used to predict the fermentation kinetics. The predicted maximum ethanol generation rate was 2.37 g L⁻¹ h⁻¹.     

Simultaneous saccharification and fermentation (SSF) of very high gravity (VHG) potato mash for the production of ethanol

Simultaneous saccharification and fermentation (SSF) of very high gravity (VHG) potato mash, containing 304 g L^?1 of dissolved carbohydrates, was carried out for ethanol production. Potato tubers were ground into a mash, which was highly viscous. Mash viscosity was reduced by the pretreatment with mixed enzyme preparations of pectinase, cellulase and hemicellulase. The enzymatic pretreatment established the use of VHG mash with a suitable viscosity. Starch in the pretreated mash was liquefied to maltodextrins by the action of thermo-stable α-amylase at 85 °C. SSF of liquefied mash was performed at 30 °C with the simultaneous addition of glucoamylase, yeast (Saccharomyces cerevisiae) and ammonium sulfate as a nitrogen source for the yeast. The optimal glucoamylase loading, ammonium sulfate concentration and fermentation time were 1.65 AGU g^?1, 30.2 mM and 61.5 h, respectively, obtained using the response surface methodology (RSM). Ammonium sulfate supplementation was necessary to avoid stuck fermentation under VHG condition. Using the optimized condition, ethanol yield of 16.61% (v/v) was achieved, which was equivalent to 89.7% of the theoretical yield.     

Environmental aspects of ethanol derived from no-tilled corn grain: nonrenewable energy consumption and greenhouse gas emissions

Nonrenewable energy consumption and greenhouse gas (GHG) emissions associated with ethanol (a liquid fuel) derived from corn grain produced in selected counties in Illinois, Indiana, Iowa, Michigan, Minnesota, Ohio, and Wisconsin are presented. Corn is cultivated under no-tillage practice (without plowing). The system boundaries include corn production, ethanol production, and the end use of ethanol as a fuel in a midsize passenger car. The environmental burdens in multi-output biorefinery processes (e.g., corn dry milling and wet milling) are allocated to the ethanol product and its various coproducts by the system expansion allocation approach.The nonrenewable energy requirement for producing 1 kg of ethanol is approximately 13.4–21.5 MJ (based on lower heating value), depending on corn milling technologies employed. Thus, the net energy value of ethanol is positive; the energy consumed in ethanol production is less than the energy content of the ethanol (26.8 MJ kg⁻¹).In the GHG emissions analysis, nitrous oxide (N₂O) emissions from soil and soil organic carbon levels under corn cultivation in each county are estimated by the DAYCENT model. Carbon sequestration rates range from 377 to 681 kg C ha⁻¹ year⁻¹ and N₂O emissions from soil are 0.5–2.8 kg N ha⁻¹ year⁻¹ under no-till conditions. The GHG emissions assigned to 1 kg of ethanol are 260–922 g CO₂ eq. under no-tillage. Using ethanol (E85) fuel in a midsize passenger vehicle can reduce GHG emissions by 41–61% km⁻¹ driven, compared to gasoline-fueled vehicles. Using ethanol as a vehicle fuel, therefore, has the potential to reduce nonrenewable energy consumption and GHG emissions.     

Optimization and analysis of a bioethanol agro-industrial system from sweet sorghum

The use of non-food crops for bioethanol production represents an important trend for renewable energy in China. In this paper, a bioethanol agro-industrial system with distributed fermentation plants from sweet sorghum is presented. The system consists of the following processes: sweet sorghum cultivation, crude ethanol production, ethanol refining and by-product utilization. The plant capacities of crude ethanol and pure ethanol, in different fractions of useful land, are optimized. Assuming a minimum cost of investment, transport, operation and so on, the optimum capacity of the pure ethanol factory is 50,000 tonnes/year. Moreover, this bioethanol system, which requires ca. 13,300 ha (hectares) of non-cultivated land to supply the raw materials, can provide 26,000 jobs for rural workers. The income from the sale of the crops is approximately 71 million RMB Yuan and the ethanol production income is approximately 94 million RMB Yuan. The potential savings in CO₂ emissions are ca. 423,000 tonnes/year and clear economic, social and environmental benefits can be realized.     

Screening of microorganisms for xylitol production and fermentation behavior in high concentrations of xylose

Microorganisms with the ability to produce xylitol from high concentrations of xylose were screened from soils by enrichment culture using xylose as a sole carbon source. The selected strain was classified and determined as Candida sp. according to a taxonomic identification. In this strain, various conditions for xylitol production were investigated. Organic nutrients such as peptone and yeast extract were essential for xylitol production, and the optimal initial pH and k_La were between 4.0 and 6.0, and 5.2/h, respectively. Under the optimal condition, xylitol production from 200 g/l of xylose was 173 g/l after 5 days incubation, a 99.3% yield of the theoretical value. In order to produce a higher concentration of xylitol, a fed-batch culture of xylose was carried out by feeding 2.0 g of xylose per 10 ml of culture medium. The production from 4.0 g of xylose was 2.9 g of xylitol (concentration of 256 g/l) after 11.5 days which is a yield of 85.0%.     

Screening for sugar and ethanol processing characteristics from anatomical fractions of wheat stover

Due to concerns with stover collection systems, soil sustainability, and processing costs to produce ethanol, there are opportunities to investigate the optimal plant fractions to collect. Wheat stover fractions were separated by hand and analyzed for glucan, xylan, acid-soluble lignin, acid-insoluble lignin, and ash composition. Internodes had the highest glucan content (38.2% zero percent moisture basis) and the other fractions varied between 29.9% and 33.4%. The stover fractions were pretreated with either 0%, 0.4%, or 0.8% NaOH for 2 h at room temperature, washed, autoclaved, and saccharified. In addition, acid pretreated samples underwent simultaneous saccharification and fermentation (SSF) to ethanol. In general, the acid and alkaline pretreatments produced similar trends with leaves requiring very little pretreatment to achieve high conversion rates (greater than 80%). Chaff responded very well to pretreatment and high conversion efficiencies resulted when pretreated under alkaline or acidic conditions. Nodes and internodes were more recalcitrant than the other anatomical fractions. Pretreatment with 0.8% sulfuric acid (0.24 g sulfuric acid/g biomass) did not result in a significantly higher conversion of glucan to ethanol as the native material. Pretreatment with 0.8% NaOH (0.06 g NaOH/g biomass) at room temperature for 2 h resulted in high conversion efficiencies for all plant fractions, greater than 73% of the available glucan. These differences in pretreatment susceptibilities suggest that a biomass collection system that removes specific portions of wheat stover could result in significant differences in ethanol production costs.     

Effects of dark/light bacteria ratio on bio-hydrogen production by combined fed-batch fermentation of ground wheat starch

Bio-hydrogen production by combined dark and light fermentation of ground wheat starch was investigated using fed-batch operation. Serum bottles containing heat-treated anaerobic sludge and a mixture of Rhodobacter sp. was fed with a medium containing 20 g dm^?3 wheat powder (WP) at a constant flow rate. The system was operated at different initial dark/light biomass ratios (D/L). The optimum D/L ratio was 1/2 yielding the highest cumulative hydrogen (1548 cm³), yield (65.2 cm³ g^?1 starch), and specific hydrogen production rate (5.18 cm³ g^?1 h^?1). Light fermentation alone yielded higher hydrogen production than dark fermentation due to fermentation of volatile fatty acids (VFAs) to H₂ and CO₂. The lowest hydrogen formation was obtained with D/L ratio of 1/1 due to accumulation of VFAs in the medium.     

Alkalinity and high total solids affecting H2 production from organic solid waste by anaerobic consortia

The optimization of total solids in the feed (%TS) and alkalinity ratio (γ) for H₂ production from organic solid wastes under thermophilic regime was carried out using response surface methodology based on a central composite design. The total solids levels were 20.9, 23.0, 28.0, 33.0 and 35.1% whereas the levels of alkalinity ratio (defined as g phosphate alkalinity/g dry substrate) were 0.15, 0.20, 0.30, 0.41 and 0.45. High levels of TS and γ affected in a negative way the H₂ productivity and yield; both response variables significantly increased upon decreasing the TS content and alkalinity ratio. The highest H₂ productivity and yield were 463.7 N mL/kg-d and 54.8 N mL/g VS_rem, respectively, predicted at 20.9% TS and alkalinity ratio 0.25 (0.11 g CaCO₃/g dry substrate). The alkalinity requirements for hydrogenogenic processes were lower than those reported for methanogenic processes (0.11 vs. 0.30 g CaCO₃/g COD). Adequate alkalinity ratio was necessary to maintain optimal biological activity for hydrogen production; however, excessive alkalinity negatively affected process performance probably due to an increase of osmotic pressure. Interestingly, reactor pH depended only on the alkalinity ratio, thus the buffer capacity was able to maintain a constant pH independently of TS levels. At γ = 0.15–0.30 the pHs were in the range 5.56–5.95, which corresponded to the highest hydrogen productivities and yields. Finally, the highest metabolite accumulation corresponded with the highest removal efficiencies but not with high H₂ productivities and yields. Therefore, it seems that organic matter removal was channeled toward solvent generation instead of hydrogen production at high TS and γ levels. This is the first study that shows the requirements of alkalinity in solid substrate fermentation conditions for H₂ production processes and their interaction with the content of total solids in the feed.     

Ethanol production from hydrothermal pretreated corn stover with a loop reactor

Hydrothermal pretreatment on raw corn stover (RCS) with a loop reactor was investigated at 195 °C for different times varying between 10 min and 30 min. After pretreatment, the slurry was separated into water-insoluble solid (WIS) and liquid phase. Glucan and xylan were found in the both phases. The pretreatment condition showed a significant impact on xylan recovery. As the pretreatment time prolonged from 10 min to 30 min, the xylan recovery from liquid phase changed between 39.5% and 45.6% and the total xylan recoveries decreased from 84.7% to 61.6%. While the glucan recovery seemed not sensitive to the different pretreatment times. The glucan recovered from liquid was from 4.9% to 5.6% and the total glucan recoveries from all the pretreatments were higher than 98%. Besides HMF and furfural, acetic, lactic, formic and glycolic acids were also found in the liquid phase. All the concentrations of these potential inhibitors were lower enough not to affect the activity of the Saccharomyces cerevisiae (S. cerevisiae). Compared with the ethanol production of 32.4% from the RCS with S. cerevisiae, all the WISs gave higher ethanol productions ranging between 61.2% and 71.2%. When the xylan was taken into consideration, the best pretreatment condition would be 195 °C, 15 min and the estimated total ethanol production was 201 g kg^?1 RCS by assuming the fermentation of both C-6 and C-5 with the ethanol yield of 0.51 g g^?1 and 0.47 g g^?1, respectively.