Non-sterile simultaneous saccharification and fermentation of corn leaves and stalks to <Emphasis Type="SmallCaps">l</Emphasis>-lactic acid without external nutrient addition

Fermentation of lignocellulosic biomass requires auxiliary materials, including nutrients, to ensure the proliferation of microorganisms. Nutrients are usually inexpensive, but their contribution to the cost is considerable because of the very low prices of fermentation products, such as bio-ethanol. Using substances present in native lignocellulosic biomass as nutrients for fermentation was proposed and demonstrated. Leaves and stalks of corn plants were used as biomass, and nutrients were recovered as a nutrient solution by soaking them in water before alkaline peroxide pretreatment. Pretreated biomass and the nutrient solution derived from the same lot were used for non-sterile simultaneous enzymatic saccharification and thermophilic l-lactic acid fermentation (SSF). Using the nutrient solution in the saccharification step did not impact sugar recovery, and instead improved sugar yields because of the presence of eluted sugars in the solution. The l-lactic acid yield of 0.33 g g^?1 based on native biomass weight indicated that the nutrient solution functioned as a source of nutrients and sugars, especially as a source of essential phosphorus. Comparatively, autoclaved SSF yielded less or no l-lactic acid, indicating an apparent inhibitive effect derived from the nutrient solution on bacterial growth.     

Feasibility of producing ethanol from food waste

Food waste generated in Korea is rich in carbohydrate as high as 65% of total solids. Using the food waste, the feasibility of ethanol production was investigated in a lab-scale fermentor. Pretreatment with hydrolyzing enzymes including carbohydrase, glucoamylase, cellulase and protease were tested for hydrolysis of food waste. The carbohydrase was able to hydrolyze and produce glucose with a glucose yield of 0.63 g glucose/g total solid. Enzymatic hydrolysis and ethanol fermentation by using carbohydrase and Saccharomyces cerevisiae were conducted in the batch mode. For separated hydrolysis and fermentation (SHF), ethanol concentration reached at the level corresponding to an ethanol yield of 0.43 g ethanol/g total solids. For simultaneous saccharification and fermentation (SSF), the ethanol yield was 0.31 g ethanol/g total solids. During the continuous operation of SHF, the volumetric ethanol production rate was 1.18 g/lh with an ethanol yield of 0.3g ethanol/g total solids. For SSF process, the volumetric ethanol production rate was 0.8 g/lh with an ethanol yield of 0.2g ethanol/g total solids.     

Ethanol production from banana peels using statistically optimized simultaneous saccharification and fermentation process

Dried and ground banana peel biomass (BP) after hydrothermal sterilization pretreatment was used for ethanol production using simultaneous saccharification and fermentation (SSF). Central composite design (CCD) was used to optimize concentrations of cellulase and pectinase, temperature and time for ethanol production from BP using SSF. Analysis of variance showed a high coefficient of determination (R²) value of 0.92 for ethanol production. On the basis of model graphs and numerical optimization, the validation was done in a laboratory batch fermenter with cellulase, pectinase, temperature and time of nine cellulase filter paper unit/gram cellulose (FPU/g-cellulose), 72 international units/gram pectin (IU/g-pectin), 37 °C and 15 h, respectively. The experiment using optimized parameters in batch fermenter not only resulted in higher ethanol concentration than the one predicted by the model equation, but also saved fermentation time. This study demonstrated that both hydrothermal pretreatment and SSF could be successfully carried out in a single vessel, and use of optimized process parameters helped achieve significant ethanol productivity, indicating commercial potential for the process. To the best of our knowledge, ethanol concentration and ethanol productivity of 28.2 g/l and 2.3 g/l/h, respectively from banana peels have not been reported to date.     

Evaluation of buckwheat and barley tea wastes as ethanol fermentation substrates

Buckwheat tea waste (BWTW) and barley tea waste (BTW), by-products of the beverage industry, are alternative carbohydrate sources for ethanol production. In this study, optimal enzyme loading for enzymatic saccharification of BWTW and BTW was determined, and simultaneous saccharification and fermentation (SSF) was performed by Saccharomyces cerevisiae and Mucor indicus to produce ethanol. Optimal enzyme loading for enzymatic saccharification of 2?% w/v BWTW and BTW was 0.5?% (weight of enzyme/weight of tea wastes) for BWTW and 0.1?% for BTW. Ethanol production from BWTW by S. cerevisiae and M. indicus after 48?h of SSF was 49.9/100?g of BWTW and 47.9/100?g of BWTW, respectively, with 0.5?% enzyme loading. Ethanol production from BTW by S. cerevisiae and M. indicus after 48?h of SSF was 20.5/100?g of BTW and 21.6/100?g of BTW, corresponding to 62 and 66?% of the theoretical yield based on starch content, respectively, with 0.1?% of enzyme loading. Furthermore, S. cerevisiae produced 76?% of the theoretical yield based on the total glucose from starch in BWTW and BTW when a mixture of BWTW and BTW was used as a substrate, with 0.2?% enzyme loading and no additional nitrogen or mineral sources.     

Bioethanol production from enzymatically saccharified lawn clippings from a golf course

The utilization of bioethanol is being focused on as a fuel alternative to oil and or natural gas. Bioethanol production from cellulosic plant residues is one of the solutions proposed for the problems caused by usage of food crops that are also vital for human consumption, such as sugar cane and corn, as a source of bioethanol. However, to utilize these new sources for bioethanol production, conditions for saccharification in each different material have not been optimized. In this study, we reported some optimum conditions for the saccharification of Korean lawn grass (KL) and bent grass (BG) using acremonium cellulase and endoglucanase as saccharifying enzymes for ethanol fermentation. With respect to saccharification of KL and BG, 0.19 and 0.18 g of d-glucose per g-substrate at maximum were produced, respectively. Comminution with a ball mill was found to be effective in the saccharification of KL, while ball-milled BG showed no significant improvement in saccharification. Being incorporated with 99 % of d-glucose consumption, saccharified KL was incubated for 3 days with Saccharomyces cerevisiae and Zymomonas mobilis, respectively, and each mixture fermented to ethanol yielding approximately 100 % of theoretical values from d-glucose consumption, respectively.     

Utilization of Broken Rice for the Production of Poly(3-hydroxybutyrate)

The feasibility of utilizing non edible rice (broken rice) for production of fine materials such as poly(3-hydroxybutyrate) (PHB) was considered as one of the alternative ways of keeping the environment clean for sustainable development. Thus, production of PHB from broken rice by simultaneous saccharification and fermentation (SSF) was investigated. During the SSF process, the rice (15% w/v) material was hydrolyzed to glucose, which was utilized by Cupriavidus necator for growth and production of PHB. The PHB content reached 38% at 58 h fermentation. The PHB had weight average molar mass (Mw) and polydipersity index of 3.82 × 10⁵ (g/mol) and 4.15, respectively. Differential calorimetric scan of the PHB showed a melting temperature (Tm) of 176 °C. Given that the PHB was a homopolymer (which consisted of (R)-3-hydroxybutyric acid monomers), it was thought that broken rice could be a raw material for production of both PHB and (R)-3-hydroxybutyric acid. This SSF process would not only help in the utilization of broken rice or non edible rice, but would also serve as a model for utilization of other raw materials that contain starch for production of PHB.     

Kinetic modeling of enzymatic hydrolysis of pretreated kitchen wastes for enhancing bioethanol production

It is well known that use of low cost and abundant waste materials in microbial fermentations can reduce product costs. Kitchen wastes disposed of in large amounts from cafeterias, restaurants, dining halls, food processing plants, and household kitchens contain high amounts of carbohydrate components such as glucose, starch, and cellulose. Efficient utilization of these sugars is another opportunity to reduce ethanol costs. In this study, the effect of pretreatment methods (hot water, acid solutions, and a control) on enzymatic hydrolysis of kitchen wastes was evaluated using a kinetic modeling approach. Fermentation experiments conducted with and without traditional fermentation nutrients were assessed at constant conditions of pH 4.5 and temperature of 30 °C for 48 h using commercial dry baker’s yeast, Saccharomyces cerevisiae. The control, which involved no treatment, and hot water treated samples gave close glucose concentrations after 6 h. The highest and lowest rates of glucose production were found as 0.644 and 0.128 (h^?1) for the control (or no-pretreated (NPT)) and 1% acid solutions, respectively. The fermentation results indicated that final ethanol concentrations are not significantly improved by adding nutrients (17.2–23.3 g/L). Thus, it was concluded that product cost can be lowered to a large extent if (1) kitchen wastes are used as a substrate, (2) no fermentation nutrient is used, and (3) hydrolysis time is applied for about 6 h. Further optimization study is needed to increase the yield to higher levels.     

Cost-effective approach to ethanol production and optimization by response surface methodology

Food wastes disposed from residential and industrial kitchens have gained attention as a substrate in microbial fermentations to reduce product costs. In this study, the potential of simultaneously hydrolyzing and subsequently fermenting the mixed carbohydrate components of kitchen wastes were assessed and the effects of solid load, inoculum volume of baker’s yeast, and fermentation time on ethanol production were evaluated by response surface methodology (RSM). The enzymatic hydrolysis process was complete within 6 h. Fermentation experiments were conducted at pH 4.5, a temperature of 30 °C, and agitated at 150 rpm without adding the traditional fermentation nutrients. The statistical analysis of the model developed by RSM suggested that linear effects of solid load, inoculum volume, and fermentation time and the quadratic effects of inoculum volume and fermentation time were significant (P < 0.05). The verification experiments indicated that the developed model could be successfully used to predict ethanol concentration at >90% accuracy. An optimum ethanol concentration of 32.2 g/l giving a yield of 0.40 g/g, comparable to yields reported to date, was suggested by the model with 20% solid load, 8.9% inoculum volume, and 58.8 h of fermentation. The results indicated that the production costs can be lowered to a large extent by using kitchen wastes having multiple carbohydrate components and eliminating the use of traditional fermentation nutrients from the recipe.     

Value addition of vegetable wastes by solid-state fermentation using Aspergillus niger for use in aquafeed industry

Vegetable waste typically has high moisture content and high levels of protein, vitamins and minerals. Its value as an agricultural feed can be enhanced through solid-state fermentation (SSF). Two experiments were conducted to evaluate the nutritional status of the products derived by SSF of a mixture of dried vegetable waste powder and oil cake mixture (soybean flour, wheat flour, groundnut oil cake and sesame oil cake at 4:3:2:1 ratio) using fungi Aspergillus niger S₁4, a mangrove isolate, and A. niger NCIM 616. Fermentation was carried out for 9 days at 35% moisture level and neutral pH. Significant (p < 0.05) increase in crude protein and amino acids were obtained in both the trials. The crude fat and crude fibre content showed significant reduction at the end of fermentation. Nitrogen free extract (NFE) showed a gradual decrease during the fermentation process. The results of the study suggest that the fermented product obtained on days 6 and 9 in case of A. niger S₁4 and A. niger NCIM 616 respectively contained the highest levels of crude protein.     

杂色云芝菌产漆酶条件优化及其对碱性紫5BN的降解

通过实验对杂色云芝菌产漆酶的发酵条件进行了优化,结果表明杂色云芝菌产漆酶的最佳发酵条件为:以质量浓度为15 g/L的羧甲基纤维素钠和质量浓度为1.25 g/L的葡萄糖作为复合碳源,以质量浓度为15 g/L的酵母膏作为氮源,在发酵72 h时加入浓度为2mmol/L的2,5-二甲基苯胺,发酵96 h时加入浓度为2 mmol...     

Landfill leachate enhances fermentative hydrogen production from glucose and sugarcane processing derivatives

Fermentation can use renewable raw materials as substrate, which makes it a sustainable method to obtain H₂. This study evaluates H₂ production by a mixed culture from substrates such as glucose and derivatives from sugarcane processing (sucrose, molasses, and vinasse) combined with landfill leachate. The leachate alone was not a suitable substrate for biohydrogen production. However, leachate blended with glucose, sucrose, molasses, or vinasse increased the H₂ production rate by 2.0-, 2.8-, 4.6-, and 0.5-fold, respectively, as compared with the substrates without the leachate. Determination of metals (Cu, Cd, Pb, Hg, Ni, and Fe) at the beginning and at the end of the fermentative assays showed how they were consumed during the fermentation and demonstrated improved H₂ production. During fermentation, Cu, Fe, and Cd were the most consumed leachate metals. The best substrate combination to produce H₂ was molasses and leachate, which gave high volumetric productivity—469 ml H₂/l h. However, addition of the leachate to the substrates stimulated lactic acid formation pathways, which lowered the H₂ yield. The use of leachate combined with sugarcane processing derivatives as substrates could add value to the leachate and reduce its polluting power, generating a clean energy source from renewable raw materials.     

Ethanol production from potato peel waste (PPW)

Considerable concern is caused by the problem of potato peel waste (PPW) to potato industries in Europe. An integrated, environmentally-friendly solution is yet to be found and is currently undergoing investigation. Potato peel is a zero value waste produced by potato processing plants. However, bio-ethanol produced from potato wastes has a large potential market. If Federal Government regulations are adopted in light of the Kyoto agreement, the mandatory blending of bio-ethanol with traditional gasoline in amounts up to 10% will result in a demand for large quantities of bio-ethanol. PPW contain sufficient quantities of starch, cellulose, hemicellulose and fermentable sugars to warrant use as an ethanol feedstock. In the present study, a number of batches of PPW were hydrolyzed with various enzymes and/or acid, and fermented by Saccharomyces cerevisae var. bayanus to determine fermentability and ethanol production. Enzymatic hydrolysis with a combination of three enzymes, released 18.5 g L^?1 reducing sugar and produced 7.6 g L^?1 of ethanol after fermentation. The results demonstrate that PPW, a by-product of the potato industry features a high potential for ethanol production.     

Using the residue of spirit production and bio-ethanol for protein production by yeasts

The residue (vinasse) formed during the distillation of bio-ethanol and cachaça, a traditional rum-type spirit produced from sugar-cane in Brazil, is highly harmful if discharged into the environment due to high values of COD and BOD. One possibility for minimizing the impact of vinasse in soils and waters is to use the residue in the production of microbial biomass for use as an animal feed supplement that will provide high levels on nitrogen (>9% d.m.) and low content of nucleic (?10% d.m.) This paper reports the production and quality of biomass produced from fermentation of Saccharomycescerevisiae and Candidaparapsilosis in culture media under 12 different culture conditions and the respective effects of each variable (glucose, yeast extract, peptone, potassium phosphate, vinasse, pH and temperature). Of the S. cerevisiae isolates tested, two (VR1 and PE2) originating from fuel alcohol-producing plants were identified as offering the best potential for the industrial production of single cell protein from vinasse due to highest biomass productivity. Our results showed a potential viable and economic use of vinasse.     

Effects of washing with water on enzymatic saccharification and d-lactic acid production from steam-exploded sugarcane bagasse

This study investigated the production of d-lactic acid from unutilized sugarcane bagasse using steam explosion pretreatment. The optimal steam pressure for a steaming time of 5?min was determined. The steam-exploded sugarcane bagasse was hydrolyzed using cellulase (Meicelase) and then the hydrolyzate was subjected to fermentation substrate. By enzymatic saccharification using Meicelase, the highest recovery of glucose from raw bagasse, 73.7?%, was obtained at a steam pressure of 20?atm. For extracted residue with water after steam explosion, the glucose recovery increased up to 94.9?% at a steam pressure of 20?atm. These results showed that washing with water is effective in removing enzymatic reaction inhibitors. After steam pretreatment (steam pressure of 20?atm), d-lactic acid was produced by Lactobacillus delbrueckii NBRC 3534 from the enzymatic hydrolyzate of steam-exploded bagasse and washed residue. The conversion rate of d-lactic acid obtained from the glucose concentration was 66.6?% for the hydrolyzate of steam-exploded bagasse without washing with water and 90.0?% for that derived from the extracted residue with water after steam explosion. These results also demonstrated that the hydrolyzate of steam-exploded bagasse (without washing with water) contains fermentation inhibitors and washing with water can remove them.     

Physicochemical and biochemical changes during composting of different mixing ratios of biogas sludge with palm oil mill wastes and biogas effluent

Prior to composting, the composition of palm oil mill wastes were analyzed. Palm empty fruit bunches (PEFB) contained the highest total organic carbon (52.83 % dry weight) while palm oil mill biogas sludge (POMS) and decanter cake (DC) contained higher total nitrogen (3.6 and 2.37 % dry weight, respectively) than the others. In addition, palm oil fuel ash (POFA) had a high amount of phosphorus and potassium (2.17 and 1.93 % dry weight, respectively). The effect of mixture ratio of POMS and other palm oil mill wastes for composting was studied using the mixed culture Super LDD1 as an inoculum. All compost piles turned dark brown and attained an ambient temperature after 40 days incubation. The pH values were stable in the range of 6.9–7.8 throughout the process whereas the moisture content tended to decrease till the end with the final value around 30 %. After 60 days incubation, the mixture ratio of POMS:PEFB:DC at 2:1:1 with the addition of biogas effluent gave the highest quality of the compost. Its nitrogen content was 31.75 % higher than the other treatments that may be a result of growth of ink cap mushroom (Coprinus sp.). This is the first report on the occurrence of this mushroom during composting. In addition, its nutrients (3.26 % N, 0.84 % P and 2.03 % K) were higher than the level of the Organic Fertilizer Standard. The mixed culture Super LDD1 produced the highest activity of CMCase (6.18 Unit/g) and xylanase (11.68 Unit/g) at 9 days fermentation. Therefore, this solid-state fermentation could be employed for production of compost as well as enzymes.     

Groundwater and Nutrient Discharge into Jiaozhou Bay,North China

The health of near shore marine ecosystems has long been a concern because of its importance to coastal areas. Jiaozhou Bay (JZB) is one such marine ecosystem experiencing rapid water quality degradation in the last several decades. From the area surrounding the bay, the nutrients discharged into the bay through surface water and groundwater has been greatly changed. The thickness of the aquifers and the permeability is relatively high, the concentrations of nutrients in the groundwater are generally high, and so the groundwater discharged into JZB is very significant. However, no attempt has ever been made to evaluate the amount of nutrients discharged into the bay area via groundwater. In this study, the cross-section method and water balance method were used to estimate the amount of groundwater and nutrients discharged into JZB via the subsurface. Groundwater was monitored and sampled at aquifers surrounding the bay area, and some previously available data was also analyzed. The results indicated that groundwater from the Baisha Aquifer east of JZB now is the major source of nutrients (nitrate, dissolved SiO₂) being discharged into the bay. The concentrations of nutrients in the groundwater have been increasing with intensive agricultural land use. However, Dagu Aquifer, the largest aquifer north of JZB, only provides limited nutrients to the bay area because of the construction of a low permeability subsurface dam. Historically, during the 1970s to the 1990s, the Baisha Aquifer experienced seawater intrusion due to excessive groundwater withdrawal. The same was true for the Dagu Aquifer from the 1980s to the 1990s. Because of this, no significant nutrients were discharged into the bay.     

Conversion of CCA-treated wood to ethanol: a method to reduce leaching of metals from disposed treated wood prior to disposal

The objective of this paper is to evaluate the feasibility of producing ethanol from CCA-treated wood that is highly leachable. Following the initial tests, CCA-treated wood was hydrolysed and fermented and the results showed not only that ethanol was produced during the fermentation process but that metals were taken up by the yeast. Toxicity characteristic leaching procedure tests of the hydrolysed wood leached less than 4 mg/L of As while minimal amounts of Cr and Cu remained in the hydrolysed wood which makes landfilling of hydrolysed wood acceptable and less hazardous. A slightly lower amount of ethanol from CCA-treated than untreated wood was produced (6 and 7 g/L, respectively). In general, it suggests that production of ethanol as a source of energy from a hazardous waste (CCA-treated wood) is feasible.     

An international study: Effect of farm manure on the release of phosphorus from fly ash

Fly ash is abundantly produced from thermal power plants and is considered a hazardous waste. However, in recent years, fly ash has been widely utilized in the agricultural sector as a soil modifier. It is particularly important for wasteland/mine spoil reclamation due to its ability to provide a source of plant nutrients and improve physicochemical properties of soil. Although fly ash itself contains many plant nutrients, most nutrients, including phosphorus (P), are in a bound form not easily available to plants. This study analyzed the effect of farm manure on the solubility of P from fly ash. Incubation studies were conducted to determine the effect of farm manure on P solubilization to use as a potential option for remediation. © 2007 Wiley Periodicals, Inc.     

有机固体废物生物法制氢的研究进展

综述了利用有机固体废物生物法制氢的原理和研究现状。城市有机固体废物、农业有机废物、工业有机废物是生物法制氢的主要原料。暗发酵制氢是利用有机废物厌氧消化的产酸阶段而产氢，pH、温度、水力停留时间、氢气分压、原料性质、微量元素含量、产甲烷微生物抑制剂等均影响氢气产率。光发酵制氢是利用光合厌氧细菌将挥发性有机酸转化为氢气和二氧化碳。暗发酵和光发酵制氢时，生物固定化有利于高速连续产氢。在有机废物处理和生物法制氢方面，暗发酵一光发酵、暗发酵一微生物燃料电池的组合工艺是具有前景的技术。今后的研究方向是原料的预处理技术、选育高效产氢菌株、发明高效反应器、优化处理工艺和处理条件等。     

A novel process for ethanol or biogas production from cellulose in blended-fibers waste textiles

A novel process has been developed for separation of the cellulose, i.e. cotton and viscose, from blended-fibers waste textiles. An environmentally friendly cellulose solvent, N-methylmorpholine-N-oxide (NMMO) was used in this process for separation and pretreatment of the cellulose. This solvent was mixed with blended-fibers textiles at 120 °C and atmospheric pressure to dissolve the cellulose and separate it from the undissolved non-cellulosic fibers. Water was then added to the solution in order to precipitate the cellulose, while both water and NMMO were reused after separation by evaporation. The cellulose was then either hydrolyzed by cellulase enzymes followed by fermentation to ethanol, or digested directly to produce biogas. The process was verified by testing 50/50 polyester/cotton and 40/60 polyester/viscose-blended textiles. The polyesters were purified as fibers after the NMMO treatments, and up to 95% of the cellulose fibers were regenerated and collected on a filter. A 2-day enzymatic hydrolysis and 1-day fermentation of the regenerated cotton and viscose resulted in 48 and 50 g ethanol/g regenerated cellulose, which were 85% and 89% of the theoretical yields, respectively. This process also resulted in a significant increase of the biogas production rate. While untreated cotton and viscose fibers were converted to methane by respectively, 0.02% and 1.91% of their theoretical yields in 3 days of digestion, the identical NMMO-treated fibers resulted into about 30% of yield at the same period of time.