Anaerobic Digestion of Sugarcane Vinasse Through a Methanogenic UASB Reactor Followed by a Packed Bed Reactor

The anaerobic treatment of raw vinasse in a combined system consisting in two methanogenic reactors, up-flow anaerobic sludge blanket (UASB) + anaerobic packed bed reactors (APBR), was evaluated. The organic loading rate (OLR) was varied, and the best condition for the combined system was 12.5 kg COD m^?3day^?1 with averages of 0.289 m³ CH₄ kg COD r^?1for the UASB reactor and 4.4 kg COD m^?3day^?1 with 0.207 m³ CH₄ kg COD r^?1 for APBR. The OLR played a major role in the emission of H₂S conducting to relatively stable quality of biogas emitted from the APBR, with H₂S concentrations <10 mg L^?1. The importance of the sulphate to COD ratio was demonstrated as a result of the low biogas quality recorded at the lowest ratio. It was possible to develop a proper anaerobic digestion of raw vinasse through the combined system with COD removal efficiency of 86.7% and higher CH₄ and a lower H₂S content in biogas.     

Viability of Using Glycerin as a Co-substrate in Anaerobic Digestion of Sugarcane Stillage (Vinasse): Effect of Diversified Operational Strategies

Vinasse, from sugar and ethanol production, stands out as one of the most problematic agroindustry wastes due to its high chemical oxygen demand, large production volume, and recalcitrant compounds. Therefore, the viability of using glycerin as a co-substrate in vinasse anaerobic digestion was tested, to increase process efficiency and biogas productivity. The effect of feeding strategy, influent concentration, cycle length, and temperature were assessed to optimize methane production. Glycerin (1.53% v/v) proved to be a good co-substrate since it increased the overall methane production in co-digestion assays. CH₄ productivity enhanced exponentially as influent concentration increased, but when temperature was increased to 35 °C, biogas production was impaired. The highest methane productivity and yield were achieved using fed-batch mode, at 30 °C and at an organic loading rate of 10.1 kg COD m⁻³ day⁻¹: 139.32 mol CH₄ m⁻³ day⁻¹, 13.86 mol CH₄ kg COD_applied, and 15.30 mol CH₄ kg COD_removed. Methane was predominantly produced through the hydrogenotrophic route. In order to treat all the vinasse produced by a mid-size sugar and ethanol plant, nine reactors with 7263.4 m³ each would be needed. The energy generated by burning the biogas in boilers would reach approximately 92,000 MW h per season and could save up to US$ 240,000.00 per month in diesel oil demand.

     

Anaerobic Digestion of Yard Waste with Hydrothermal Pretreatment

The digestibility of lignocellulosic biomass is limited by its high content of refractory components. The objective of this study is to investigate hydrothermal pretreatment and its effects on anaerobic digestion of sorted organic waste with submerged fermentation. Hydrothermal pretreatment (HT) was performed prior to anaerobic digestion, and three agents were examined for the HT: hot compressed water, alkaline solution, and acidic solution. The concentrations of glucose and xylose were the highest in the sample pretreated in acidic solution. Compared with that of the untreated sample, the biogas yields from digesting the samples pretreated in alkaline solution, acidic solution, and hot water increased by 364, 107, and 79 %, respectively. The decrease of chemical oxygen demand (COD) in liquid phase followed the same order as for the biogas yield. The initial ammonia content of the treated samples followed the order sample treated in acidic solution > sample treated in alkaline solution > sample treated in hot water. The concentrations of volatile fatty acids (VFAs) were low, indicating that the anaerobic digestion process was running at continuously stable conditions.     

Biogas Plasticization Coupled Anaerobic Digestion: The Anaerobic Pump Stoichiometry

This paper presents the stoichiometry section of a bioenergetics investigation into the biogas plasticization of wastewater sludge using the Anaerobic Pump (TAP). Three residue samples, an input substrate and two residual products, were collected from two side by side operated AD systems, a conventional continuous flow and stirred reactor, and TAP, and submitted for elemental and calorimetric analyses. The elemental compositions of the residues were fitted to a heterotrophic metabolism model [1] for both systems. To facilitate balanced stoichiometric models, a simple “cell” correction computation separates measured residual composites into “real” residual composition and cell growth (C₅H₇NO₂) components. The elemental data and model results show that the TAP stage II residual composition (C₁H_0.065O_0.0027N_0.036) was nearly devoid of hydrogen and oxygen, leaving only fixed carbon and cells grown as the composition of the remaining mass. This quantitative evidence supports prior measurements of very high methane yields from TAP stage II reactor during steady-state experiments [2]. All performance parameters derived from the stoichiometric model(s) showed good agreement with measured steady-state averaged values. These findings are strong evidence that plasticization–disruption (TAP) cycle is the mechanism responsible for the observed increases in methane yield. The accuracy achieved by the stoichiometry models qualifies them for thermodynamic analysis to obtain potentials and bioconversion efficiencies. How applied pressure causes matrix conformation changes triggered by a functional consequence (plasticization and disruption) is this study’s essential focus.     

Soaking Pretreatment of Corn Stover for Bioethanol Production Followed by Anaerobic Digestion Process

The production of ethanol and methane from corn stover (CS) was investigated in a biorefinery process. Initially, a novel soaking pretreatment (NaOH and aqueous-ammonia) for CS was developed to remove lignin, swell the biomass, and improve enzymatic digestibility. Based on the sugar yield during enzymatic hydrolysis, the optimal pretreatment conditions were 1?% NaOH?+?8?% NH₄OH, 50°C, 48?h, with a solid-to-liquid ratio 1:10. The results demonstrated that soaking pretreatment removed 63.6?% lignin while reserving most of the carbohydrates. After enzymatic hydrolysis, the yields of glucose and xylose were 78.5?% and 69.3?%, respectively. The simultaneous saccharification and fermentation of pretreated CS using Pichia stipitis resulted in an ethanol concentration of 36.1?g/L, corresponding only to 63.3?% of the theoretical maximum. In order to simplify the process and reduce the capital cost, the liquid fraction of the pretreatment was used to re-soak new CS. For methane production, the re-soaked CS and the residues of SSF were anaerobically digested for 120?days. Fifteen grams CS were converted to 1.9?g of ethanol and 1337.3?mL of methane in the entire process.     

Biogas Plasticization Coupled Anaerobic Digestion: Anaerobic Pump Calorimetry

Applied Biochemistry and Biotechnology - This paper presents the quantitative bomb calorimetric high heat values (HHV) for residue samples collected from the Anaerobic Pump (®TAP) and a...     

Antioxidant Activity and Kinetic Characterization of Chlorella vulgaris Growth under Flask-Level Photoheterotrophic Growth Conditions

C. vulgaris is a unicellular microalgae, whose growth depends on the conditions in which it is found, synthesizing primary and secondary metabolites in different proportions. Therefore, we analyzed and established conditions in which it was possible to increase the yields of metabolites obtained at the flask level, which could then be scaled to the photobioreactor level. As a methodology, a screening design was applied, which evaluated three factors: type of substrate (sodium acetate or glycerol); substrate concentration; and exposure-time to red light (photoperiod: 16:8 and 8:16 light/darkness). The response variables were: cell division; biomass; substrate consumption; and antioxidant activity in intracellular metabolites (ABTS•+ and DPPH•). As a result, the sodium acetate condition of 0.001 g/L, in a photoperiod of 16 h of light, presented a doubling time (Td = 4.84 h) and a higher rate of division (σ = 0.20 h⁻¹), having a final biomass concentration of 2.075 g/L. In addition, a higher concentration of metabolites with antioxidant activity was found in the sodium acetate (0.629 Trolox equivalents mg/L ABTS•+ and 0.630 Trolox equivalents mg/L DPPH•). For the glycerol, after the same photoperiod (16 h of light and 8 h of darkness), the doubling time (Td) was 4.63 h, with a maximum division rate of σ = 0.18 h⁻¹ and with a biomass concentration at the end of the kinetics of 1.4 g/L. Sodium acetate under long photoperiods, therefore, is ideal for the growth of C. vulgaris, which can then be scaled to the photobioreactor level.     

小球藻对碘富集实验的研究

用碘离子电极法对三种制约小球藻富集碘量的因素进行了研究。结果表明,这些因素对小球藻富集碘都有影响。当KI质量浓度为800μg/mL,pH值为7.0,为它们各自作为单一制约因素时的最佳条件,此时小球藻对碘的富集效果最佳,试验中也看到小球藻在这些条件下生长处于最佳状态。     

Biogas Plasticization Coupled Anaerobic Digestion: Continuous Flow Anaerobic Pump Test Results

In this investigation, the Anaerobic Pump (®TAP) and a conventional continuous flow stirred tank reactor (CFSTR) were tested side by side to compare performance. TAP integrates anaerobic digestion (AD) with biogas plasticization–disruption cycle to improve mass conversion to methane. Both prototypes were fed a “real world” 50:50 mixture of waste-activated sludge (WAS) and primary sludge and operated at room temperature (20°C). The quantitative results from three steady states show TAP peaked at 97% conversion of the particulate COD in a system hydraulic residence time (HRT) of only 6 days. It achieved a methane production of 0.32 STP cubic meter CH₄ per kilogram COD fed and specific methane yield of 0.78 m³ CH₄ per cubic meter per day. This was more than three times the CFSTR specific methane yield (0.22 m³ CH₄ per cubic meter per day) and more than double the CFSTR methane production (0.15 m³ CH₄ per kilogram COD fed). A comparative kinetics analysis showed the TAP peak substrate COD removal rate (R _o) was 2.24 kg COD per cubic meter per day, more than three times the CFSTR substrate removal rate of 0.67 kg COD per cubic meter per day. The three important factors contributing to the superior TAP performance were (1) effective solids capture (96%) with (2) mass recycle and (3) stage II plasticization–disruption during active AD. The Anaerobic Pump (®TAP) is a high rate, high efficiency–low temperature microbial energy engine that could be used to improve renewable energy yields from classic AD waste substrates like refuse-derived fuels, treatment plant sludges, food wastes, livestock residues, green wastes and crop residuals.     

Microcalorimetric Monitoring of Anaerobic Digestion Processes

Microcalorimetry was used for monitoring anaerobic digestion processes of heavily polluted industrial waste waters (from cheese industry, distilleries, yeast plant). Interpreting the thermal power-time curves by HPLC, some sub-processes in batch cultures were tentatively identified as acidogenic, acetogenic and methanogenic. Processes underlying power-time curves up to 10 h were different for different wastes. In the case of cheese whey and distillery waste it was acidogenesis, in the case of sulfate containing waste - presumably reduction of sulfates. The effect of Biotreat 100 (BimKemi Eesti Ltd.), a preparation for removing H₂S from waste water, was observed for these processes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Combined Mesophilic Anaerobic and Thermophilic Aerobic Digestion Process: Effect on Sludge Degradation and Variation of Sludge Property

One-stage autothermal thermophilic aerobic digestion (ATAD) is effective for the reduction of volatile solids (VSs) and pathogen in sewage sludges. A novel process of combining mesophilic (<35 °C) anaerobic digestion with a thermophilic (55 °C) aerobic digestion process (AN/TAD) occurred in a one-stage digester, which was designed for aeration energy savings. The efficiency of sludge degradation and variation of sludge properties by batch experiments were evaluated for the AN/TAD digester with an effective volume of 23 L for 30 days compared with conventional thermophilic aerobic digestion (TAD). The AN/TAD system can efficiently achieve sludge stabilization on the 16th day with a VS removal rate of 38.1 %. The AN/TAD system was operated at lower ORP values in a digestion period with higher contents of total organic compounds, volatile fatty acids, protein, and polysaccharide in the soluble phase than those of the TAD system, which can rapidly decreased and had low values in the late period of digestion for the AN/TAD system. In the AN/TAD system, intracellular substances had lysis because of initial hydrolytic acidification.     

Composition of the neutral lipids of Chlorella vulgaris

Summary The qualitative and quantitative compositions of the neutral lipids and the fatty-acid composition of the individual classes of neutral lipids isolated from fresh, freeze-dried, and air-dry cells of the algaChlorella vulgaris have been investigated.The presence of natural fatty acid methyl esters in the lipid extract of the fresh and air-dry chlorella cells has been established.It has been shown that the fatty acid methyl ester fraction is enriched with palmitic acid as compared with the triglyceride and free fatty acid fractions.The main unsaturated acids of the chlorella lipids contain the first double bond in the ⁹ position.Division of Microbiology of the Academy of Sciences of the Uzbek SSR, Tashkent. Institute of the Chemistry of Plant Substances, Academy of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 49–53, January–February, 1978.     

Lime Pretreatment of Sugarcane Bagasse for Bioethanol Production

The pretreatment of sugarcane bagasse with lime (calcium hydroxide) is evaluated. The effect of lime pretreatment on digestibility was studied through analyses using central composite design (response surface), considering pretreatment time, temperature, and lime loading as factors. The responses evaluated were the yield of glucose from pretreated bagasse after enzymatic hydrolysis. Experiments were performed using the bagasse as it comes from an alcohol/sugar factory (non-screened bagasse) and bagasse in the size range from 0.248 to 1.397 mm (screened bagasse) (12-60 mesh). It was observed that the particle size presented influence in the release of fermentable sugars after enzymatic hydrolysis using low loading of cellulase and β-glucosidase (3.5 FPU/g dry pretreated biomass and 1.0 IU/g dry pretreated biomass, respectively).     

Biotransformation of arsenate to arsenosugars by Chlorella vulgaris

Chlorella vulgaris was cultivated in a growth medium containing arsenate concentration of <0.01, 10, 100 and 1000 mg l^?1. Illumination was carried out in 12 h cycles for 5 days. The health status of the culture was monitored by continuous pH and dissolved oxygen (DO) readings. Destructive sampling was used for the determination of biomass, chlorophyll, total arsenic and arsenic species. The chlorophyll a content, the DO and pH cycles were not significantly different for the different arsenate concentrations in the culture. In contrast, biomass production was significantly (p < 0.05) increased for the arsenic(V) treatment at 1000 mg l^?1 compared with 100 mg l^?1. The arsenic concentration in the algae increased with the arsenate concentration in the culture. However, the bioconcentration factor decreased a hundred‐fold with increase of arsenate from the background level to 1000 mg l^?1. The arsenic species were identified by using strong anion‐exchange high‐performance liquid chromatography–inductively coupled plasma mass spectrometry analysis after methanol/water (1 : 1) extraction. The majority (87–100%) of the extractable arsenic was still arsenate; arsenite was found to be between 1 and 6% of total extractable arsenic in the algae. In addition to dimethylarsinic acid, one unknown arsenical (almost co‐eluting with methylarsonic acid) and three different arsenosugars have been identified for the first time in C. vulgaris growing in a culture containing a mixture of antibiotics and believed to be axenic. The transformation to arsenosugars in the algae is not dependent on the arsenate concentration in the culture and varies between 0.2 and 5% of total accumulated arsenic. Although no microbiological tests for bacterial contamination were made, this study supports the hypothesis that algae, and not associated bacteria, produce the arsenosugars. Copyright © 2003 John Wiley & Sons, Ltd.     

Anaerobic Digestion of Cassava Wastewater Pre-treated by Fungi

Cassava wastewater (cww) contains high concentrations of easily acidifying compounds, requiring a buffered system to allow a stable operation during anaerobic digestion (AD). The possibility to include a preliminary one-step fungi treatment aimed at raising the pH and buffering the cww prior to AD was studied. Preliminary tests were performed with a naturally grown fungal mixed culture, under aerated (AE), non-aerated (NAE) and initially oxygen-deprived (IOD) conditions. The cww was pre-treated by the NAE condition, until reaching a soluble chemical oxygen demand (COD) of 10 g?L^?1 and pH 6.4 (batch A) and pH 5.7 (batch B). The fungal mixed culture showed ability to biodegrade the cww with initial pH of 4.4 and 14,500 mg?COD?L^-1, raising the pH over 8.5, with only 13 % of COD remaining within 27 days for both AE and NAE condition. The fungal pre-treated-cww (FPTcww) was subjected to anaerobic digestion under different buffered (CaCO₃ and NaHCO₃) and non-buffered conditions. The FPTcww with initial pH at 6.4 provided stability during the anaerobic biodegradability tests, showing the possibility of system operation without buffer addition, with final pH around 7. The application of a fungal pre-treatment can be a promising strategy to permit the anaerobic digestion of carbohydrate-rich wastewaters.     

厌氧消化过程化学变化的教学研究

有氧和无氧分解是生物体代谢有机物的2种途径,前者一直是生物化学课程中物质代谢的教学核心和重点。厌氧消化是有机物经无氧分解生成甲烷和二氧化碳的途径,相比于有氧分解,其具有化学反应众多、中间产物多、途径复杂等特点,同时对维护自然生态平衡、物质循环和人类社会的绿色健康发展等具有重要作用。如此重要的途径长期以来一直没有在生物化学教材与教学内容中体现,亟需进行教学改革。利用布鲁姆教学目标分类理论将厌氧消化过程中的主要反应、氧化还原过程及电子传递、热力学变化及串联反应等知识进行系统介绍,对物质代谢的另一条无氧路径进行补充,丰富了学生代谢网络的知识框架,完善了生物化学教学体系,相关教学内容和经验可供同行参考借鉴,对相关课程教学改革具有重要价值。     

The association mode of arsenic accumulated in the freshwater alga Chlorella vulgaris

Arsenic accumulated in living Chlorella vulgaris cells was solvent-fractionated with chloroform/methanol (2:1), and the fractions were analyzed for arsenic. A large part of the accumulated arsenic was localized in the extract residues. The extract residue from the same extraction of C. vulgaris, which had been, however, cultured in any arsenic-free Detmer medium (MD), adsorbed arsenic physico-chemically at a concentration of 1.1 mg As g^?1 dry weight. Arsenic was found to be combined with protein with molecular weight around 3000 in the arsenicaccumulated living cells. The arsenic-bound protein was analyzed for amino acids. The experimental results showed that no metallothionein-like protein was inductively biosynthesized in C. vulgaris on the exposure to arsenic.     

Bioactive Hydrolysates from Chlorella vulgaris: Optimal Process and Bioactive Properties

Microalgae have been described as a source of bioactive compounds, such as peptides. Microalgae are easy to produce, making them a sustainable resource for extracting active ingredients for industrial applications. Several microalgae species have interesting protein content, such as Chlorella vulgaris with around 52.2% of protein, making it promising for peptide hydrolysate production. Therefore, this work focused on the production of water-soluble hydrolysates rich in proteins/peptides from the microalgae C. vulgaris and studied bioactive properties. For that, a design of experiments (DOE) was performed to establish the optimal conditions to produce hydrolysates with higher levels of protein, as well as antioxidant and antihypertensive properties. Four experimental factors were considered (cellulase percentage, protease percentage, hydrolysis temperature, and hydrolysis duration) for three responses (protein content, antioxidant activity, and antihypertensive activity). The optimal conditions determined by the DOE allowed producing a scaled-up hydrolysate with 45% protein, with antioxidant activity, measured by oxygen radical absorbance capacity assay, of 1035 µmol TE/g protein, IC₅₀ for angiotensin-converting enzyme inhibition activity of 286 µg protein/mL, and α-glucosidase inhibition of 31% (30 mg hydrolysate/mL). The obtained hydrolysates can be used as functional ingredients for food and nutraceuticals due to their antioxidant, antihypertensive, and antidiabetic potential. Moreover, the antioxidant potential of the extracts may be relevant for the cosmetic industry, especially in antiaging formulations.     

Atmospheric Pressure Plasma Pretreatment of Sugarcane Bagasse: The Influence of Moisture in the Ozonation Process

Sugarcane bagasse samples were pretreated with ozone via atmospheric O₂ pressure plasma. A delignification efficiency of approximately 80 % was observed within 6 h of treatment. Some hemicelluloses were removed, and the cellulose was not affected by ozonolysis. The quantity of moisture in the bagasse had a large influence on delignification and saccharification after ozonation pretreatment of the bagasse, where 50 % moisture content was found to be best for delignification (65 % of the cellulose was converted into glucose). Optical absorption spectroscopy was applied to determine ozone concentrations in real time. The ozone consumption as a function of the delignification process revealed two main reaction phases, as the ozone molecules cleave the strong carbon–carbon bonds of aromatic rings more slowly than the weak carbon–carbon bonds of aliphatic chains.