Microbial communities involved in biogas production from wheat straw as the sole substrate within a two-phase solid-state anaerobic digestion

Microbial communities involved in biogas production from wheat straw as the sole substrate were investigated. Anaerobic digestion was carried out within an up-flow anaerobic solid-state (UASS) reactor connected to an anaerobic filter (AF) by liquor recirculation. Two lab-scale reactor systems were operated simultaneously at 37 °C and 55 °C. The UASS reactors were fed at a fixed organic loading rate of 2.5 g L⁻¹ d⁻¹, based on volatile solids. Molecular genetic analyses of the bacterial and archaeal communities within the UASS reactors (digestate and effluent liquor) and the AFs (biofilm carrier and effluent liquor) were conducted under steady-state conditions. The thermophilic UASS reactor had a considerably higher biogas and methane yield in comparison to the mesophilic UASS, while the mesophilic AF was slightly more productive than the thermophilic AF. When the thermophilic and mesophilic community structures were compared, the thermophilic system was characterized by a higher Firmicutes to Bacteroidetes ratio, as revealed by 16S rRNA gene (rrs) sequence analysis. The composition of the archaeal communities was phase-separated under thermophilic conditions, but rather stage-specific under mesophilic conditions. Family- and order-specific real-time PCR of methanogenic Archaea supported the taxonomic distribution obtained by rrs sequence analysis. The higher anaerobic digestion efficiency of the thermophilic compared to the mesophilic UASS reactor was accompanied by a high abundance of Firmicutes and Methanosarcina sp. in the thermophilic UASS biofilm.     

Effect of reactor configuration on biogas production from wheat straw hydrolysate

The potential of wheat straw hydrolysate for biogas production was investigated in continuous stirred tank reactor (CSTR) and up-flow anaerobic sludge bed (UASB) reactors. The hydrolysate originated as a side stream from a pilot plant pretreating wheat straw hydrothermally (195 °C for 10–12 min) for producing 2nd generation bioethanol [Kaparaju, P., Serrano, M., Thomsen, A.B., Kongjan, P., Angelidaki, I., 2009. Bioethanol, biohydrogen and biogas production from wheat straw in a biorefinery concept. Bioresource Technology 100 (9), 2562–2568]. Results from batch assays showed that hydrolysate had a methane potential of 384 ml/g-volatile solids (VS)_added. Process performance in CTSR and UASB reactors was investigated by varying hydrolysate concentration and/or organic loading rate (OLR). In CSTR, methane yields increased with increase in hydrolysate concentration and maximum yield of 297 ml/g-COD was obtained at an OLR of 1.9 g-COD/l d and 100% (v/v) hydrolysate. On the other hand, process performance and methane yields in UASB were affected by OLR and/or substrate concentration. Maximum methane yields of 267 ml/g-COD (COD removal of 72%) was obtained in UASB reactor when operated at an OLR of 2.8 g-COD/l d but with only 10% (v/v) hydrolysate. However, co-digestion of hydrolysate with pig manure (1:3 v/v ratio) improved the process performance and resulted in methane yield of 219 ml/g-COD (COD removal of 72%). Thus, anaerobic digestion of hydrolysate for biogas production was feasible in both CSTR and UASB reactor types. However, biogas process was affected by the reactor type and operating conditions.     

Biomethanation of rice and wheat straw

When rice or wheat straw was added to cattle dung slurry and digested anaerobically, daily gas production increased from 176 to 331 l/kg total solids with 100% rice straw and to 194 l/kg total solids with 40% wheat straw. Not only was methane production enhanced by adding chopped crop residues but a greater biodegradability of organic matter in the straws was achieved.The authors are with the Microbiology and Molecular Genetics Unit, Tata Energy Research Institute, 158 Jor Bagh, New Delhi 110 003, India     

In vitro degradation of wheat straw by anaerobic fungi from small ruminants

Abstract

Anaerobic ruminal fungi may play an active role in fibre degradation as evidenced by the production of different fibrolytic enzymes in culture filtrate. In the present study, 16 anaerobic fungal strains were isolated from ruminal and faecal samples of sheep and goats. Based on their morphological characteristics they were identified as species of Anaeromyces, Orpinomyces, Piromyces and Neocallimastix. Isolated Neocallimastix sp. from goat rumen showed a maximum activity of CMCase (47.9 mIU ml^?1) and filter paper cellulase (48.3 mIU ml^?1), while Anaeromyces sp. from sheep rumen showed a maximum xylanolytic activity (48.3 mIU ml^?1). The cellobiase activity for all the isolates ranged from 178.0 – 182.7 mIU ml^?1. Based on the enzymatic activities, isolated Anaeromyces sp. from sheep rumen and Neocallimastix sp. from goat rumen were selected for their potential of in vitro fibre degradation. The highest in vitro digestibility of NDF (23.2%) and DM (34.4%) was shown for Neocallimastix sp. from goat rumen, as compared to the digestibility of NDF and DM in the control group of 17.5 and 25.0%, respectively.     

Industrial by-products from white-rot fungi production. Part II: Application in anaerobic digestion for enzymatic treatment of hay and straw

By-products of white-rot fungi cultivations are valuable resources for the production of useful enzyme cocktails. These enzymes, which act synergistically to deconstruct lignocellulose polymers, can be recovered and potentially applied in industrial processes. This study investigated the application of processed by-products from Lentinula edodes cultivations in mesophilic and thermophilic anaerobic digestions of hay and straw. Untreated and mechanically treated hay and straw were investigated in biochemical methane potential assays with or without application of enzyme-containing materials. Biomasses, inocula and processed by-product were analyzed chemically and the degradation rate of lignocellulose polymers determined.In mesophilic conditions, all of the fungus-derived enzyme treatments increased the methane yield. A newly generated enzyme preparation significantly enhanced the methane yield of chopped hay and straw, and accelerated the rate of hemicellulose degradation. In general, the degree of cellulose degradation correlated with the methane yield. The novel enzyme preparation contains a larger variety of enzymes than is commonly found in biogas enzyme preparations and is thus an attractive candidate for significant process improvement. Our new investigation further shows that enzyme preparations of L. edodes have a high potential for catalytic activity in lignocellulose-rich systems without manure as co-substrate.     

The preparation of high-grade bio-oils through the controlled, low temperature microwave activation of wheat straw

The low temperature microwave activation of biomass has been investigated as a novel, energy efficient route to bio-oils. The properties of the bio-oil produced were considered in terms of fuel suitability. Water content, elemental composition and calorific value have all been found to be comparable to and in many cases better than conventional pyrolysis oils. Compositional analysis shows further differences with conventional pyrolysis oils including simpler chemical mixtures, which have potential as fuel and chemical intermediates. The use of simple additives, e.g. HCl, H₂SO₄ and NH₃, affects the process product distribution, along with changes in the chemical composition of the oils. Clearly the use of our low temperature technology gives significant advantages in terms of preparing a product that is much closer to that which is required for transport fuel applications.     

The future of anaerobic digestion and biogas utilization

One of the common tendencies of animal production activities in Europe and in developed countries in general is to intensify the animal production and to increase the size of the animal production units. High livestock density is always accompanied by production of a surplus of animal manure, representing a considerable pollution threat for the environment in these areas. Avoiding over-fertilization is not only important for environmental protection reasons but also for economical reasons. Intensive animal production areas need therefore suitable manure management, aiming to export and to redistribute the excess of nutrients from manure and to optimize their recycling.     

Characteristics of degraded cellulose obtained from steam-exploded wheat straw

The isolation of cellulose from wheat straw was studied using a two-stage process based on steam explosion pre-treatment followed by alkaline peroxide post-treatment. Straw was steamed at 200 degrees C, 15 bar for 10 and 33 min, and 220 degrees C, 22 bar for 3, 5 and 8 min with a solid to liquid ratio of 2:1 (w/w) and 220 degrees C, 22 bar for 5 min with a solid to liquid ratio of 10:1, respectively. The steamed straw was washed with hot water to yield a solution rich in hemicelluloses-derived mono- and oligosaccharides and gave 61.3%, 60.2%, 66.2%, 63.1%, 60.3% and 61.3% of the straw residue, respectively. The washed fibre was delignified and bleached by 2% H2O2 at 50 degrees C for 5 h under pH 11.5, which yielded 34.9%, 32.6%, 40.0%, 36.9%, 30.9% and 36.1% (% dry wheat straw) of the cellulose preparation, respectively. The optimum cellulose yield (40.0%) was obtained when the steam explosion pre-treatment was performed at 220 degrees C, 22 bar for 3 min with a solid to liquid ratio of 2:1, in which the cellulose fraction obtained had a viscosity average degree of polymerisation of 587 and contained 14.6% hemicelluloses and 1.2% klason lignin. The steam explosion pre-treatment led to a significant loss in hemicelluloses and alkaline peroxide post-treatment resulted in substantial dissolution of lignin and an increase in cellulose crystallinity. The six isolated cellulose samples were further characterised by FT-IR and 13C-CP/MAS NMR spectroscopy and thermal analysis.     

Studies on biological decomposition of wheat straw

Summary The incorporation of undecomposed wheat straw in the soil along-with the micro-organisms favourably increased the yield of groundnut crop. An increase of 37 per cent in yield was recorded when wheat straw was inoculated withPenicillium digitatum and the C:P ratio was adjusted to 65. Inoculated treatments of narrower C:P ratio gave a higher yield than wider C:P ratio treatments inoculated with the same cultures. An increase in nitrogen uptake by groundnut plants was recorded due to incorporation of straw alongwith the micro-organisms in soil. The organic carbon and nitrogen content of the soil increased with all the treatments except control. The highest increase in organic carbon and nitrogen of the soil was observed with a treatment of wheat straw of 65 C:P ratio inoculated withS. coccosporum. The yield of wheat crop after groundnut was significantly more with several treatments than control plots. The highest increase of 79 per cent in grain yield of wheat was observed in the plots previouslq received with wheat straw of 200 C:P ratio.This paper is based on the data presented at IV Southern Regional Conference on Microbial Inoculants, held at Parbhani during 3–4 July 1978.     

餐厨垃圾厌氧消化产沼气过程中酶学表征

厌氧消化产沼气被认为是餐厨垃圾资源化利用的有效方式之一,其实质是在多种微生物综合作用下的生物化学过程.本文研究了在促进和抑制性因子作用下,餐厨垃圾厌氧发酵的酶学过程,对其中的脱氢酶和水解酶(β-葡萄糖苷酶,BAA-蛋白水解酶,碱性磷酸酶)活性变化进行了分析.研究表明,与空白对照组相比添加酵母粉后脱氢酶的最高活性提高了8...     

The Survival of antibiotic resistant and sensitiveEscherichia coli strains during anaerobic digestion

Summary Total counts ofEscherichia coli were followed during anaerobic digestion of pig slurry laboratory scale digesters at 37° C. Counts decreased rapidly during anaerobic digestion. Antibiotic resistant strains in most cases appeared to be more persistent in anaerobic digesters than sensitiveE. coli strains as calculated from the decimal decay rates.     

Fungal pretreatment: An alternative in second-generation ethanol from wheat straw

The potential of a fungal pretreatment combined with a mild alkali treatment to replace or complement current physico-chemical methods for ethanol production from wheat straw has been investigated. Changes in substrate composition, secretion of ligninolytic enzymes, enzymatic hydrolysis efficiency and ethanol yield after 7, 14 and 21 days of solid-state fermentation were evaluated. Most fungi degraded lignin with variable selectivity degrees, although only eight of them improved sugar recovery compared to untreated samples. Glucose yield after 21 days of pretreatment with Poria subvermispora and Irpex lacteus reached 69% and 66% of cellulose available in the wheat straw, respectively, with an ethanol yield of 62% in both cases. Conversions from glucose to ethanol reached around 90%, showing that no inhibitors were generated during this pretreatment. No close correlations were found between ligninolytic enzymes production and sugar yields.     

Enhanced ethanol and chitosan production from wheat straw by Mucor indicus with minimal nutrient consumption

Recently, Mucor indicus was introduced as a promising ethanol producing microorganism for fermentation of lignocellulosic hydrolysates, showing a number of advantages over Saccharomyces cerevisiae. However, high nutrient requirement is the main drawback of the fungus in efficient ethanol production from lignocelluloses. In this study, application of fungal extract as a potential nutrient source replacing all required nutrients in fermentation of wheat straw by M. indicus was investigated. Wheat straw was pretreated with N-methylmorpholine-N-oxide (NMMO) at 120 °C for 1–5 h prior to enzymatic hydrolysis. Hydrolysis yield was improved at least by 6-fold for 3 h pretreated straw compared with that of untreated one. A fungal extract was produced by autolysis of M. indicus biomass, an unavoidable byproduct of fermentation. Maximum free amino nitrogen (2.04 g/L), phosphorus (1.50 g/L), and total nitrogen (4.47 g/L) as well as potassium, magnesium, and calcium in the fungal extract were obtained by autolysis of the biomass at 50 °C and pH 5.0. The fungal extract as a nutrient-rich supplement substituted yeast extract and all other required minerals in fermentation and enhanced the ethanol yield up to 92.1% of the theoretical yield. Besides, appreciate amounts of chitosan were produced as another valuable product of the autolysis.     

Anaerobic digestion of secondary residuals from an anaerobic bioreactor at a brewery to enhance bioenergy generation

Many beer breweries use high-rate anaerobic digestion (AD) systems to treat their soluble high-strength wastewater. Biogas from these AD systems is used to offset nonrenewable energy utilization in the brewery. With increasing nonrenewable energy costs, interest has mounted to also digest secondary residuals from the high-rate digester effluent, which consists of yeast cells, bacteria, methanogens, and small (hemi)cellulosic particles. Mesophilic (37 °C) and thermophilic (55 °C) lab-scale, low-rate continuously-stirred anaerobic digestion (CSAD) bioreactors were operated for 258 days by feeding secondary residuals at a volatile solids (VS) concentration of ∼40 g l⁻¹. At a hydraulic retention time (HRT) of 15 days and a VS loading rate of 2.7 g VS l⁻¹ day⁻¹, the mesophilic bioreactor showed an average specific volumetric biogas production rate of 0.88 l CH₄ l⁻¹ day⁻¹ and an effluent VS concentration of 22.2 g VS l⁻¹ (43.0% VS removal efficiency) while the thermophilic bioreactor displayed similar performances. The overall methane yield for both systems was 0.21 l CH₄ g⁻¹ VS fed and 0.47–0.48 l CH₄ g⁻¹ VS removed. A primary limitation of thermophilic digestion of this protein-rich waste is the inhibition of methanogens due to higher nondissociated (free) ammonia (NH₃) concentrations under similar total ammonium (NH₄ ⁺) concentrations at equilibrium. Since thermophilic AD did not result in advantageous methane production rates or yields, mesophilic AD was, therefore, superior in treating secondary residuals from high-rate AD effluent. An additional digester to convert secondary residuals to methane may increase the total biogas generation at the brewery by 8% compared to just conventional high-rate digestion of brewery wastewater alone. JIMB-2008: BioEnergy—Special issue.     

Enhancing the solid-state anaerobic digestion of fallen leaves through simultaneous alkaline treatment

Previous studies have shown that alkali pretreatment prior to anaerobic digestion (AD) can increase the digestibility of lignocellulosic biomass and methane yield. In order to simplify the process and reduce the capital cost, simultaneous alkali treatment and anaerobic digestion was evaluated for methane production from fallen leaves. The highest methane yield of 82 L/kg volatile solids (VS) was obtained at NaOH loading of 3.5% and substrate-to-inoculum (S/I) ratio of 4.1. The greatest enhancement in methane yield was achieved at S/I ratio of 6.2 with NaOH loading of 3.5% which was 24-fold higher than that of the control (without NaOH addition). Reactors at S/I ratio of 8.2 resulted in failure of the AD process. In addition, increasing the total solid (TS) content from 20% to 26% reduced biogas yield by 35% at S/I ratio of 6.2 and NaOH loading of 3.5%. Cellulose and hemicellulose degradation and methane yields are highly related.     

Kinetic model of enzymatic hydrolysis of steam-exploded wheat straw

The hydrolysis kinetics of steam-exploded wheat straw treated with cellulase NS 50013 enzyme complex in combination with β-glucosidase NS 50010 is studied. The time dependence of the reducing sugars amount is followed at varying the temperature value and the amount of the enzyme introduced. The activation energy determined on the ground of the rate temperature dependence stays unchanged in the course of the process. The preexponential factor decreases with the increase of the degree of hydrolysis and is responsible for the process rate decrease. A new expression for the dependence of degree of hydrolysis of one of carbohydrate polymers (cellulose) in wheat straw on the time, the enzyme concentration and the temperature is obtained. It is of practical importance as well because it provides estimation of the degree of hydrolysis required at predetermined values of the temperature, the enzyme concentration and the time used. The expression can be used for control of the enzyme hydrolysis of cellulose in the wheat straw.     

碳氮比对模拟水稻秸秆田间厌氧发酵系统甲烷生产和秸秆降解的影响

利用自制的厌氧发酵装置模拟田间厌氧发酵产生物甲烷的条件,通过添加不同量的尿素,研究C/N对水稻秸秆厌氧发酵产生物甲烷和秸秆降解的影响。设置了C/N为15∶1、20∶1、25∶1、30∶1和对照(53∶1)等五个处理,测定各处理的产气量、甲烷含量和秸秆木质纤维素含量。结果表明C/N对厌氧发酵产气进程有一定影响,峰值出现的时间、峰值的大小在不同处理之间均有差异:以C/N为25∶1的处理峰值出现的最早,较对照早5 d出现;以C/N为20∶1的峰值最高,较对照提高了27. 55%。C/N对厌氧发酵累积产气量和累积甲烷产量有显著影响,以C/N为20∶1的处理累积产气量(4 235. 00 mL)和累积甲烷产量(1526. 34mL)最高,相比对照组的累积产气量和累积甲烷产量分别提高了30. 78%和51. 31%。C/N对水稻秸秆降解也有显著影响。以20∶1和25∶1处理对秸秆降解的效果最好,其中20∶1处理的总降解率、纤维素降解率和半纤维素降解率分别达到了51. 33%、55. 31%和53. 05%,25∶1处理分别为50. 00%、53. 08%和49. 42%。上述结果表明通过添加尿素调节C/N能够提高水稻秸秆厌氧发酵的产生物甲烷效率和促进秸秆降解,以C/N为(20～25)∶1处理的促进效果最好。     

Modification of wheat straw lignin by solid state fermentation with white-rot fungi

The potential of crude enzyme extracts, obtained from solid state cultivation of four white-rot fungi (Trametes versicolor, Bjerkandera adusta, Ganoderma applanatum and Phlebia rufa), was exploited to modify wheat straw cell wall. At different fermentation times, manganese-dependent peroxidase (MnP), lignin peroxidase (LiP), laccase, carboxymethylcellulase (CMCase), avicelase, xylanase and feruloyl esterase activities were screened and the content of lignin as well as hydroxycinnamic acids in fermented straw were determined. All fungi secreted feruloyl esterase while LiP was only detected in crude extracts from B. adusta. Since no significant differences (P > 0.05) were observed in remaining lignin content of fermented straw, LiP activity was not a limiting factor of enzymatic lignin removal process. The levels of esterified hydroxycinnamic acids degradation were considerably higher than previous reports with lignocellulosic biomass. The data show that P. rufa, may be considered for more specific studies as higher ferulic and p-coumaric acids degradation was observed for earlier incubation times.