The exploitation of municipal solid waste (MSW) and related waste paper streams in the production of bioalcohol

An organic fraction from municipal solid waste (MSW) comprised 38.9% (w/w) glucose (cellulose and starch) indicating its potential as a substrate for bioalcohol production. Microscopy indicated that the fraction was rich in waste paper fibres. Much paper waste comes from shredded office paper (50.4% w/w glucose) which is unrecyclable because of poor fibre length. This, and microbiological hazards associated with the use of MSW led to its choice as model substrate for study. Saccharification of shredded paper waste was optimised by selection of Accellerase® and additional beta-glucosidase enabling digestion of 99.27% of cellulose. Sequential batch-addition of substrate permitted substrate “concentrations” equivalent to 25-30% (w/v). Saccharification was enhanced by detergent, but reduced by the presence of alcohols at over 3-4% (v/v). Steam explosion of paper slightly enhanced saccharification. However, the approach was rejected due to high energy cost, production of fermentation inhibitors at high severities, and lack of clear benefit regarding ethanol yield. Interestingly, levels of inhibitors were low compared to other pre-treated substrates and addition of paper to other substrates greatly reduced their own production of inhibitors during pre-treatment (wheat straw 60%, filter paper 95%). Larger pilot-scale (1.5-5 L) operations involved developing the batch-addition regime with a high-shear stirring capacity vessel. Additions equating to final substrate concentrations of ~65% (w/v) were achieved (from an initial 5% w/v) and facilitated high ethanol concentrations (11.6% v/v) with minimal enzyme input (3.7 FPU/g substrate). Thermal tolerance of a range of yeast strains was investigated by developing a rapid screening approach with liquid-handling robotics. This identified strains able to endure temperatures up to 40°C. Evolutionary engineering may improve tolerances to temperatures nearer to enzyme optimums (50°C). Some previously unused strains exhibited superior growth to referenced industrial strains. The above findings were integrated into a process design along with recommendations for further enhancement

Release of cell wall phenolic esters during hydrothermal pretreatment of rice husk and rice straw

Background: Rice husk and rice straw represent promising sources of biomass for production of renewable fuels and chemicals. For efficient utilisation, lignocellulosic components must first be pretreated to enable efficient enzymatic saccharification and subsequent fermentation. Existing pretreatments create breakdown products such as sugar-derived furans, and lignin-derived phenolics that inhibit enzymes and fermenting organisms. Alkali pretreatments have also been shown to release significant levels of simple, free phenolics such as ferulic acid that are normally esterified to cell wall polysaccharides in the intact plant. These phenolics have recently been found to have considerable inhibitory properties. The aim of this research has been to establish the extent to which such free phenolic acids are also released during hydrothermal pretreatment of rice straw (RS) and rice husk (RH). Results: RS and RH were subjected to hydrothermal pretreatments over a wide range of severities (1.57–5.45). FTIR analysis showed that the pretreatments hydrolysed and solubilised hemicellulosic moieties, leading to an enrichment of lignin and crystalline cellulose in the insoluble residue. The residues also lost the capacity for UV autofluorescence at pH 7 or pH 10, indicating the breakdown or release of cell wall phenolics. Saponification of raw RS and RH enabled identification and quantification of substantial levels of simple phenolics including ferulic acid (tFA), coumaric acid (pCA) and several diferulic acids (DiFAs) including 8-O-4′-DiFA, 8,5′-DiFA and 5,5′-DiFA. RH had higher levels of pCA and lower levels of tFA and DiFAs compared with RS. Assessment of the pretreatment liquors revealed that pretreatment-liberated phenolics present were not free but remained as phenolic esters (at mM concentrations) that could be readily freed by saponification. Many were lost, presumably through degradation, at the higher severities. Conclusion: Differences in lignin, tFA, DiFAs and pCA between RS and RH reflect differences in cell wall physiology, and probably contribute to the higher recalcitrance of RH compared with RS. Hydrothermal pretreatments, unlike alkali pretreatments, release cinnamic acid components as esters. The potential for pretreatment-liberated phenolic esters to be inhibitory to fermenting microorganisms is not known. However, the present study shows that they are found at concentrations that could be significantly inhibitory if released as free forms by enzyme activity

Characterisation of lignocellulosic sugars from municipal solid waste residue.

Municipal solid waste (MSW) contains significant quantities of plant-derived carbohydrates which have the potential to be exploited as a biomass source. This study evaluated the chemical composition and fractionation of MSW water-insoluble organic matter remaining after recycling of other components (MSWR). The organic matter was prepared as a dry, alcohol insoluble residue (MSWR-AIR, comprising w = 6% of original MSW) and size fractionated into fractions A, B, C & D. Carbohydrates were present in all the sub-fractions, comprising up to w = 54%; their complexity was also assessed by FT-IR spectroscopy. The lignin content in the samples ranged from w = 11–22%. The most carbohydrate-rich subfraction (C; w = 4% original MSW) was sequentially extracted to provide information on the likely constituent cell wall-derived polymers, sugar compositions and uronic acid content. The results indicate that approximately w = 25% of the MSWR-AIR comprises glucose, which appears to be mostly cellulosic in origin. The results are discussed in relation to the potential for exploitation

Optimising conditions for bioethanol production from rice husk and rice straw: effects of pre-treatment on liquor composition and fermentation inhibitors.

BACKGROUND: Rice straw and husk are globally significant sources of cellulose-rich biomass and there is great interest in converting them to bioethanol. However, rice husk is reportedly much more recalcitrant than rice straw and produces larger quantities of fermentation inhibitors. The aim of this study was to explore the underlying differences between rice straw and rice husk with reference to the composition of the pre-treatment liquors and their impacts on saccharification and fermentation. This has been carried out by developing quantitative NMR screening methods. RESULTS: Air-dried rice husk and rice straw from the same cultivar were used as substrates. Carbohydrate compositions were similar, whereas lignin contents differed significantly (husk: 35.3% w/w of raw material; straw 22.1% w/w of raw material). Substrates were hydrothermally pre-treated with high-pressure microwave processing across a wide range of severities. 25 compounds were identified from the liquors of both pre-treated rice husk and rice straw. However, the quantities of compounds differed between the two substrates. Fermentation inhibitors such as 5-HMF and 2-FA were highest in husk liquors, and formic acid was higher in straw liquors. At a pre-treatment severity of 3.65, twice as much ethanol was produced from rice straw (14.22% dry weight of substrate) compared with the yield from rice husk (7.55% dry weight of substrate). Above severities of 5, fermentation was inhibited in both straw and husk. In addition to inhibitors, high levels of cellulase-inhibiting xylo-oligomers and xylose were found and at much higher concentrations in rice husk liquor. At low severities, organic acids and related intracellular metabolites were released into the liquor. CONCLUSIONS: Rice husk recalcitrance to saccharification is probably due to the much higher levels of lignin and, from other studies, likely high levels of silica. Therefore, if highly polluting chemical pre-treatments and multi-step biorefining processes are to be avoided, rice husk may need to be improved through selective breeding strategies, although more careful control of pre-treatment may be sufficient to reduce the levels of fermentation inhibitors, e.g. through steam explosion-induced volatilisation. For rice straw, pre-treating at severities of between 3.65 and 4.25 would give a glucose yield of between 37.5 and 40% (w/DW, dry weight of the substrate) close to the theoretical yield of 44.1% w/DW, and an insignificant yield of total inhibitors

Biorefining of waste paper biomass: increasing the concentration of glucose by optimising enzymatic hydrolysis

Waste copier paper is a potential substrate for the production of glucose relevant for manufacture of platform chemicals and intermediates, being composed of 51 % glucan. The yield and concentration of glucose arising from the enzymatic saccharification of solid ink-free copier paper as cellulosic substrate was studied using a range of commercial cellulase preparations. The results show that in all cellulase preparations examined, maximum hydrolysis was only achieved with the addition of beta-glucosidase, despite its presence in the enzyme mixtures. With the use of AccelleraseA (R) (cellulase), high substrate loading decreased conversion yield. However, this was overcome if the enzyme was added between 12.5 and 20 FPU g substrate(-1). Furthermore, this reaction condition facilitated continual stirring and enabled sequential additions (up to 50 % w/v) of paper to be made to the hydrolysis reaction, degrading nearly all (99 %) of the cellulose fibres and increasing the final concentration of glucose whilst simultaneously making high substrate concentrations achievable. Under optimal conditions (50 A degrees C, pH 5.0, 72 h), digestions facilitate the production of glucose to much improved concentrations of up to 1.33 mol l(-1)

Bioethanol production from spent mushroom compost derived from chaff of millet and sorghum

Abstract Background In Uganda, the chaff remaining from threshed panicles of millet and sorghum is a low value, lignocellulose-rich agricultural by-product. Currently, it is used as a substrate for the cultivation of edible Oyster mushrooms (Pleurotus ostreatus). The aim of this study was to assess the potential to exploit the residual post-harvest compost for saccharification and fermentation to produce ethanol. Results Sorghum and millet chaff-derived spent oyster mushroom composts minus large mycelium particles were assessed at small-scale and low substrate concentrations (5% w/v) for optimal severity hydrothermal pre-treatment, enzyme loading and fermentation with robust yeasts to produce ethanol. These conditions were then used as a basis for larger scale assessments with high substrate concentrations (30% w/v). Millet-based compost had a low cellulose content and, at a high substrate concentration, did not liquefy effectively. The ethanol yield was 63.9 g/kg dry matter (DM) of original material with a low concentration (19.6 g/L). Compost derived from sorghum chaff had a higher cellulose content and could be liquefied at high substrate concentration (30% w/v). This enabled selected furfural-resistant yeasts to produce ethanol at up to 186.9 g/kg DM of original material and a concentration of 45.8 g/L. Conclusions Spent mushroom compost derived from sorghum chaff has the potential to be an industrially useful substrate for producing second-generation bioethanol. This might be improved further through fractionation and exploitation of hemicellulosic moieties, and possibly the exploitation of the mycelium-containing final residue for animal feed. However, spent compost derived from millet does not provide a suitably high concentration of ethanol to make it industrially attractive. Further research on the difficulty in quantitatively saccharifying cellulose from composted millet chaff and other similar substrates such as rice husk is required

Characterization of cell wall components of wheat straw following hydrothermal pretreatment and fractionation

Thermophysical pretreatment enhances the enzymatic hydrolysis of lignocellulose. However, its impact on cell wall chemistry is still poorly understood. This paper reports the effects of hydrothermal pretreatment on the degradation and alkali-extractability of wheat straw cell wall polymers. Pretreatment resulted in loss and/or solubilization of arabinoxylans (by 53%), ferulic and diferulic acids which are important cross-linking agents accompanied by concomitant increases in cellulose (up to 43%) and lignin (29%). The remaining water-insoluble hemicelluloses were more readily extractable in alkali and were reduced in molecular weight indicating substantial thermochemical depolymerization. They were also associated with smaller but significant amounts of (cellulose-derived) glucose. The alkali-insoluble residues consisted predominantly of cellulosic glucose and lignin and contained p-coumaric acid. The depolymerization of hemicelluloses, reduction in cinnamic acids and partial degradation of cellulose is likely to contribute significantly to the accessibility of cellulases during subsequent enzymolysis. (C) 2012 Elsevier Ltd. All rights reserved