Simultaneous saccharification and bioethanol production from corn cobs: Process optimization and kinetic studies.

Science.gov (United States)

Sewsynker-Sukai, Yeshona; Gueguim Kana, E B

2018-08-01

This study investigates the simultaneous saccharification and fermentation (SSF) process for bioethanol production from corn cobs with prehydrolysis (PSSF) and without prehydrolysis (OSSF). Two response surface models were developed with high coefficients of determination (>0.90). Process optimization gave high bioethanol concentrations and bioethanol conversions for the PSSF (36.92 ± 1.34 g/L and 62.36 ± 2.27%) and OSSF (35.04 ± 0.170 g/L and 58.13 ± 0.283%) models respectively. Additionally, the logistic and modified Gompertz models were used to study the kinetics of microbial cell growth and ethanol formation under microaerophilic and anaerobic conditions. Cell growth in the OSSF microaerophilic process gave the highest maximum specific growth rate (µ max ) of 0.274 h -1 . The PSSF microaerophilic bioprocess gave the highest potential maximum bioethanol concentration (P m ) (42.24 g/L). This study demonstrated that microaerophilic rather than anaerobic culture conditions enhanced cell growth and bioethanol production, and that additional prehydrolysis steps do not significantly impact on the bioethanol concentration and conversion in SSF process. Copyright © 2018 Elsevier Ltd. All rights reserved.

Kinetic modeling of simultaneous saccharification and fermentation of corn starch for ethanol production.

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Białas, Wojciech; Czerniak, Adrian; Szymanowska-Powałowska, Daria

2014-01-01

Fuel ethanol production, using a simultaneous saccharification and fermentation process (SSF) of native starch from corn flour, has been performed using Saccharomyces cerevisiae and a granular starch hydrolyzing enzyme. The quantitative effects of mash concentration, enzyme dose and pH were investigated with the use of a Box-Wilson central composite design protocol. Proceeding from results obtained in optimal fermentation conditions, a kinetics model relating the utilization rates of starch and glucose as well as the production rates of ethanol and biomass was tested. Moreover, scanning electron microscopy (SEM) was applied to investigate corn starch granule surface after the SFF process. A maximum ethanol concentration of 110.36 g/l was obtained for native corn starch using a mash concentration of 25%, which resulted in ethanol yield of 85.71%. The optimal conditions for the above yield were found with an enzyme dose of 2.05 ml/kg and pH of 5.0. These results indicate that by using a central composite design, it is possible to determine optimal values of the fermentation parameters for maximum ethanol production. The investigated kinetics model can be used to describe SSF process conducted with granular starch hydrolyzing enzymes. The SEM micrographs reveal randomly distributed holes on the surface of granules.

Production of ethanol from a mixture of waste paper and kitchen waste via a process of successive liquefaction, presaccharification, and simultaneous saccharification and fermentation.

Science.gov (United States)

Nishimura, Hiroto; Tan, Li; Kira, Noriko; Tomiyama, Shigeo; Yamada, Kazuo; Sun, Zhao-Yong; Tang, Yue-Qin; Morimura, Shigeru; Kida, Kenji

2017-09-01

Efficient ethanol production from waste paper requires the addition of expensive nutrients. To reduce the production cost of ethanol from waste paper, a study on how to produce ethanol efficiently by adding kitchen waste (potentially as a carbon source, nutrient source, and acidity regulator) to waste paper was performed and a process of successive liquefaction, presaccharification, and simultaneous saccharification and fermentation (L+PSSF) was developed. The individual saccharification performances of waste paper and kitchen waste were not influenced by their mixture. Liquefaction of kitchen waste at 90°C prior to presaccharification and simultaneous saccharification and fermentation (PSSF) was essential for efficient ethanol fermentation. Ethanol at concentrations of 46.6 or 43.6g/l was obtained at the laboratory scale after fermentation for 96h, even without pH adjustment and/or the addition of extra nutrients. Similarly, ethanol at a concentration of 45.5g/l was obtained at the pilot scale after fermentation for 48h. The ethanol concentration of L+PSSF of the mixture of waste paper and kitchen waste was comparable to that of PSSF of waste paper with added nutrients (yeast extract and peptone) and pH adjustment using H 2 SO 4 , indicating that kitchen waste is not only a carbon source but also an excellent nutrient source and acidity regulator for fermentation of the mixture of waste paper and kitchen waste. Copyright © 2017. Published by Elsevier Ltd.

Lactic acid production from unmatured banana peel and flesh through simultaneous saccharification and fermentation

Directory of Open Access Journals (Sweden)

Mohammed BELMAKKI

2016-07-01

Full Text Available The aim of this study was to establish a process of lactic acid (LA production from two different kinds of african organic waste i.e. peel and flesh of un-matured banana by using as model strain Lactobacillus bp Pentosus AH 239. The bioconversion of glucose contained in the biomass to LA was performed following the Simultaneous Saccharification and Fermentation (SSF process. The Separated Hydrolysis and Fermentation (SHF was also applied in this study to compare the efficiency of both process. The results showed that the enzymatic hydrolysis yield was significantly improved in case of SSF recording a rate of hydrolysis in the range of 82%-90% against 52%-61% under SHF conditions. The results showed also that SSF give more efficient lactic acid production with a yield above of 90%, and a high concentration up to 50 g/L. Due to its performance, the SSF process for the lactic acid production could be an important way of bioconversion for lignocellulosic residues in Africa. The optimization of this process need to be adapted for African context and for its development on an industrial scale.

Kinetic modeling of multi-feed simultaneous saccharification and co-fermentation of pretreated birch to ethanol.

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Wang, Ruifei; Koppram, Rakesh; Olsson, Lisbeth; Franzén, Carl Johan

2014-11-01

Fed-batch simultaneous saccharification and fermentation (SSF) is a feasible option for bioethanol production from lignocellulosic raw materials at high substrate concentrations. In this work, a segregated kinetic model was developed for simulation of fed-batch simultaneous saccharification and co-fermentation (SSCF) of steam-pretreated birch, using substrate, enzymes and cell feeds. The model takes into account the dynamics of the cellulase-cellulose system and the cell population during SSCF, and the effects of pre-cultivation of yeast cells on fermentation performance. The model was cross-validated against experiments using different feed schemes. It could predict fermentation performance and explain observed differences between measured total yeast cells and dividing cells very well. The reproducibility of the experiments and the cell viability were significantly better in fed-batch than in batch SSCF at 15% and 20% total WIS contents. The model can be used for simulation of fed-batch SSCF and optimization of feed profiles. Copyright © 2014 Elsevier Ltd. All rights reserved.

Comparison of bio-hydrogen production yield capacity between asynchronous and simultaneous saccharification and fermentation processes from agricultural residue by mixed anaerobic cultures.

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Li, Yameng; Zhang, Zhiping; Zhu, Shengnan; Zhang, Huan; Zhang, Yang; Zhang, Tian; Zhang, Quanguo

2018-01-01

Taken common agricultural residues as substrate, dark fermentation bio-hydrogen yield capacity from asynchronous saccharification and fermentation (ASF) and simultaneous saccharification and fermentation (SSF) was investigated. The highest hydrogen yield of 472.75mL was achieved with corncob using ASF. Hydrogen yield from corn straw, rice straw, corncob and sorghum stalk by SSF were 20.54%,10.31%,13.99% and 5.92% higher than ASF, respectively. The experimental data fitted well to the modified Gompertz model. SSF offered a distinct advantage over ASF with respect to reducing overall process time (60h of SSF, 108h of ASF). Meanwhile, SSF performed better than SSF with respect to shortening the lag-stage. The major metabolites of anaerobic fermentation hydrogen production by ASF and SSF were butyric acid and acetic acid. Copyright © 2017 Elsevier Ltd. All rights reserved.

A MICROWAVE-ASSISTED LIQUEFACTION AS A PRETREATMENT FOR THE BIOETHANOL PRODUCTION BY THE SIMULTANEOUS SACCHARIFICATION AND FERMENTATION OF CORN MEAL

Directory of Open Access Journals (Sweden)

Svetlana Nikolić

2008-11-01

Full Text Available A microwave-assisted liquefaction as a pretreatment for the bioethanol production by the simultaneous saccharification and fer entation (SSF of corn meal using Saccharomyces cerevisiae var. ellipsoideus yeast in a batch system was studied. An optimal power of microwaves of 80 W and the 5-min duration of the microwave treatment were selected by following the concentration of glucose released from the corn meal suspensions at hidromodul of 1:3 (corn meal to water ratio in the liquefaction step. The results indicated that the microwave pretreatment could increase the maximum ethanol concentration produced in the SSF process for 13.4 %. Consequently, a significant increase of the ethanol productivity on substrate (YP/S, as well as the volumetric ethanol productivity (P in this process, could be achieved

Production of L- and D-lactic acid from waste Curcuma longa biomass through simultaneous saccharification and cofermentation.

Science.gov (United States)

Nguyen, Cuong Mai; Kim, Jin-Seog; Nguyen, Thanh Ngoc; Kim, Seul Ki; Choi, Gyung Ja; Choi, Yong Ho; Jang, Kyoung Soo; Kim, Jin-Cheol

2013-10-01

Simultaneous saccharification and cofermentation (SSCF) of Curcuma longa waste biomass obtained after turmeric extraction to L- and D-lactic acid by Lactobacillus coryniformis and Lactobacillus paracasei, respectively, was investigated. This is a rich, starchy, agro-industrial waste with potential for use in industrial applications. After optimizing the fermentation of the biomass by adjusting nitrogen sources, enzyme compositions, nitrogen concentrations, and raw material concentrations, the SSCF process was conducted in a 7-l jar fermentor at 140 g dried material/L. The maximum lactic acid concentration, average productivity, reducing sugar conversion and lactic acid yield were 97.13 g/L, 2.7 g/L/h, 95.99% and 69.38 g/100 g dried material for L-lactic acid production, respectively and 91.61 g/L, 2.08 g/L/h, 90.53% and 65.43 g/100 g dried material for D-lactic acid production, respectively. The simple and efficient process described in this study could be utilized by C. longa residue-based lactic acid industries without requiring the alteration of plant equipment. Copyright © 2013 Elsevier Ltd. All rights reserved.

Development of a semi-continuous two-stage simultaneous saccharification and fermentation process for enhanced 2,3-butanediol production by Klebsiella oxytoca.

Science.gov (United States)

Moon, S-K; Kim, D-K; Park, J M; Min, J; Song, H

2018-04-01

Klebsiella oxytoca naturally produces a large amount of 2,3-butanediol (2,3-BD), a promising chemical with wide industrial applications, along with various by-products. Previously, we have developed a metabolically engineered K. oxytoca ΔldhA ΔpflB strain to reduce the formation of by-products. To improve 2,3-BD productivity and examine the stability of K. oxytoca ΔldhA ΔpflB strain for industrial application, a semi-continuous two-stage simultaneous saccharification and fermentation (STSSF) process was developed. The STSSF with the K. oxytoca ΔldhA ΔpflB mutant using cassava as a carbon source could produce 108 ± 3·73 g (2,3- BD )  l -1 with a yield of 0·45 g (2,3- BD )  g (glucose) -1 and a productivity of 3·00 g (2,3- BD ) l -1  h -1 . No apparent changes in the final titre, yield and productivity of 2,3-BD were observed for up to 20 cycles of STSSF. Also, microbial contamination and spontaneous mutation of the host strain with potential detrimental effects on fermentation efficiency did not occur during the whole fermentation period. These results strongly underpin that the K. oxytoca ΔldhA ΔpflB mutant is stable and that the STSSF process is commercially exploitable. There is growing interest in the production of 2,3-butanediol (2,3-BD) from renewable resources by microbial fermentation because of its wide applications to specialty and commodity chemical industries. Klebsiella oxytoca usually produces 2,3-BD as a major end product during the fermentation of carbohydrates. This is the first study to provide a high-efficiency simultaneous saccharification and 2,3-BD fermentation process. Also, this study proves the stability of a metabolically engineered 2,3-BD overproducing K. oxytoca strain for industrial application. © 2018 The Society for Applied Microbiology.

Biological Hydrogen Production: Simultaneous Saccharification and Fermentation with Nitrogen and Phosphorus Removal from Wastewater Effluent

Science.gov (United States)

2012-03-01

process.7 The reaction is of great economic importance given that the world’s industrial production of nitrogenous fertilizer increased 27-fold between... Enzymatic Saccharification and Fermentation of Paper and Pulp Industry Effluent for Biohydrogen Production . Int. J. Hydrogen Energy 2010, 35, pp...Reactor Setup and Operation 11 4.2 Operational Comparison: SBR and CBR 12 4.3 Effect of pH and Loading on Hydrogen Production 13 4.4 Enzymatic Source

Butanol production from wheat straw by simultaneous saccharification and fermentation using Clostridium beijerinckii: Part II-Fed-batch fermentation

International Nuclear Information System (INIS)

Qureshi, Nasib; Saha, Badal C.; Cotta, Michael A.

2008-01-01

In these studies, Clostridium beijerinckii P260 was used to produce butanol (acetone-butanol-ethanol, or ABE) from wheat straw (WS) hydrolysate in a fed-batch reactor. It has been demonstrated that simultaneous hydrolysis of WS to achieve 100% hydrolysis to simple sugars (to the extent achievable under present conditions) and fermentation to butanol is possible. In addition to WS, the reactor was fed with a sugar solution containing glucose, xylose, arabinose, galactose, and mannose. The culture utilized all of the above sugars. It was noticed that near the end of fermentation (286-533 h), the culture had difficulties utilizing xylose. As a result of supplemental sugar feed to the reactor, ABE productivity was improved by 16% as compared with previous studies. In our previous experiment on simultaneous saccharification of WS and fermentation to butanol, a productivity of 0.31 g L -1 h -1 was observed, while in the present studies a productivity of 0.36 g L -1 h -1 was observed. It should be noted that a productivity of 0.77 g L -1 h -1 was observed when the culture was highly active. The fed-batch fermentation was operated for 533 h. It should be noted that C. beijerinckii P260 can be used to produce butanol from WS in integrated fermentations

High titer ethanol production from SPORL-pretreated lodgepole pine by simultaneous enzymatic saccharification and combined fermentation.

Science.gov (United States)

Lan, T Q; Gleisner, Roland; Zhu, J Y; Dien, Bruce S; Hector, Ronald E

2013-01-01

Lodgepole wood chips were pretreated by sulfite pretreatment to overcome recalcitrance of lignocelluloses (SPORL) at 25% solids loading and 180 °C for 20 min with sulfuric acid and sodium bisulfite charges of 2.2 and 8 wt/wt% on an oven-dry wood basis, respectively. The pretreated wood chips were disk-milled with pretreatment spent liquor and water, and the solid fraction was separated from the liquor stream. The liquor was neutralized and concentrated through vacuum evaporation. Quasi-simultaneous enzymatic saccharification of the cellulosic solids and combined fermentation with the concentrated liquor was conducted at up to 20% total solids loading. Fed-batching of the solids facilitated liquefaction and saccharification, as well as managing instantaneous inhibitor concentrations. At a commercial cellulase (CTec2) loading of only 9 FPU or 0.06 mL/g untreated wood, a maximum ethanol titer of 47.4 g/L was achieved, resulting in a calculated yield of 285 L/tonne of wood using Saccharomyces cerevisiae YRH400 at 35 °C and pH 5.5. Published by Elsevier Ltd.

Optimization of enzymatic hydrolysis for ethanol production by simultaneous saccharification and fermentation of wastepaper.

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Sangkharak, Kanokphorn

2011-11-01

The present study investigated the development of high sugar production by optimization of an enzymatic hydrolysis process using both conventional and statistical methods, as well as the production of ethanol by the selected wastepaper source. Among four sources of pretreated wastepaper including office paper, newspaper, handbills and cardboard, office paper gave the highest values of cellulose (87.12%) and holocelluloses (89.07%). The effects of the amount of wastepaper, the pretreatment method and the type of enzyme on reducing sugar production from office paper were studied using conventional methods. The highest reducing sugar production (1851.28 µg L(-1); 37.03% conversion of glucose) was obtained from the optimal condition containing 40 mg of office paper, pretreated with stream explosion and hydrolysed with the combination of cellulase from Aspergillus niger and Trichoderma viride at the fixed loading rate of 20 FPU g(-1) sample. The effects of interaction of wastepaper amount and enzyme concentration as well as incubation time were studied by a statistical method using central composite design. The optimal medium composition consisted of 43.97 µg L(-1), 28.14 FPU g(-1) sample and 53.73 h of wastepaper, enzyme concentration and incubation time, respectively, and gave the highest amount of sugar production (2184.22 µg L(-1)) and percentage conversion of glucose (43.68%). The ethanol production from pretreated office paper using Saccharomyces cerevisiae in a simultaneous saccharification and fermentation process was 21.02 g L(-1) after 36 h of cultivation, corresponding to an ethanol volumetric production rate of 0.58 g ethanol L(-1) h(-1).

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

Energy Technology Data Exchange (ETDEWEB)

Srichuwong, Sathaporn; Fujiwara, Maki; Wang, Xiaohui; Seyama, Tomoko; Shiroma, Riki; Arakane, Mitsuhiro; Tokuyasu, Ken [National Food Research Institute, National Agriculture and Food Research Organization (NARO), 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642 (Japan); Mukojima, Nobuhiro [National Agricultural Research Center for Hokkaido Region, NARO, 9-4 Shinsei-minami, Memuro-cho, Kasai-gun, Hokkaido 082-0071 (Japan)

2009-05-15

Simultaneous saccharification and fermentation (SSF) of very high gravity (VHG) potato mash, containing 304 g L{sup -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 {alpha}-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{sup -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. (author)

Modeling and parameter identification of the simultaneous saccharification-fermentation process for ethanol production.

Science.gov (United States)

Ochoa, Silvia; Yoo, Ahrim; Repke, Jens-Uwe; Wozny, Günter; Yang, Dae Ryook

2007-01-01

Despite many environmental advantages of using alcohol as a fuel, there are still serious questions about its economical feasibility when compared with oil-based fuels. The bioethanol industry needs to be more competitive, and therefore, all stages of its production process must be simple, inexpensive, efficient, and "easy" to control. In recent years, there have been significant improvements in process design, such as in the purification technologies for ethanol dehydration (molecular sieves, pressure swing adsorption, pervaporation, etc.) and in genetic modifications of microbial strains. However, a lot of research effort is still required in optimization and control, where the first step is the development of suitable models of the process, which can be used as a simulated plant, as a soft sensor or as part of the control algorithm. Thus, toward developing good, reliable, and simple but highly predictive models that can be used in the future for optimization and process control applications, in this paper an unstructured and a cybernetic model are proposed and compared for the simultaneous saccharification-fermentation process (SSF) for the production of ethanol from starch by a recombinant Saccharomyces cerevisiae strain. The cybernetic model proposed is a new one that considers the degradation of starch not only into glucose but also into dextrins (reducing sugars) and takes into account the intracellular reactions occurring inside the cells, giving a more detailed description of the process. Furthermore, an identification procedure based on the Metropolis Monte Carlo optimization method coupled with a sensitivity analysis is proposed for the identification of the model's parameters, employing experimental data reported in the literature.

Enzymatic saccharification and bioethanol production from Cynara cardunculus pretreated by steam explosion.

Science.gov (United States)

Fernandes, Maria C; Ferro, Miguel D; Paulino, Ana F C; Mendes, Joana A S; Gravitis, Janis; Evtuguin, Dmitry V; Xavier, Ana M R B

2015-06-01

The correct choice of the specific lignocellulosic biomass pretreatment allows obtaining high biomass conversions for biorefinery implementations and cellulosic bioethanol production from renewable resources. Cynara cardunculus (cardoon) pretreated by steam explosion (SE) was involved in second-generation bioethanol production using separate hydrolysis and fermentation (SHF) or simultaneous saccharification and fermentation (SSF) processes. Steam explosion pretreatment led to partial solubilisation of hemicelluloses and increased the accessibility of residual polysaccharides towards enzymatic hydrolysis revealing 64% of sugars yield against 11% from untreated plant material. Alkaline extraction after SE pretreatment of cardoon (CSEOH) promoted partial removal of degraded lignin, tannins, extractives and hemicelluloses thus allowing to double glucose concentration upon saccharification step. Bioethanol fermentation in SSF mode was faster than SHF process providing the best results: ethanol concentration 18.7 g L(-1), fermentation efficiency of 66.6% and a yield of 26.6g ethanol/100 g CSEOH or 10.1 g ethanol/100 g untreated cardoon. Copyright © 2015 Elsevier Ltd. All rights reserved.

Optimization of Pretreatment and Enzymatic Saccharification of Cogon Grass Prior Ethanol Production

OpenAIRE

Jhalique Jane R. Fojas; Ernesto J. Del Rosario

2013-01-01

The dilute acid pretreatment and enzymatic saccharification of lignocellulosic substrate, cogon grass (Imperata cylindrical, L.) was optimized prior ethanol fermentation using simultaneous saccharification and fermentation (SSF) method. The optimum pretreatment conditions, temperature, sulfuric acid concentration, and reaction time were evaluated by determining the maximum sugar yield at constant enzyme loading. Cogon grass, at 10% w/v substrate loading, has optimum pretr...

Optimization of sodium hydroxide pretreatment and enzyme loading for efficient hydrolysis of rice straw to improve succinate production by metabolically engineered Escherichia coli KJ122 under simultaneous saccharification and fermentation.

Science.gov (United States)

Sawisit, Apichai; Jampatesh, Surawee; Jantama, Sirima Suvarnakuta; Jantama, Kaemwich

2018-07-01

Rice straw was pretreated with sodium hydroxide (NaOH) before subsequent use for succinate production by Escherichia coli KJ122 under simultaneous saccharification and fermentation (SSF). The NaOH pretreated rice straw was significantly enhanced lignin removal up to 95%. With the optimized enzyme loading of 4% cellulase complex + 0.5% xylanase (endo-glucanase 67 CMC-U/g, β-glucosidase 26 pNG-U/g and xylanase 18 CMC-U/g dry biomass), total sugar conversion reached 91.7 ± 0.8% (w/w). The physicochemical analysis of NaOH pretreated rice straw indicated dramatical changes in its structure, thereby favoring enzymatic saccharification. In batch SSF, succinate production of 69.8 ± 0.3 g/L with yield and productivity of 0.84 g/g pretreated rice straw and 0.76 ± 0.02 g/L/h, respectively, was obtained. Fed-batch SSF significantly improved succinate concentration and productivity to 103.1 ± 0.4 g/L and 1.37 ± 0.07 g/L/h with a comparable yield. The results demonstrated a feasibility of sequential saccharification and fermentation of rice straw as a promising process for succinate production in industrial scale. Copyright © 2018 Elsevier Ltd. All rights reserved.

Bioethanol Production From Banana Stem By Using Simultaneous Saccharification and Fermentation (SSF)

Science.gov (United States)

Kusmiyati; Mustofa, A.; Jumarmi

2018-05-01

The rapid growth and development of industries in the world result in a greater energy needs. Some studies show that ethanol can be used as an alternative energy. However, bioethanol production from food raw materials such as sugar and starch has drawback that cause the food crisis. This aim of this study was to convert banana stem into bioethanol. Banana stem contained of 44.6% cellulose, 36.0% hemicellulose and 19.4% lignin. After banana stems were pretreated with acid (H2SO4) and alkaline (NaOH) at a concentration of 2% w/v at 121 °C for 30 minutes, then subsequently the simultaneous saccharification and fermentation (SSF) were carried out by using mixed cultures of Aspergillus niger, Trichoderma reesei and Zymomonas mobilis at various enzymes ratios of (1:1:1), (1:2:1), (1:2:2), (1:1:2) and various pH (4, 5 and 6) with SSF time for 144 hours and temperature of 30°C. The results show that acid pretreatment showed better results than the alkali pretreatment. After acid pretreatment and alkali pretreatment, lignin content of pretreted banana stem reduced to 15.92% and 16.34%, respectively, cellulose increased to 52.11% and 50.6% respectively, hemicellulose reduced to 28.45% and 28.83%, respectively The SSF showed that pH 5 gave the highest bioethanol. The highest concentration of bioethanol (8.51 g/L) was achieved at the SSF process at pH 5 with a ratio Aspergillus niger, Trichoderma reesei and Zymomonas mobilis enzymes of (1:1:2).

Mechanistic insight into ultrasound induced enhancement of simultaneous saccharification and fermentation of Parthenium hysterophorus for ethanol production.

Science.gov (United States)

Singh, Shuchi; Agarwal, Mayank; Sarma, Shyamali; Goyal, Arun; Moholkar, Vijayanand S

2015-09-01

This paper presents investigations into mechanism of ultrasound assisted bioethanol synthesis using Parthenium hysterophorus biomass through simultaneous saccharification and fermentation (SSF) mode. Approach of coupling experimental results to mathematical model for SSF using Genetic Algorithm based optimization has been adopted. Comparison of model parameters for experiments with mechanical shaking and sonication (10% duty cycle) give an interesting mechanistic account of influence of ultrasound on SSF system. A 4-fold rise in ethanol and cell mass productivity is seen with ultrasound. The analysis reveals following facets of influence of ultrasound on SSF: increase in Monod constant for glucose for cell growth, maximal specific growth rate and inhibition constant of cell growth by glucose and reduction in specific cell death rate. Values of inhibition constant of cell growth by ethanol (K3E), and constants for growth associated (a) and non-growth associated (b) ethanol production remained unaltered with sonication. Beneficial effects of ultrasound are attributed to enhanced cellulose hydrolysis, enhanced trans-membrane transport of substrate and products as well as dilution of the toxic substances due to micro-convection induced by ultrasound. Intrinsic physiological functioning of cells remained unaffected by ultrasound as indicated by unaltered values of K3E, a and b. Copyright © 2015 Elsevier B.V. All rights reserved.

Dual effect of soluble materials in pretreated lignocellulose on simultaneous saccharification and co-fermentation process for the bioethanol production.

Science.gov (United States)

Qin, Lei; Li, Xia; Liu, Li; Zhu, Jia-Qing; Guan, Qi-Man; Zhang, Man-Tong; Li, Wen-Chao; Li, Bing-Zhi; Yuan, Ying-Jin

2017-01-01

In this study, wash liquors isolated from ethylenediamine and dry dilute acid pretreated corn stover were used to evaluate the effect of soluble materials in pretreated biomass on simultaneous saccharification and co-fermentation (SSCF) for ethanol production, respectively. Both of the wash liquors had different impacts on enzymatic hydrolysis and fermentation. Enzymatic conversions of glucan and xylan monotonically decreased as wash liquor concentration increased. Whereas, with low wash liquor concentrations, xylose consumption rate, cell viability and ethanol yield were maximally stimulated in fermentation without nutrient supplementary. Soluble lignins were found as the key composition which promoted sugars utilization and cell viability without nutrient supplementary. The dual effects of soluble materials on enzymatic hydrolysis and fermentation resulted in the reduction of ethanol yield as soluble materials increased in SSCF. Copyright © 2016 Elsevier Ltd. All rights reserved.

Dynamic modeling and analyses of simultaneous saccharification and fermentation process to produce bio-ethanol from rice straw.

Science.gov (United States)

Ko, Jordon; Su, Wen-Jun; Chien, I-Lung; Chang, Der-Ming; Chou, Sheng-Hsin; Zhan, Rui-Yu

2010-02-01

The rice straw, an agricultural waste from Asians' main provision, was collected as feedstock to convert cellulose into ethanol through the enzymatic hydrolysis and followed by the fermentation process. When the two process steps are performed sequentially, it is referred to as separate hydrolysis and fermentation (SHF). The steps can also be performed simultaneously, i.e., simultaneous saccharification and fermentation (SSF). In this research, the kinetic model parameters of the cellulose saccharification process step using the rice straw as feedstock is obtained from real experimental data of cellulase hydrolysis. Furthermore, this model can be combined with a fermentation model at high glucose and ethanol concentrations to form a SSF model. The fermentation model is based on cybernetic approach from a paper in the literature with an extension of including both the glucose and ethanol inhibition terms to approach more to the actual plants. Dynamic effects of the operating variables in the enzymatic hydrolysis and the fermentation models will be analyzed. The operation of the SSF process will be compared to the SHF process. It is shown that the SSF process is better in reducing the processing time when the product (ethanol) concentration is high. The means to improve the productivity of the overall SSF process, by properly using aeration during the batch operation will also be discussed.

Simultaneous or separated; comparison approach for saccharification and fermentation process in producing bio-ethanol from EFB

Science.gov (United States)

Bardant, Teuku Beuna; Dahnum, Deliana; Amaliyah, Nur

2017-11-01

Simultaneous Saccharification Fermentation (SSF) of palm oil (Elaeis guineensis) empty fruit bunch (EFB) pulp were investigated as a part of ethanol production process. SSF was investigated by observing the effect of substrate loading variation in range 10-20%w, cellulase loading 5-30 FPU/gr substrate and yeast addition 1-2%v to the ethanol yield. Mathematical model for describing the effects of these three variables to the ethanol yield were developed using Response Surface Methodology-Cheminformatics (RSM-CI). The model gave acceptable accuracy in predicting ethanol yield for Simultaneous Saccharification and Fermentation (SSF) with coefficient of determination (R2) 0.8899. Model validation based on data from previous study gave (R2) 0.7942 which was acceptable for using this model for trend prediction analysis. Trend prediction analysis based on model prediction yield showed that SSF gave trend for higher yield when the process was operated in high enzyme concentration and low substrate concentration. On the other hand, even SHF model showed better yield will be obtained if operated in lower substrate concentration, it still possible to operate in higher substrate concentration with slightly lower yield. Opportunity provided by SHF to operate in high loading substrate make it preferable option for application in commercial scale.

Direct Ethanol Production from Breadfruit Starch (Artocarpus communis Forst. by Engineered Simultaneous Saccharification and Fermentation (ESSF using Microbes Consortium

Directory of Open Access Journals (Sweden)

Iftachul Farida

2015-02-01

Full Text Available Breadfruit (Artocarpus communis Forst. is one of sources for ethanol production, which has high starch content (89%. Ethanol production from breadfruit starch was conducted by Simultaneous Saccharification and Fermentation (SSF technology using microbes consortium. The aim of the research was to examine a method to produce ethanol by SSF technology using microbes consortium at high yield and efficiency. The main research consisted of two treatments, namely normal SSF and enginereed SSF. The results showed that normal SSF using aeration and agitation during cultivation could produce ethanol at 11.15 ± 0.18 g/L, with the yield of product (Yp/s 0.34 g ethanol/g substrate; and yield of biomass (Yx/s 0.29 g cell/g substrate, respectively. A better result was obtained using engineered SSF in which aeration was stopped after biomass condition has reached the end of the exponential phase. The ethanol produced was 12.75 ± 0.04 g/L, with the yields of product (Yp/s 0.41 g ethanol/g substrate, and the yield of cell (Yx/s 0.09 g cell/g substrate.

Characterization of very high gravity ethanol fermentation of corn mash. Effect of glucoamylase dosage, pre-saccharification and yeast strain

DEFF Research Database (Denmark)

Devantier, Rasmus; Pedersen, S; Olsson, Lisbeth

2005-01-01

Ethanol was produced from very high gravity mashes of dry milled corn (35% w/w total dry matter) under simultaneous saccharification and fermentation conditions. The effects of glucoamylase dosage, pre-saccharification and Saccharomyces cerevisiae strain on the growth characteristics such as the ......Ethanol was produced from very high gravity mashes of dry milled corn (35% w/w total dry matter) under simultaneous saccharification and fermentation conditions. The effects of glucoamylase dosage, pre-saccharification and Saccharomyces cerevisiae strain on the growth characteristics...... such as the ethanol yield and volumetric and specific productivity were determined. It was shown that higher glucoamylase doses and/or pre-saccharification accelerated the simultaneous saccharification and fermentation process and increased the final ethanol concentration from 106 to 126 g/kg although the maximal...... specific growth rate was decreased. Ethanol production was not only growth related, as more than half of the total saccharides were consumed and more than half of the ethanol was produced during the stationary phase. Furthermore, a high stress tolerance of the applied yeast strain was found to be crucial...

Butanol production from lignocellulose by simultaneous fermentation, saccharification, and pervaporation or vacuum evaporation.

Science.gov (United States)

Díaz, Víctor Hugo Grisales; Tost, Gerard Olivar

2016-10-01

Techno-economic study of acetone, butanol and ethanol (ABE) fermentation from lignocellulose was performed. Simultaneous saccharification, fermentation and vacuum evaporation (SFS-V) or pervaporation (SFS-P) were proposed. A kinetic model of metabolic pathways for ABE fermentation with the effect of phenolics and furans in the growth was proposed based on published laboratory results. The processes were optimized in Matlab®. The end ABE purification was carried out by heat-integrated distillation. The objective function of the minimization was the total annualized cost (TAC). Fuel consumption of SFS-P using poly[1-(trimethylsilyl)-1-propyne] membrane was between 13.8 and 19.6% lower than SFS-V. Recovery of furans and phenolics for the hybrid reactors was difficult for its high boiling point. TAC of SFS-P was increased 1.9 times with supplementation of phenolics and furans to 3g/l each one for its high toxicity. Therefore, an additional detoxification method or an efficient pretreatment process will be necessary. Copyright © 2016 Elsevier Ltd. All rights reserved.

Ethanol production in a simultaneous saccharification and fermentation process with interconnected reactors employing hydrodynamic cavitation-pretreated sugarcane bagasse as raw material.

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Terán Hilares, Ruly; Ienny, João Vitor; Marcelino, Paulo Franco; Ahmed, Muhammad Ajaz; Antunes, Felipe A F; da Silva, Silvio Silvério; Santos, Júlio César Dos

2017-11-01

In this study, sugarcane bagasse (SCB) pretreated with alkali assisted hydrodynamic cavitation (HC) was investigated for simultaneous saccharification and fermentation (SSF) process for bioethanol production in interconnected column reactors using immobilized Scheffersomyces stipitis NRRL-Y7124. Initially, HC was employed for the evaluation of the reagent used in alkaline pretreatment. Alkalis (NaOH, KOH, Na 2 CO 3 , Ca(OH) 2 ) and NaOH recycled black liquor (successive batches) were used and their pretreatment effectiveness was assessed considering the solid composition and its enzymatic digestibility. In SSF process using NaOH-HC pretreatment SCB, 62.33% of total carbohydrate fractions were hydrolyzed and 17.26g/L of ethanol production (0.48g of ethanol/g of glucose and xylose consumed) was achieved. This proposed scheme of HC-assisted NaOH pretreatment together with our interconnected column reactors showed to be an interesting new approach for biorefineries. Copyright © 2017 Elsevier Ltd. All rights reserved.

L-lactic acid production from starch by simultaneous saccharification and fermentation in a genetically engineered Aspergillus oryzae pure culture.

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Wakai, Satoshi; Yoshie, Toshihide; Asai-Nakashima, Nanami; Yamada, Ryosuke; Ogino, Chiaki; Tsutsumi, Hiroko; Hata, Yoji; Kondo, Akihiko

2014-12-01

Lactic acid is a commodity chemical that can be produced biologically. Lactic acid-producing Aspergillus oryzae strains were constructed by genetic engineering. The A. oryzae LDH strain with the bovine L-lactate dehydrogenase gene produced 38 g/L of lactate from 100g/L of glucose. Disruption of the wild-type lactate dehydrogenase gene in A. oryzae LDH improved lactate production. The resulting strain A. oryzae LDHΔ871 produced 49 g/L of lactate from 100g/L of glucose. Because A. oryzae strains innately secrete amylases, A. oryzae LDHΔ871 produced approximately 30 g/L of lactate from various starches, dextrin, or maltose (all at 100 g/L). To our knowledge, this is the first report describing the simultaneous saccharification and fermentation of lactate from starch using a pure culture of transgenic A. oryzae. Our results indicate that A. oryzae could be a promising host for the bioproduction of useful compounds such as lactic acid. Copyright © 2014 Elsevier Ltd. All rights reserved.

Caffeic acid production by simultaneous saccharification and fermentation of kraft pulp using recombinant Escherichia coli.

Science.gov (United States)

Kawaguchi, Hideo; Katsuyama, Yohei; Danyao, Du; Kahar, Prihardi; Nakamura-Tsuruta, Sachiko; Teramura, Hiroshi; Wakai, Keiko; Yoshihara, Kumiko; Minami, Hiromichi; Ogino, Chiaki; Ohnishi, Yasuo; Kondo, Ahikiko

2017-07-01

Caffeic acid (3,4-dihydroxycinnamic acid) serves as a building block for thermoplastics and a precursor for biologically active compounds and was recently produced from glucose by microbial fermentation. To produce caffeic acid from inedible cellulose, separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) reactions were compared using kraft pulp as lignocellulosic feedstock. Here, a tyrosine-overproducing Escherichia coli strain was metabolically engineered to produce caffeic acid from glucose by introducing the genes encoding a 4-hydroxyphenyllactate 3-hydroxylase (hpaBC) from Pseudomonas aeruginosa and tyrosine ammonia lyase (fevV) from Streptomyces sp. WK-5344. Using the resulting recombinant strain, the maximum yield of caffeic acid in SSF (233 mg/L) far exceeded that by SHF (37.9 mg/L). In the SSF with low cellulase loads (≤2.5 filter paper unit/g glucan), caffeic acid production was markedly increased, while almost no glucose accumulation was detected, indicating that the E. coli cells experienced glucose limitation in this culture condition. Caffeic acid yield was also negatively correlated with the glucose concentration in the fermentation medium. In SHF, the formation of by-product acetate and the accumulation of potential fermentation inhibitors increased significantly with kraft pulp hydrolysate than filter paper hydrolysate. The combination of these inhibitors had synergistic effects on caffeic acid fermentation at low concentrations. With lower loads of cellulase in SSF, less potential fermentation inhibitors (furfural, 5-hydroxymethyfurfural, and 4-hydroxylbenzoic acid) accumulated in the medium. These observations suggest that glucose limitation in SSF is crucial for improving caffeic acid yield, owing to reduced by-product formation and fermentation inhibitor accumulation.

Periodic peristalsis increasing acetone-butanol-ethanol productivity during simultaneous saccharification and fermentation of steam-exploded corn straw.

Science.gov (United States)

Li, Jingwen; Wang, Lan; Chen, Hongzhang

2016-11-01

The acetone-butanol-ethanol (ABE) fermentation of lignocellulose at high solids content has recently attracted extensive attention. However, the productivity of high solids ABE fermentation of lignocellulose is typically low in traditional processes due to the lack of efficient intensifying methods. In the present study, periodic peristalsis, a novel intensifying method, was applied to improve ABE production by the simultaneous saccharification and fermentation (SSF) of steam-exploded corn straw using Clostridium acetobutylicum ATCC824. The ABE concentration and the ABE productivity of SSF at a solids content of 17.5% (w/w) with periodic peristalsis were 17.1 g/L and 0.20 g/(L h), respectively, which were higher than those obtained under static conditions (15.2 g/L and 0.14 g/(L h)). The initial sugar conversion rate over the first 12 h with periodic peristalsis was 4.67 g/(L h) at 10 FPU/g cellulase dosage and 15% (w/w) solids content, an increase of 49.7% compared with the static conditions. With periodic peristalsis, the period of batch fermentation was shortened from 108 h to 84 h. The optimal operating regime was a low frequency (6 h -1 ) of periodic peristalsis in the acid-production phase (0-48 h) of SSF. Therefore, periodic peristalsis should be an effective intensifying method to increase the productivity of ABE fermentation at high solids content. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Simultaneous saccharification and ethanol fermentation of oxalic acid pretreated corncob assessed with response surface methodology

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Jae-Won Lee; Rita C.L.B. Rodrigues; Thomas W. Jeffries

2009-01-01

Response surface methodology was used to evaluate optimal time, temperature and oxalic acid concentration for simultaneous saccharification and fermentation (SSF) of corncob particles by Pichia stipitis CBS 6054. Fifteen different conditions for pretreatment were examined in a 23 full factorial design with six axial points. Temperatures ranged from 132 to 180º...

Optimization of prehydrolysis time and substrate feeding to improve ethanol production by simultaneous saccharification and fermentation of furfural process residue.

Science.gov (United States)

He, Jianlong; Zhang, Wenbo; Liu, Xiaoyan; Xu, Ning; Xiong, Peng

2016-11-01

Ethanol is a very important industrial chemical. In order to improve ethanol productivity using Saccharomyces cerevisiae in fermentation from furfural process residue, we developed a process of simultaneous saccharification and fermentation (SSF) of furfural process residue, optimizing prehydrolysis cellulase loading concentration, prehydrolysis time, and substrate feeding strategy. The ethanol concentration obtained from the optimized process was 19.3 g/L, corresponding 76.5% ethanol yield, achieved by running SSF for 48 h from 10% furfural process residue with prehydrolysis at 50°C for 4 h and cellulase loading of 15 FPU/g furfural process residue. For higher ethanol concentrations, fed-batch fermentation was performed. The optimized fed-batch process increased the ethanol concentration to 37.6 g/L, 74.5% yield, obtained from 10% furfural process residue with two additions of 5% substrate at 12 and 24 h. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Simultaneous saccharification and cofermentation of lignocellulosic residues from commercial furfural production and corn kernels using different nutrient media

Directory of Open Access Journals (Sweden)

Cristhian Carrasco

2011-07-01

Full Text Available Abstract Background As the supply of starch grain and sugar cane, currently the main feedstocks for bioethanol production, become limited, lignocelluloses will be sought as alternative materials for bioethanol production. Production of cellulosic ethanol is still cost-inefficient because of the low final ethanol concentration and the addition of nutrients. We report the use of simultaneous saccharification and cofermentation (SSCF of lignocellulosic residues from commercial furfural production (furfural residue, FR and corn kernels to compare different nutritional media. The final ethanol concentration, yield, number of live yeast cells, and yeast-cell death ratio were investigated to evaluate the effectiveness of integrating cellulosic and starch ethanol. Results Both the ethanol yield and number of live yeast cells increased with increasing corn-kernel concentration, whereas the yeast-cell death ratio decreased in SSCF of FR and corn kernels. An ethanol concentration of 73.1 g/L at 120 h, which corresponded to a 101.1% ethanol yield based on FR cellulose and corn starch, was obtained in SSCF of 7.5% FR and 14.5% corn kernels with mineral-salt medium. SSCF could simultaneously convert cellulose into ethanol from both corn kernels and FR, and SSCF ethanol yield was similar between the organic and mineral-salt media. Conclusions Starch ethanol promotes cellulosic ethanol by providing important nutrients for fermentative organisms, and in turn cellulosic ethanol promotes starch ethanol by providing cellulosic enzymes that convert the cellulosic polysaccharides in starch materials into additional ethanol. It is feasible to produce ethanol in SSCF of FR and corn kernels with mineral-salt medium. It would be cost-efficient to produce ethanol in SSCF of high concentrations of water-insoluble solids of lignocellulosic materials and corn kernels. Compared with prehydrolysis and fed-batch strategy using lignocellulosic materials, addition of starch

Process analysis and optimization of simultaneous saccharification and co-fermentation of ethylenediamine-pretreated corn stover for ethanol production.

Science.gov (United States)

Qin, Lei; Zhao, Xiong; Li, Wen-Chao; Zhu, Jia-Qing; Liu, Li; Li, Bing-Zhi; Yuan, Ying-Jin

2018-01-01

Improving ethanol concentration and reducing enzyme dosage are main challenges in bioethanol refinery from lignocellulosic biomass. Ethylenediamine (EDA) pretreatment is a novel method to improve enzymatic digestibility of lignocellulose. In this study, simultaneous saccharification and co-fermentation (SSCF) process using EDA-pretreated corn stover was analyzed and optimized to verify the constraint factors on ethanol production. Highest ethanol concentration was achieved with the following optimized SSCF conditions at 6% glucan loading: 12-h pre-hydrolysis, 34 °C, pH 5.4, and inoculum size of 5 g dry cell/L. As glucan loading increased from 6 to 9%, ethanol concentration increased from 33.8 to 48.0 g/L, while ethanol yield reduced by 7%. Mass balance of SSCF showed that the reduction of ethanol yield with the increasing solid loading was mainly due to the decrease of glucan enzymatic conversion and xylose metabolism of the strain. Tween 20 and BSA increased ethanol concentration through enhancing enzymatic efficiency. The solid-recycled SSCF process reduced enzyme dosage by 40% (from 20 to 12 mg protein/g glucan) to achieve the similar ethanol concentration (~ 40 g/L) comparing to conventional SSCF. Here, we established an efficient SSCF procedure using EDA-pretreated biomass. Glucose enzymatic yield and yeast viability were regarded as the key factors affecting ethanol production at high solid loading. The extensive analysis of SSCF would be constructive to overcome the bottlenecks and improve ethanol production in cellulosic ethanol refinery.

Simultaneous saccharification and fermentation (SSF) of Jatropha curcas shells: utilization of co-products from the biodiesel production process.

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Visser, Evan Michael; Oliveira Filho, Delly; Tótola, Marcos Rogério; Martins, Marcio Arêdes; Guimarães, Valéria Monteze

2012-06-01

Jatropha curcas has great potential as an oil crop for use in biodiesel applications, and the outer shell is rich in lignocellulose that may be converted to ethanol, giving rise to the concept of a biorefinery. In this study, two dilute pretreatments of 0.5% H(2)SO(4) and 1.0% NaOH were performed on Jatropha shells with subsequent simultaneous saccharification and fermentation (SSF) of the pretreated water-insoluble solids (WIS) to evaluate the effect of inhibitors in the pretreatment slurry. A cellulase loading of 15 FPU/g WIS, complimented with an excess of cellobiase (19.25 U/g), was used for SSF of either the washed WIS or the original slurry to determine the effect of inhibitors. Ethanol and glucose were monitored during SSF of 20 g of pretreated biomass. The unwashed slurry showed to have a positive effect on SSF efficiency for the NaOH-pretreated biomass. Maximum efficiencies of glucan conversion to ethanol in the WIS were 40.43% and 41.03% for the H(2)SO(4)- and NaOH-pretreated biomasses, respectively.

Phenotypic selection of a wild Saccharomyces cerevisiae strain for simultaneous saccharification and co-fermentation of AFEX pretreated corn stover

Science.gov (United States)

Mingie Jin; Cory Sarks; Christa Gunawan; Benjamin D. Bice; Shane P. Simonett; Ragothaman Avanasi Narasimhan; Laura B. Willis; Bruce E. Dale; Venkatesh Balan; Trey K. Sato

2013-01-01

Simultaneous saccharification and co-fermentation (SSCF) process involves enzymatic hydrolysis of pretreated lignocellulosic biomass and fermentation of glucose and xylose in one bioreactor. The optimal temperatures for enzymatic hydrolysis are higher than the standard fermentation temperature of ethanologenic Saccharomyces cerevisiae. Moreover,...

High solid simultaneous saccharification and fermentation of wet oxidized corn stover to ethanol

DEFF Research Database (Denmark)

Varga, E.; Klinke, H.B.; Reczey, K.

2004-01-01

In this study ethanol was produced from corn stover pretreated by alkaline and acidic wet oxidation (WO) (195 degreesC, 15 min, 12 bar oxygen) followed by nonisothermal simultaneous saccharification and fermentation (SSF). In the first step of the SSF, small amounts of cellulases were added at 50...... increase of substrate concentration reduced the ethanol yield significant as a result of insufficient mass transfer. It was also shown that the fermentation could be followed with an easy monitoring system based on the weight loss of the produced CO2. (C) 2004 Wiley Periodicals, Inc....

Valorization of kitchen biowaste for ethanol production via simultaneous saccharification and fermentation using co-cultures of the yeasts Saccharomyces cerevisiae and Pichia stipitis.

Science.gov (United States)

Ntaikou, Ioanna; Menis, Nikolaos; Alexandropoulou, Maria; Antonopoulou, Georgia; Lyberatos, Gerasimos

2018-04-30

The biotransformation of the pre-dried and shredded organic fraction of kitchen waste to ethanol was investigated, via co-cultures of the yeasts Saccharomyces cerevisiae and Pichia stipitis (Scheffersomyces stipitis). Preliminary experiments with synthetic media were performed, in order to investigate the effect of different operational parameters on the ethanol production efficiency of the co-culture. The control of the pH and the supplementation with organic nitrogen were shown to be key factors for the optimization of the process. Subsequently, the ethanol production efficiency from the waste was assessed via simultaneous saccharification and fermentation experiments. Different loadings of cellulolytic enzymes and mixtures of cellulolytic with amylolytic enzymatic blends were tested in order to enhance the substrate conversion efficiency. It was further shown that for solids loading up to 40% waste on dry mass basis, corresponding to 170 g.L -1 initial concentration of carbohydrates, no substrate inhibition occurred, and ethanol concentration up to 45 g.L -1 was achieved. Copyright © 2018 Elsevier Ltd. All rights reserved.

Enhanced Inulin Saccharification by Self-Produced Inulinase from a Newly Isolated Penicillium sp. and its Application in D-Lactic Acid Production.

Science.gov (United States)

Zheng, Zhaojuan; Xu, Qianqian; Liu, Peng; Zhou, Fan; Ouyang, Jia

2018-03-10

In order to find an alternative for commercial inulinase, a strain XL01 identified as Penicillium sp. was screened for inulinase production. The broth after cultivated was centrifuged, filtered, and used as crude enzyme for the following saccharification. At pH 5.0 and 50 °C, the crude enzyme released 84.9 g/L fructose and 20.7 g/L glucose from 120 g/L inulin in 72 h. In addition, simultaneous saccharification and fermentation of chicory flour for D-lactic acid production was carried out using the self-produced crude inulinase and Lactobacillus bulgaricus CGMCC 1.6970. A high D-lactic acid titer and productivity of 122.0 g/L and 1.69 g/(L h) was achieved from 120 g/L chicory flour in 72 h. The simplicity for inulinase production and the high efficiency for D-lactic acid fermentation provide a perspective and profitable industrial biotechnology for utilization of the inulin-rich biomass.

Evaluation of Ethanol Production Activity by Engineered Saccharomyces cerevisiae Fermenting Cellobiose through the Phosphorolytic Pathway in Simultaneous Saccharification and Fermentation of Cellulose.

Science.gov (United States)

Lee, Won-Heong; Jin, Yong-Su

2017-09-28

In simultaneous saccharification and fermentation (SSF) for production of cellulosic biofuels, engineered Saccharomyces cerevisiae capable of fermenting cellobiose has provided several benefits, such as lower enzyme costs and faster fermentation rate compared with wild-type S. cerevisiae fermenting glucose. In this study, the effects of an alternative intracellular cellobiose utilization pathway-a phosphorolytic pathway based on a mutant cellodextrin transporter (CDT-1 (F213L)) and cellobiose phosphorylase (SdCBP)-was investigated by comparing with a hydrolytic pathway based on the same transporter and an intracellular β-glucosidase (GH1-1) for their SSF performances under various conditions. Whereas the phosphorolytic and hydrolytic cellobiose-fermenting S. cerevisiae strains performed similarly under the anoxic SSF conditions, the hydrolytic S. cerevisiae performed slightly better than the phosphorolytic S. cerevisiae under the microaerobic SSF conditions. Nonetheless, the phosphorolytic S. cerevisiae expressing the mutant CDT-1 showed better ethanol production than the glucose-fermenting S. cerevisiae with an extracellular β-glucosidase, regardless of SSF conditions. These results clearly prove that introduction of the intracellular cellobiose metabolic pathway into yeast can be effective on cellulosic ethanol production in SSF. They also demonstrate that enhancement of cellobiose transport activity in engineered yeast is the most important factor affecting the efficiency of SSF of cellulose.

Characteristics of Corn Stover Pretreated with Liquid Hot Water and Fed-Batch Semi-Simultaneous Saccharification and Fermentation for Bioethanol Production

Science.gov (United States)

Li, Xuezhi; Lu, Jie; Zhao, Jian; Qu, Yinbo

2014-01-01

Corn stover is a promising feedstock for bioethanol production because of its abundant availability in China. To obtain higher ethanol concentration and higher ethanol yield, liquid hot water (LHW) pretreatment and fed-batch semi-simultaneous saccharification and fermentation (S-SSF) were used to enhance the enzymatic digestibility of corn stover and improve bioconversion of cellulose to ethanol. The results show that solid residues from LHW pretreatment of corn stover can be effectively converted into ethanol at severity factors ranging from 3.95 to 4.54, and the highest amount of xylan removed was approximately 89%. The ethanol concentrations of 38.4 g/L and 39.4 g/L as well as ethanol yields of 78.6% and 79.7% at severity factors of 3.95 and 4.54, respectively, were obtained by fed-batch S-SSF in an optimum conditions (initial substrate consistency of 10%, and 6.1% solid residues added into system at the prehydrolysis time of 6 h). The changes in surface morphological structure, specific surface area, pore volume and diameter of corn stover subjected to LHW process were also analyzed for interpreting the possible improvement mechanism. PMID:24763192

Comparison of high-titer lactic acid fermentation from NaOH- and NH3-H2O2-pretreated corncob by Bacillus coagulans using simultaneous saccharification and fermentation

Science.gov (United States)

Zhang, Zhenting; Xie, Yuejiao; He, Xiaolan; Li, Xinli; Hu, Jinlong; Ruan, Zhiyong; Zhao, Shumiao; Peng, Nan; Liang, Yunxiang

2016-01-01

Lignocellulose is one of the most abundant renewable feedstocks that has attracted considerable attention as a substrate for biofuel and biochemical production. One such biochemical product, lactic acid, is an important fermentation product because of its great potential for the production of biodegradable and biocompatible polylactic acid. High-titer lactic acid production from lignocellulosic materials has been achieved recently; however, it requires biodetoxification or results in large amounts of waste washing water. In this study, we employed two alkaline pretreatment methods and compared their effects on lactic acid fermentation of pretreated corncob by Bacillus coagulans LA204 using fed-batch simultaneous saccharification and fermentation under non-sterile conditions. The lactic acid titer, yield, and productivity from 16% (w/w) NaOH-pretreated and washed corncob were 122.99 g/L, 0.77 g/g corncob, and 1.37 g/L/h, respectively, and from 16% NH3-H2O2-pretreated and washed corncob were 118.60 g/L, 0.74 g/g corncob, and 1.32 g/L/h, respectively. Importantly, the lactic acid titer, yield, and productivity from 18.4% NH3-H2O2-pretreated and unwashed corncob by using fed-batch simultaneous saccharification and fermentation reached 79.47 g/L, 0.43 g/g corncob, and 1.10 g/L/h, respectively, demonstrating that this method is possible for industrial applications and saves washing water. PMID:27853308

Comparison of high-titer lactic acid fermentation from NaOH- and NH3-H2O2-pretreated corncob by Bacillus coagulans using simultaneous saccharification and fermentation.

Science.gov (United States)

Zhang, Zhenting; Xie, Yuejiao; He, Xiaolan; Li, Xinli; Hu, Jinlong; Ruan, Zhiyong; Zhao, Shumiao; Peng, Nan; Liang, Yunxiang

2016-11-17

Lignocellulose is one of the most abundant renewable feedstocks that has attracted considerable attention as a substrate for biofuel and biochemical production. One such biochemical product, lactic acid, is an important fermentation product because of its great potential for the production of biodegradable and biocompatible polylactic acid. High-titer lactic acid production from lignocellulosic materials has been achieved recently; however, it requires biodetoxification or results in large amounts of waste washing water. In this study, we employed two alkaline pretreatment methods and compared their effects on lactic acid fermentation of pretreated corncob by Bacillus coagulans LA204 using fed-batch simultaneous saccharification and fermentation under non-sterile conditions. The lactic acid titer, yield, and productivity from 16% (w/w) NaOH-pretreated and washed corncob were 122.99 g/L, 0.77 g/g corncob, and 1.37 g/L/h, respectively, and from 16% NH 3 -H 2 O 2 -pretreated and washed corncob were 118.60 g/L, 0.74 g/g corncob, and 1.32 g/L/h, respectively. Importantly, the lactic acid titer, yield, and productivity from 18.4% NH 3 -H 2 O 2 -pretreated and unwashed corncob by using fed-batch simultaneous saccharification and fermentation reached 79.47 g/L, 0.43 g/g corncob, and 1.10 g/L/h, respectively, demonstrating that this method is possible for industrial applications and saves washing water.

Characterization of very high gravity ethanol fermentation of corn mash. Effect of glucoamylase dosage, pre-saccharification and yeast strain

Energy Technology Data Exchange (ETDEWEB)

Devantier, R. [Starch, Applied Discovery, Research and Development, Novozymes A/S, Bagsvaerd (Denmark); Center for Microbial Biotechnology, BioCentrum-DTU, Technical Univ. of Denmark, Kgs Lyngby (Denmark); Pedersen, S. [Starch, Applied Discovery, Research and Development, Novozymes A/S, Bagsvaerd (Denmark); Olsson, L. [Center for Microbial Biotechnology, BioCentrum-DTU, Technical Univ. of Denmark, Kgs Lyngby (Denmark)

2005-09-01

Ethanol was produced from very high gravity mashes of dry milled corn (35% w/w total dry matter) under simultaneous saccharification and fermentation conditions. The effects of glucoamylase dosage, pre-saccharification and Saccharomyces cerevisiae strain on the growth characteristics such as the ethanol yield and volumetric and specific productivity were determined. It was shown that higher glucoamylase doses and/or pre-saccharification accelerated the simultaneous saccharification and fermentation process and increased the final ethanol concentration from 106 to 126 g/kg although the maximal specific growth rate was decreased. Ethanol production was not only growth related, as more than half of the total saccharides were consumed and more than half of the ethanol was produced during the stationary phase. Furthermore, a high stress tolerance of the applied yeast strain was found to be crucial for the outcome of the fermentation process, both with regard to residual saccharides and final ethanol concentration. The increased formation of cell mass when a well-suited strain was applied increased the final ethanol concentration, since a more complete fermentation was achieved. (orig.)

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

Energy Technology Data Exchange (ETDEWEB)

Sukumaran, Rajeev K.; Singhania, Reeta Rani; Mathew, Gincy Marina; Pandey, Ashok [Biotechnology Division, National Institute for Interdisciplinary Science and Technology, CSIR, Trivandrum-695 019 (India)

2009-02-15

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 {beta}-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. (author)

Evaluation of Potential Fungal Species for the in situ Simultaneous Saccharification and Fermentation (SSF of Cellulosic Material

Directory of Open Access Journals (Sweden)

Leeuwen, J.

2011-01-01

Full Text Available Three fungal species were evaluated for their abilities to saccharify pure cellulose. The three species chosen represented three major wood-rot molds; brown rot (Gloeophyllum trabeum, white rot (Phanerochaete chrysosporium and soft rot (Trichoderma reesei. After solid state fermentation of the fungi on the filter paper for four days, the saccharified cellulose was then fermented to ethanol by using Saccharomyces cerevisiae. The efficiency of the fungal species in saccharifying the filter paper was compared against a low dose (25 FPU/g cellulose of a commercial cellulase. Total sugar, cellobiose and glucose were monitored during the fermentation period, along with ethanol, acetic acid and lactic acid. Results indicated that the most efficient fungal species in saccharifying the filter paper was T. reesei with 5.13 g/100 g filter paper of ethanol being produced at days 5, followed by P. chrysosporium at 1.79 g/100 g filter paper. No ethanol was detected for the filter paper treated with G. trabeum throughout the five day fermentation stage. Acetic acid was only produced in the sample treated with T. reesei and the commercial enzyme, with concentration 0.95 and 2.57 g/100 g filter paper, respectively at day 5. Lactic acid production was not detected for all the fungal treated filter paper after day 5. Our study indicated that there is potential in utilizing in situ enzymatic saccharification of biomass by using T. reesei and P. chrysosporium that may lead to an economical simultaneous saccharification and fermentation process for the production of fuel ethanol.

Ethanol production from alfalfa fiber fractions by saccharification and fermentation

Energy Technology Data Exchange (ETDEWEB)

Sreenath, H.K. [University of Wisconsin, Madison, WI (United States). Dept. of Biological Systems Engineering; USDA Forest Service, Madison, WI (United States). Forest Products Lab.; Koegel, R.G. [US Department of Agriculture, Madison, WI (United States). Dairy Forage Research Center; Moldes, A.B. [USDA Forest Service, Madison, WI (United States). Forest Products Lab.; Universidade de Vigo, Ourense (Spain); Jeffries, T.W. [USDA Forest Service, Madison, WI (United States). Forest Products Lab.; Straub, R.J. [University of Wisconsin, Madison, WI (United States). Dept. of Biological Systems Engineering

2001-07-01

This work describes ethanol production from alfalfa fiber using separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) with and without liquid hot water (LHW) pretreatment. Candida shehatae FPL-702 produced 5 and 6.4 g/l ethanol with a yield of 0.25 and 0.16 g ethanol/g sugar respectively by SHF and SSF from alfalfa fiber without pretreatment. With LHW pretreatment using SSF, C. shehatae FPL-702 produced 18.0 g/l ethanol, a yield of 0.45 g ethanol/g sugar from cellulosic solids or 'raffinate'. Using SHF, it produced 9.6 g/l ethanol, a yield of 0.47 g ethanol/g sugar from raffinate. However, the soluble extract fraction containing hemicelluloses was poorly fermented in both SHF and SSF due to the presence of inhibitors. Addition of dilute acid during LHW pretreatment of alfalfa fiber resulted in fractions that were poorly saccharified and fermented. These results show that unpretreated alfalfa fiber produced a lower ethanol yield. Although LHW pretreatment can increase ethanol production from raffinate fiber fractions, it does not increase production from the hemicellulosic and pectin fractions. (author)

A Novel simultaneous-Saccharification-Fermentation Strategy for Efficient Co-fermentation of C5 and C6 Sugars Using Native, Non-GMO Yeasts

Energy Technology Data Exchange (ETDEWEB)

Varanasi, Sasidhar [Univ. of Toledo, OH (United States); Relue, Patricia [Univ. of Toledo, OH (United States)

2013-09-30

Economic bioethanol production is critically dependent upon the ability to convert both the hexose (C6) and pentose (C5) sugars resulting from cellulose and hemicellulose. C5 sugars are not readily fermentable by native Saccharomyces cerevisiae. Genetically Modified Organisms (GMOs) are designed to ferment xylose, but their stability, ethanol yield, environmental impact, and survival under conditions of industrial fermentation are unproven. In this project, we developed a novel approach for efficient fermentation of both C5 and C6 sugars using native S. Cerevisiae by exploiting its ability to produce ethanol from xylulose - the keto-isomer of xylose. While the isomerization of xylose to xylulose can be accomplished via commercially (and cheaply) available Xylose Isomerase (XI) (Sweetzyme™), this conversion has an extremely unfavorable equilibrium (xylose:xylose is about 5:1). To address this, we developed two alternate strategies. In the first, the two enzymes XI and urease are coimmobilized on solid support particles to enable complete isomerization of xylose to xylulose under pH conditions suitable for fermentation, in a simultaneous-isomerization-fermentation (SIF) mode. The ability of our technology to conduct isomerization of xylose under pH conditions suitable for both saccharification and fermentation opens the possibility of SSF with native yeasts for the first time. Herein, we performed specific research tasks for implementation of our technology in several modes of operation, including simultaneous-isomerization-and-fermentation (SIF), simultaneous-saccharification-and-isomerization (SSI) followed by fermentation, and SSF mode with the biomass feedstock poplar. The projected economics of our process are very favorable in comparison to the costs associated with engineering, licensing and propagating GMOs. This novel fermentation technology is readily accessible to rural farming economies for implementation in cellulosic ethanol production facilities.

Effect of technological parameters the process of ethanol production in the saccharification of barley

Directory of Open Access Journals (Sweden)

G. V. Agafonov

2016-01-01

Full Text Available Tenderized saccharification process is a mass of the cores in the production of ethanol from starch-containing raw materials. Saccharification of starchy raw materials needed because main production races alcohol yeast do not synthesize and amylolytic enzymes are not able to use in their metabolism Nepo-starch, facility and internalize only mono – and disaccharides. The main aim is to obtain a mash saccharification with a high content of fermentable carbohydrates. The main objective is to obtain a saccharification wort with a high con-zhaniem fermentable carbohydrates. From the course of the saccharification process, the quality of the alcohol depends on the completeness of the fermentation of starch raw materials and product yield. On saccharification sodden mass affected by temperature, pH, glucoamylase dosage and duration of the process. From these factors depend technical-economic production figures. Saccharification boiled barley weight carried glucoamylase enzyme preparation bios 800 A at the rate of 6.0 units per g starch SFS. Control is tenderized weight obtained using one enzyme preparation Alfaferm 3500 A at a dose of 1.0 units per g starch AS. Saccharification takes place at a temperature of 50–65 0 C and pH 4.0–5.5 for 120 minutes. pH was adjusted weight tenderized by making it in sulfuric acid. The saccharification process dynamics studied glucose accumulation in the barley wort in function of temperature, pH, and the dosage of glucoamylase. Samples for glucose determination were taken every 30 minutes. At a temperature of 65 0 C in the first 30 minutes of the accumulation of glucose goes fast, then suddenly slows down, it is possible due to the inactivation of glucoamylase due to changes in the structure of the enzyme molecule. The maximum amount of glucose stored at a temperature of 60 0 C and 11.1 g / cm3, which corresponds to the optimum action of glucoamylase enzyme preparation used. Theconcentration of

Saccharification and hydrolytic enzyme production of alkali pre-treated wheat bran by Trichoderma virens under solid state fermentation.

Science.gov (United States)

El-Shishtawy, Reda M; Mohamed, Saleh A; Asiri, Abdullah M; Gomaa, Abu-Bakr M; Ibrahim, Ibrahim H; Al-Talhi, Hasan A

2015-05-28

In continuation of our previously interest in the saccharification of agriculture wastes by Bacillus megatherium in solid state fermentation (SSF), we wish to report an investigation and comparative evaluation among Trichoderma sp. for the saccharification of four alkali-pretreated agricultural residues and production of hydrolytic enzymes, carboxymethyl cellulase (CMCase), filter paperase (FPase), pectinase (PGase) and xylanase (Xylase) in SSF. The optimization of the physiological conditions of production of hydrolytic enzymes and saccharification content from Trichoderma virens using alkali-pretreated wheat bran was the last goal. The physico-chemical parameters of SSF include incubation time, incubation temperature, moisture content of the substrate, incubation pH, supplementation with carbon and nitrogen sources were optimized. Saccharification of different solid state fermentation sources wheat bran, date's seeds, grass and palm leaves, were tested for the production of fermentable sugar by Trichoderma sp. The maximum production of hydrolytic enzymes CMCase, FPase, PGase and Xylase and saccharification content were obtained on wheat bran. Time course, moisture content, optimum temperature, optimum pH, supplementation with carbon and nitrogen sources were optimized to achieve the maximum production of the hydrolytic enzymes, protein and total carbohydrate of T. virens using alkali pre-treated wheat bran. The maximum production of CMCase, FPase, PGase, Xylase, protein and carbohydrate content was recorded at 72 h of incubation, 50-70 % moisture, temperature 25-35 °C and pH 5. The influence of supplementary carbon and nitrogen sources was studied. While lactose and sucrose enhanced the activity of PGase from 79.2 to 582.9 and 632.6 U/g, starch inhibited all other enzymes. This was confirmed by maximum saccharification content. Among the nitrogen sources, yeast extract and urea enhanced the saccharification content and CMCase, PGase and Xylase. The results of

Comparison of Pretreatment Methods on Vetiver Leaves for Efficient Processes of Simultaneous Saccharification and Fermentation by Neurospora sp.

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Restiawaty, E.; Dewi, A.

2017-07-01

Lignocellulosic biomass is a potential raw material for bioethanol production. Neurospora sp. can be used to convert lignocellulosic biomass into bioethanol because of its ability to perform simultaneous saccharification and fermentation. However, lignin content, degree of polymerization, and crystallinity of cellulose contained in lignocellulosic biomass can inhibit cellulosic-biomass digestion by Neurospora sp, so that a suitable pretreatment method of lignocellulosic biomass is needed. The focus of this research was to investigate the suitable pretreatment method for vetiver leaves (Vetiveria zizanioides L. Nash) used as a raw material producing bioethanol in the process of simultaneous saccharification and fermentation (SSF) by Neurospora sp.. Vetiver plants obtained from Garut are deliberately cultivated to produce essential oils extracted from the roots of this plant. Since the vetiver leaves do not contain oil, some of harvested leaves are usually used for crafts and cattle feed, and the rest are burned. This study intended to look at other potential of vetiver leaves as a source of renewable energy. Pretreatments of the vetiver leaves were conducted using hot water, dilute acid, alkaline & dilute acid, and alkaline peroxide, in which each method was accompanied by thermal treatment. The results showed that the alkaline peroxide treatment is a suitable for vetiver leaves as indicated by the increase of cellulose content up to 65.1%, while the contents of hot water soluble, hemicellulose, lignin, and ash are 8.7%, 18.3%, 6.8%, and 1.1%, respectively. Using this pretreatment method, the vetiver leaves can be converted into bioethanol by SSF process using Neurospora sp. with a concentration of bioethanol of 6.7 g/L operated at room temperature.

Modeling and experimental studies on intermittent starch feeding and citrate addition in simultaneous saccharification and fermentation of starch to flavor compounds.

Science.gov (United States)

Chavan, Abhijit R; Raghunathan, Anuradha; Venkatesh, K V

2009-04-01

Simultaneous saccharification and fermentation (SSF) is a combined process of saccharification of a renewable bioresource and fermentation process to produce products, such as lactic acid and ethanol. Recently, SSF has been extensively used to convert various sources of cellulose and starch into fermentative products. Here, we present a study on production of buttery flavors, namely diacetyl and acetoin, by growing Lactobacillus rhamnosus on a starch medium containing the enzyme glucoamylase. We further develop a structured kinetics for the SSF process, which includes enzyme and growth kinetics. The model was used to simulate the effect of pH and temperature on the SSF process so as to obtain optimum operating conditions. The model was experimentally verified by conducting SSF using an initial starch concentration of 100 g/L. The study demonstrated that the developed kinetic was able to suggest strategies for improved productivities. The developed model was able to accurately predict the enhanced productivity of flavors in a three stage process with intermittent addition of starch. Experimental and simulations demonstrated that citrate addition can also lead to enhanced productivity of flavors. The developed optimal model for SSF was able to capture the dynamics of SSF in batch mode as well as in a three stage process. The structured kinetics was also able to quantify the effect of multiple substrates present in the medium. The study demonstrated that structured kinetic models can be used in the future for design and optimization of SSF as a batch or a fed-batch process.

Efficient Simultaneous Saccharification and Fermentation of Inulin to 2,3-Butanediol by Thermophilic Bacillus licheniformis ATCC 14580

Science.gov (United States)

Li, Lixiang; Chen, Chao; Li, Kun; Wang, Yu; Gao, Chao; Ma, Cuiqing

2014-01-01

2,3-Butanediol (2,3-BD) is an important starting material for the manufacture of bulk chemicals. For efficient and large-scale production of 2,3-BD through fermentation, low-cost substrates are required. One such substrate, inulin, is a polydisperse fructan found in a wide variety of plants. In this study, a levanase with high inulinase activity and high pH and temperature stability was identified in Bacillus licheniformis strain ATCC 14580. B. licheniformis strain ATCC 14580 was found to efficiently produce 2,3-BD from fructose at 50°C. Then, the levanase was used for simultaneous saccharification and fermentation (SSF) of inulin to 2,3-BD. A fed-batch SSF yielded 103.0 g/liter 2,3-BD in 30 h, with a high productivity of 3.4 g/liter · h. The results suggest that the SSF process developed with the thermophilic B. licheniformis strain used might be a promising alternative for efficient 2,3-BD production from the favorable substrate inulin. PMID:25107977

High-titer lactic acid production from NaOH-pretreated corn stover by Bacillus coagulans LA204 using fed-batch simultaneous saccharification and fermentation under non-sterile condition.

Science.gov (United States)

Hu, Jinlong; Zhang, Zhenting; Lin, Yanxu; Zhao, Shumiao; Mei, Yuxia; Liang, Yunxiang; Peng, Nan

2015-04-01

Lactic acid (LA) is an important chemical with various industrial applications. Non-food feedstock is commercially attractive for use in LA production; however, efficient LA fermentation from lignocellulosic biomass resulting in both high yield and titer faces technical obstacles. In this study, the thermophilic bacterium Bacillus coagulans LA204 demonstrated considerable ability to ferment glucose, xylose, and cellobiose to LA. Importantly, LA204 produces LA from several NaOH-pretreated agro stovers, with remarkably high yields through simultaneous saccharification and fermentation (SSF). A fed-batch SSF process conducted at 50°C and pH 6.0, using a cellulase concentration of 30 FPU (filter paper unit)/g stover and 10 g/L yeast extract in a 5-L bioreactor, was developed to produce LA from 14.4% (w/w) NaOH-pretreated non-sterile corn stover. LA titer, yield, and average productivity reached 97.59 g/L, 0.68 g/g stover, and 1.63 g/L/h, respectively. This study presents a feasible process for lignocellulosic LA production from abundant agro stovers. Copyright © 2015 Elsevier Ltd. All rights reserved.

Model-based optimization and scale-up of multi-feed simultaneous saccharification and co-fermentation of steam pre-treated lignocellulose enables high gravity ethanol production.

Science.gov (United States)

Wang, Ruifei; Unrean, Pornkamol; Franzén, Carl Johan

2016-01-01

High content of water-insoluble solids (WIS) is required for simultaneous saccharification and co-fermentation (SSCF) operations to reach the high ethanol concentrations that meet the techno-economic requirements of industrial-scale production. The fundamental challenges of such processes are related to the high viscosity and inhibitor contents of the medium. Poor mass transfer and inhibition of the yeast lead to decreased ethanol yield, titre and productivity. In the present work, high-solid SSCF of pre-treated wheat straw was carried out by multi-feed SSCF which is a fed-batch process with additions of substrate, enzymes and cells, integrated with yeast propagation and adaptation on the pre-treatment liquor. The combined feeding strategies were systematically compared and optimized using experiments and simulations. For high-solid SSCF process of SO2-catalyzed steam pre-treated wheat straw, the boosted solubilisation of WIS achieved by having all enzyme loaded at the beginning of the process is crucial for increased rates of both enzymatic hydrolysis and SSCF. A kinetic model was adapted to simulate the release of sugars during separate hydrolysis as well as during SSCF. Feeding of solid substrate to reach the instantaneous WIS content of 13 % (w/w) was carried out when 60 % of the cellulose was hydrolysed, according to simulation results. With this approach, accumulated WIS additions reached more than 20 % (w/w) without encountering mixing problems in a standard bioreactor. Feeding fresh cells to the SSCF reactor maintained the fermentation activity, which otherwise ceased when the ethanol concentration reached 40-45 g L(-1). In lab scale, the optimized multi-feed SSCF produced 57 g L(-1) ethanol in 72 h. The process was reproducible and resulted in 52 g L(-1) ethanol in 10 m(3) scale at the SP Biorefinery Demo Plant. SSCF of WIS content up to 22 % (w/w) is reproducible and scalable with the multi-feed SSCF configuration and model-aided process

Simultaneous Saccharification and Fermentation of Lactic Acid from Empty Fruit Bunch at High Solids Loading

Directory of Open Access Journals (Sweden)

Nursia Hassan

2016-03-01

Full Text Available The production of value-added chemicals from the bioconversion of lignocellulose biomass has been considered a promising venture. In this study, microwave, alkali-pretreated empty fruit bunch (EFB was used as the substrate, utilizing pelletized filamentous Rhizopus oryzae NRRL 395 and cellulolytic enzymes for lactic acid production in a fed-batch simultaneous saccharification and fermentation (SSF process. Insoluble solids generally do not affect the SSF process until a certain concentration is exceeded. To achieve a high lactic acid concentration in the broth, a high solids loading was required to allow a higher rate of glucose conversion. However, the results revealed a decrease in the final lactic acid yield when running SSF at a massive insoluble solids level. High osmotic pressure in the medium led to poor cellular performance and caused the Rhizopus oryzae pellets to break down, affecting the lactic acid production. To improve the process performance, a fed-batch operation mode was used. The fed-batch operation was shown to facilitate higher lactic acid yield, compared with the SSF batch mode. Enzyme feeding, as well as substrate feeding, was also investigated as a means of enabling a higher dry matter content, with a high glucose conversion in SSF of cellulose-rich EFB.

SACCHARIFICATION WITH Phanerochaete chrysosporium and ...

African Journals Online (AJOL)

cantocanche

2011-05-09

May 9, 2011 ... and Pleurotus ostreatus enzymatic extracts of pretreated banana ... reduction in the cost of production (Hahn-Hägerdal et al., ... economy, the saccharification of these materials is necessary ... To efficiently process lignocellulosic wastes, the lignin ..... industries depend on the success in saccharification.

Simultaneous saccharification and aerobic fermentation of high titer cellulosic citric acid by filamentous fungus Aspergillus niger.

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Hou, Weiliang; Bao, Jie

2018-04-01

Simultaneous saccharification and fermentation (SSF) is the most efficient operation in biorefining conversion, but aerobic SSF under high solids loading significantly faces the serious oxygen transfer limitation. This study took the first insight into an aerobic SSF by high oxygen demanding filamentous fungi in highly viscous lignocellulose hydrolysate. The results show that oxygen requirement in the aerobic SSF by Aspergillus niger was well satisfied for production of cellulosic citric acid. The record high citric acid titer of 136.3 g/L and the overall conversion yield of 74.9% of cellulose were obtained by the aerobic SSF. The advantage of SSF to the separate hydrolysis and fermentation (SHF) on citric acid fermentation was compared based on the rigorous Aspen Plus modeling. The techno-economic analysis indicates that the minimum citric acid selling price (MCSP) of $0.603 per kilogram by SSF was highly competitive with the commercial citric acid from starch feedstock. Copyright © 2018 Elsevier Ltd. All rights reserved.

Enhanced production of raw starch degrading enzyme using agro-industrial waste mixtures by thermotolerant Rhizopus microsporus for raw cassava chip saccharification in ethanol production.

Science.gov (United States)

Trakarnpaiboon, Srisakul; Srisuk, Nantana; Piyachomkwan, Kuakoon; Sakai, Kenji; Kitpreechavanich, Vichien

2017-09-14

In the present study, solid-state fermentation for the production of raw starch degrading enzyme was investigated by thermotolerant Rhizopus microsporus TISTR 3531 using a combination of agro-industrial wastes as substrates. The obtained crude enzyme was applied for hydrolysis of raw cassava starch and chips at low temperature and subjected to nonsterile ethanol production using raw cassava chips. The agro-industrial waste ratio was optimized using a simplex axial mixture design. The results showed that the substrate mixture consisting of rice bran:corncob:cassava bagasse at 8 g:10 g:2 g yielded the highest enzyme production of 201.6 U/g dry solid. The optimized condition for solid-state fermentation was found as 65% initial moisture content, 35°C, initial pH of 6.0, and 5 × 10 6 spores/mL inoculum, which gave the highest enzyme activity of 389.5 U/g dry solid. The enzyme showed high efficiency on saccharification of raw cassava starch and chips with synergistic activities of commercial α-amylase at 50°C, which promotes low-temperature bioethanol production. A high ethanol concentration of 102.2 g/L with 78% fermentation efficiency was achieved from modified simultaneous saccharification and fermentation using cofermentation of the enzymatic hydrolysate of 300 g raw cassava chips/L with cane molasses.

A kinetic model and simulation of starch saccharification and simultaneous ethanol fermentation by amyloglucosidase and Zymomonas mobilis

Energy Technology Data Exchange (ETDEWEB)

Lee, C G [Michigan Univ., Ann Arbor, MI (United States). Dept. of Chemical Engineering; Kim, C H; Rhee, S K [Korea Inst. of Science and Technology, Taejon (Korea, Republic of). Genetic Engineering Research Inst.

1992-07-01

A mathematical model is described for the simultaneous saccharification and ethanol fermentation (SSF) of sago starch using amyloglycosidase (AMG) and Zymomonas mobilis. By introducing the degree of polymerization (DP) of oligosaccharides produced from sago starch treated with {alpha}-amylase, a series of Michaelis-Menten equations was obtained. After determining kinetic parameters from the results of simple experiments and from the subsite mapping theory, this model was adapted to simulate the SSF process. The results of simulation for SSF are in good agreement with experimental results. (orig.).

Simultaneous saccharification and ethanol fermentation at high corn stover solids loading in a helical stirring bioreactor.

Science.gov (United States)

Zhang, Jian; Chu, Deqiang; Huang, Juan; Yu, Zhanchun; Dai, Gance; Bao, Jie

2010-03-01

The higher ethanol titer inevitably requires higher solids loading during the simultaneous enzymatic saccharification and fermentation (SSF) using lignocellulose as the feedstock. The mixing between the solid lignocellulose and the liquid enzyme is crucially important. In this study, a bioreactor with a novel helical impeller was designed and applied to the SSF operation of the steam explosion pretreated corn stover under different solids loadings and different enzyme dosages. The performances using the helical impeller and the common Rushton impeller were compared and analyzed by measuring rheological properties and the mixing energy consumption. The results showed that the new designed stirring system had better performances in the saccharification yield, ethanol titer, and energy cost than those of the Rushton impeller stirring. The mixing energy consumption under different solids loadings and enzyme dosages during SSF operation were analyzed and compared to the thermal energy in the ethanol produced. A balance for achieving the optimal energy cost between the increased mixing energy cost and the reduced distillation energy cost at the high solids loading should be made. The potentials of the new bioreactor were tested under various SSF conditions for obtaining optimal ethanol yield and titer. (c) 2009 Wiley Periodicals, Inc.

Simultaneous saccharification of inulin and starch using commercial glucoamylase and the subsequent bioconversion to high titer sorbitol and gluconic acid.

Science.gov (United States)

An, Kehong; Hu, Fengxian; Bao, Jie

2013-12-01

A new bioprocess for production of sorbitol and gluconic acid from two low-cost feedstocks, inulin and cassava starch, using a commercially available enzyme was proposed in this study. The commercial glucoamylase GA-L NEW from Genencor was found to demonstrate a high inulinase activity for hydrolysis of inulin into fructose and glucose. The glucoamylase was used to replace the expensive and not commercially available inulinase enzyme for simultaneous saccharification of inulin and starch into high titer glucose and fructose hydrolysate. The glucose and fructose in the hydrolysate were converted into sorbitol and gluconic acid using immobilized whole cells of the recombinant Zymomonas mobilis strain. The high gluconic acid concentration of 193 g/L and sorbitol concentration of 180 g/L with the overall yield of 97.3 % were obtained in the batch operations. The present study provided a practical production method of sorbitol and gluconic acid from low cost feedstocks and enzymes.

Steam gasification of a thermally pretreated high lignin corn stover simultaneous saccharification and fermentation digester residue

Energy Technology Data Exchange (ETDEWEB)

Howe, Daniel T.; Taasevigen, Danny; Garcia-Perez, Manuel; McDonald, Armando G.; Li, Guosheng; Wolcott, Michael

2017-01-01

Efficient conversion of all components in lignocellulosic biomass is essential to realizing economic feasibility of biorefineries. However, when utilizing biochemical pathways, lignin cannot be fermented. Furthermore, the high lignin and high ash residue resulting from simultaneous saccharification and fermentation (SSF) reactors is difficult to thermochemically process due to feed line plugging and bed agglomeration. In this study a corn stover SSF digester residue was thermally pretreated at 300°C for 22.5 minutes (min) and then gasified in a bubbling fluidized bed gasifier to study the effect of thermal pretreatment on its processing behavior. Untreated, pelletized SSF residue was gasified at the same conditions to establish the baseline processing behavior. Results indicate that the thermal pretreatment process removes a substantial portion of the polar and non-polar extractives, with a resultant increase in the concentration of lignin, cellulose, and ash. Feed line plugging was not observed, although bed agglomeration was occurring at similar rates for both feedstocks, suggesting that overall ash content is the most important factor affecting bed agglomeration. Benzene, phenol, and polyaromatic hydrocarbons in the tar were present at higher concentrations in the treated material, with higher tar loading in the product gas. Total product gas generation is lower for the treated material, although the overall gas composition does not change.

Conversion of Aqueous Ammonia-Treated Corn Stover to Lactic Acid by Simultaneous Saccharification and Cofermentation

Science.gov (United States)

Zhu, Yongming; Lee, Y. Y.; Elander, Richard T.

Treatment of corn stover with aqueous ammonia removes most of the structural lignin, whereas retaining the majority of the carbohydrates in the solids. After treatment, both the cellulose and hemicellulose in corn stover become highly susceptible to enzymatic digestion. In this study, corn stover treated by aqueous ammonia was investigated as the substrate for lactic acid production by simultaneous saccharification and cofermentation (SSCF). A commercial cellulase (Spezyme-CP) and Lactobacillus pentosus American Type Culture Collection (ATCC) 8041 (Spanish Type Culture Collection [CECT]-4023) were used for hydrolysis and fermentation, respectively. In batch SSCF operation, the carbohydrates in the treated corn stover were converted to lactic acid with high yields, the maximum lactic acid yield reaching 92% of the stoichiometric maximum based on total fermentable carbohydrates (glucose, xylose, and arabinose). A small amount of acetic acid was also produced from pentoses through the phosphoketolase pathway. Among the major process variables for batch SSCF, enzyme loading and the amount of yeast extract were found to be the key factors affecting lactic acid production. Further tests on nutrients indicated that corn steep liquor could be substituted for yeast extract as a nitrogen source to achieve the same lactic acid yield. Fed-batch operation of the SSCF was beneficial in raising the concentration of lactic acid to a maximum value of 75.0 g/L.

Requirements of Saccharomyces Cerevisiae,Y 10 for Bioconversion of Lignocellulose Substrates to Ethanol under Simultaneous Saccharification and Fermentation Processes

International Nuclear Information System (INIS)

Rady, A.H.; Younis, N.A.; Sidkey, N.M.; Ouda, S.M.

2006-01-01

Ethanol production increased gradually with increasing the incubation period to a maximum value at 72 hrs for rice straw, bagasse and CHW (Cellulosic hospital wastes) under simultaneous saccharification and fermentation technique (SSF). bagasse was the best substrate for maximum production . maximum Values of ethanol were recorded when crude cellulses were 1.79, 0.597 and 1.19 (FPU /ml fermentation medium) for substrates respectively. the optimum inoculum number of yeast was (9x10 8 free cells / ml for rice straw, (1.2x10 9 cells/ml) of immobilized and free yeast for bagasse and CHW respectively. Maximum yield was recorded with ph 5 at 30 degree C for the three substrates. Fe SO 4 .7H 2 O(0.05%) increased ethanol production from pretreated bagasse and CHW .L-Iysine increased the productivity for both bagasse and CHW. molasses (9 g/l) achieved the highest productivity from treated rice straw, while thiamine B1 (100 and 200 ppm) for treated bagsse and CHW respectively. Gamma rays at doses 0.05 and 0.8 K.Gy increased ethanol yield 7.5 and 2 % for treated bagasse and CHW respectively. Highest values recorded at 300,200 and 100 rpm. for treated rice straw, bagasse and CHW, respectively

Expression of a mutated SPT15 gene in Saccharomyces cerevisiae enhances both cell growth and ethanol production in microaerobic batch, fed-batch, and simultaneous saccharification and fermentations.

Science.gov (United States)

Seong, Yeong-Je; Park, Haeseong; Yang, Jungwoo; Kim, Soo-Jung; Choi, Wonja; Kim, Kyoung Heon; Park, Yong-Cheol

2017-05-01

The SPT15 gene encodes a Saccharomyces cerevisiae TATA-binding protein, which is able to globally control the transcription levels of various metabolic and regulatory genes. In this study, a SPT15 gene mutant (S42N, S78R, S163P, and I212N) was expressed in S. cerevisiae BY4741 (BSPT15-M3), of which effects on fermentative yeast properties were evaluated in a series of culture types. By applying different nitrogen sources and air supply conditions in batch culture, organic nitrogen sources and microaerobic condition were decided to be more favorable for both cell growth and ethanol production of the BSPT15-M3 strain than the control S. cerevisiae BY4741 strain expressing the SPT15 gene (BSPT15wt). Microaerobic fed-batch cultures of BSPT15-M3 with glucose shock in the presence of high ethanol content resulted in a 9.5-13.4% higher glucose consumption rate and ethanol productivity than those for the BSPT15wt strain. In addition, BSPT15-M3 showed 4.5 and 3.9% increases in ethanol productivity from cassava hydrolysates and corn starch in simultaneous saccharification and fermentation processes, respectively. It was concluded that overexpression of the mutated SPT15 gene would be a potent strategy to develop robust S. cerevisiae strains with enhanced cell growth and ethanol production abilities.

Production of l(+)-lactic acid from acid pretreated sugarcane bagasse using Bacillus coagulans DSM2314 in a simultaneous saccharification and fermentation strategy.

Science.gov (United States)

van der Pol, Edwin C; Eggink, Gerrit; Weusthuis, Ruud A

2016-01-01

Sugars derived from lignocellulose-rich sugarcane bagasse can be used as feedstock for production of l(+)-lactic acid, a precursor for renewable bioplastics. In our research, acid-pretreated bagasse was hydrolysed with the enzyme cocktail GC220 and fermented by the moderate thermophilic bacterium Bacillus coagulans DSM2314. Saccharification and fermentation were performed simultaneously (SSF), adding acid-pretreated bagasse either in one batch or in two stages. SSF was performed at low enzyme dosages of 10.5-15.8 FPU/g DW bagasse. The first batch SSF resulted in an average productivity of 0.78 g/l/h, which is not sufficient to compete with lactic acid production processes using high-grade sugars. Addition of 1 g/l furfural to precultures can increase B. coagulans resistance towards by-products present in pretreated lignocellulose. Using furfural-containing precultures, productivity increased to 0.92 g/l/h, with a total lactic acid production of 91.7 g in a 1-l reactor containing 20% W/W DW bagasse. To increase sugar concentrations, bagasse was solubilized with a liquid fraction, obtained directly after acid pretreatment. Solubilizing the bagasse fibres with water increased the average productivity to 1.14 g/l/h, with a total lactic acid production of 84.2 g in a 1-l reactor. Addition of bagasse in two stages reduced viscosity during SSF, resulting in an average productivity in the first 23 h of 2.54 g/l/h, similar to productivities obtained in fermentations using high-grade sugars. Due to fast accumulation of lactic acid, enzyme activity was repressed during two-stage SSF, resulting in a decrease in productivity and a slightly lower total lactic acid production of 75.6 g. In this study, it is shown that an adequate production of lactic acid from lignocellulose was successfully accomplished by a two-stage SSF process, which combines acid-pretreated bagasse, B. coagulans precultivated in the presence of furfural as microorganism, and GC220 as enzyme

Systematic optimization of fed-batch simultaneous saccharification and fermentation at high-solid loading based on enzymatic hydrolysis and dynamic metabolic modeling of Saccharomyces cerevisiae.

Science.gov (United States)

Unrean, Pornkamol; Khajeeram, Sutamat; Laoteng, Kobkul

2016-03-01

An integrative simultaneous saccharification and fermentation (SSF) modeling is a useful guiding tool for rapid process optimization to meet the techno-economic requirement of industrial-scale lignocellulosic ethanol production. In this work, we have developed the SSF model composing of a metabolic network of a Saccharomyces cerevisiae cell associated with fermentation kinetics and enzyme hydrolysis model to quantitatively capture dynamic responses of yeast cell growth and fermentation during SSF. By using model-based design of feeding profiles for substrate and yeast cell in the fed-batch SSF process, an efficient ethanol production with high titer of up to 65 g/L and high yield of 85 % of theoretical yield was accomplished. The ethanol titer and productivity was increased by 47 and 41 %, correspondingly, in optimized fed-batch SSF as compared to batch process. The developed integrative SSF model is, therefore, considered as a promising approach for systematic design of economical and sustainable SSF bioprocessing of lignocellulose.

Rapid saccharification for production of cellulosic biofuels.

Science.gov (United States)

Lee, Dae-Seok; Wi, Seung Gon; Lee, Soo Jung; Lee, Yoon-Gyo; Kim, Yeong-Suk; Bae, Hyeun-Jong

2014-04-01

The economical production of biofuels is hindered by the recalcitrance of lignocellulose to processing, causing high consumption of processing enzymes and impeding hydrolysis of pretreated lignocellulosic biomass. We determined the major rate-limiting factor in the hydrolysis of popping pre-treated rice straw (PPRS) by examining cellulase adsorption to lignin and cellulose, amorphogenesis of PPRS, and re-hydrolysis. Based on the results, equivalence between enzyme loading and the open structural area of cellulose was required to significantly increase productive adsorption of cellulase and to accelerate enzymatic saccharification of PPRS. Amorphogenesis of PPRS by phosphoric acid treatment to expand open structural area of the cellulose fibers resulted in twofold higher cellulase adsorption and increased the yield of the first re-hydrolysis step from 13% to 46%. The total yield from PPRS was increased to 84% after 3h. These results provide evidence that cellulose structure is one of major effects on the enzymatic hydrolysis. Copyright © 2014 Elsevier Ltd. All rights reserved.

Lignosulfonate and elevated pH can enhance enzymatic saccharification of lignocelluloses

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Wang ZJ

2013-01-01

Full Text Available Abstract Background Nonspecific (nonproductive binding (adsorption of cellulase by lignin has been identified as a key barrier to reduce cellulase loading for economical sugar and biofuel production from lignocellulosic biomass. Sulfite Pretreatment to Overcome Recalcitrance of Lignocelluloses (SPORL is a relatively new process, but demonstrated robust performance for sugar and biofuel production from woody biomass especially softwoods in terms of yields and energy efficiencies. This study demonstrated the role of lignin sulfonation in enhancing enzymatic saccharification of lignocelluloses – lignosulfonate from SPORL can improve enzymatic hydrolysis of lignocelluloses, contrary to the conventional belief that lignin inhibits enzymatic hydrolysis due to nonspecific binding of cellulase. Results The study found that lignosulfonate from SPORL pretreatment and from a commercial source inhibits enzymatic hydrolysis of pure cellulosic substrates at low concentrations due to nonspecific binding of cellulase. Surprisingly, the reduction in enzymatic saccharification efficiency of a lignocellulosic substrate was fully recovered as the concentrations of these two lignosulfonates increased. We hypothesize that lignosulfonate serves as a surfactant to enhance enzymatic hydrolysis at higher concentrations and that this enhancement offsets its inhibitive effect from nonspecific binding of cellulase, when lignosulfonate is applied to lignocellulosic solid substrates. Lignosulfonate can block nonspecific binding of cellulase by bound lignin on the solid substrates, in the same manner as a nonionic surfactant, to significantly enhance enzymatic saccharification. This enhancement is linearly proportional to the amount of lignosulfonate applied which is very important to practical applications. For a SPORL-pretreated lodgepole pine solid, 90% cellulose saccharification was achieved at cellulase loading of 13 FPU/g glucan with the application of its

Simultaneously saccharification and fermentation approach as a tool for enhanced fossil fuels biodesulfurization.

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Paixão, Susana M; Arez, Bruno F; Roseiro, José C; Alves, Luís

2016-11-01

Biodesulfurization can be a complementary technology to the hydrodesulfurization, the commonly physical-chemical process used for sulfur removal from crude oil. The desulfurizing bacterium Gordonia alkanivorans strain 1B as a fructophilic microorganism requires fructose as C-source. In this context, the main goal of this work was the optimization of a simultaneous saccharification and fermentation (SSF) approach using the Zygosaccharomyces bailii strain Talf1 crude enzymes with invertase activity and sucrose as a cheaper fructose-rich commercial C-source (50% fructose) towards dibenzothiophene (DBT) desulfurization by strain 1B. The determination of optimal conditions, for both sucrose hydrolysis and DBT desulfurization was carried out through two sequential experimental uniform designs according to the Doehlert distribution for two factors: pH (5.5-7.5) and temperature (28-38 °C), with the enzyme load of 1.16 U/g/L; and enzyme load (0-4 U/g/L) and temperature (28-38 °C), with pH at 7.5. Based on 2-hydroxybiphenyl production, the analysis of the response surfaces obtained pointed out for pH 7.5, 32 °C and 1.8 U/g/L as optimal conditions. Further optimized SSF of sucrose during the DBT desulfurization process permitted to attain a 4-fold enhanced biodesulfurization. This study opens a new focus of research through the exploitation of sustainable low cost sucrose-rich feedstocks towards a more economical viable bioprocess scale-up. Copyright © 2016 Elsevier Ltd. All rights reserved.

The effect of nonenzymatic protein on lignocellulose enzymatic hydrolysis and simultaneous saccharification and fermentation.

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Wang, Hui; Kobayashi, Shinichi; Hiraide, Hatsue; Cui, Zongjun; Mochidzuki, Kazuhiro

2015-01-01

Nonenzymatic protein was added to cellulase hydrolysis and simultaneous saccharification and fermentation (SSF) of different biomass materials. Adding bovine serum albumin (BSA) and corn steep before cellulase enhanced enzyme activity in solution and increased cellulose and xylose conversion rates. The cellulose conversion rate of filter paper hydrolysis was increased by 32.5 % with BSA treatment. When BSA was added before cellulase, the remaining activity in the solution was higher than that in a control without BSA pretreatment. During SSF with pretreated rice straw as the substrate, adding 1.0 mg/mL BSA increased the ethanol yield by 13.6 % and final xylose yield by 42.6 %. The results indicated that lignin interaction is not the only mechanism responsible for the positive BSA effect. BSA had a stabilizing effect on cellulase and relieved cumulative sugar inhibition of enzymatic hydrolysis of biomass materials. Thus, nonenzymatic protein addition represents a promising strategy in the biorefining of lignocellulose materials.

Modeling cereal starch hydrolysis during simultaneous saccharification and lactic acid fermentation; case of a sorghum-based fermented beverage, gowé.

Science.gov (United States)

Mestres, Christian; Bettencourt, Munanga de J C; Loiseau, Gérard; Matignon, Brigitte; Grabulos, Joël; Achir, Nawel

2017-10-01

Gowé is an acidic beverage obtained after simultaneous saccharification and fermentation (SSF) of sorghum. A previous paper focused on modeling the growth of lactic acid bacteria during gowé processing. This paper focuses on modeling starch amylolysis to build an aggregated SSF model. The activity of α-amylase was modeled as a function of temperature and pH, and the hydrolysis rates of both native and soluble starch were modeled via a Michaelis-Menten equation taking into account the maltose and glucose inhibition constants. The robustness of the parameter estimators was ensured by step by step identification in sets of experiments conducted with different proportions of native and gelatinized starch by modifying the pre-cooking temperature. The aggregated model was validated on experimental data and showed that both the pre-cooking and fermentation parameters, particularly temperature, are significant levers for controlling not only acid and sugar contents but also the expected viscosity of the final product. This generic approach could be used as a tool to optimize the sanitary and sensory quality of fermentation of other starchy products. Copyright © 2017 Elsevier Ltd. All rights reserved.

Cost-effective simultaneous saccharification and fermentation of l-lactic acid from bagasse sulfite pulp by Bacillus coagulans CC17.

Science.gov (United States)

Zhou, Jie; Ouyang, Jia; Xu, Qianqian; Zheng, Zhaojuan

2016-12-01

The main barriers to cost-effective lactic acid production from lignocellulose are the high cost of enzymes and the ineffective utilization of the xylose within the hydrolysate. In the present study, the thermophilic Bacillus coagulans strain CC17 was used for the simultaneous saccharification and fermentation (SSF) of bagasse sulfite pulp (BSP) to produce l-lactic acid. Unexpectedly, SSF by CC17 required approximately 33.33% less fungal cellulase than did separate hydrolysis and fermentation (SHF). More interestingly, CC17 can co-ferment cellobiose and xylose without any exogenous β-glucosidase in SSF. Moreover, adding xylanase could increase the concentration of lactic acid produced via SSF. Up to 110g/L of l-lactic acid was obtained using fed-batch SSF, resulting in a lactic acid yield of 0.72g/g cellulose. These results suggest that SSF using CC17 has a remarkable advantage over SHF and that a potentially low-cost and highly-efficient fermentation process can be established using this protocol. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effect of steam explosion on waste copier paper alone and in a mixed lignocellulosic substrate on saccharification and fermentation

Science.gov (United States)

Elliston, Adam; Wilson, David R.; Wellner, Nikolaus; Collins, Samuel R.A.; Roberts, Ian N.; Waldron, Keith W.

2015-01-01

This study evaluated steam (SE) explosion on the saccharification and simultaneous saccharification and fermentation (SSF) of waste copier paper. SE resulted in a colouration, a reduction in fibre thickness and increased water absorption. Changes in chemical composition were evident at severities greater than 4.24 resulting in a loss of xylose and the production of breakdown products known to inhibit fermentation (particularly formic acid and acetic acid). SE did not improve final yields of glucose or ethanol, and at severities 4.53 and 4.83 reduced yields probably due to the effect of breakdown products and fermentation inhibitors. However, at moderate severities of 3.6 and 3.9 there was an increase in initial rates of hydrolysis which may provide a basis for reducing processing times. Co-steam explosion of waste copier paper and wheat straw attenuated the production of breakdown products, and may also provide a basis for improving SSF of lignocellulose. PMID:25846183

Bioconversion of paper sludge to biofuel by simultaneous saccharification and fermentation using a cellulase of paper sludge origin and thermotolerant Saccharomyces cerevisiae TJ14

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Harashima Satoshi

2011-09-01

Full Text Available Abstract Background Ethanol production from paper sludge (PS by simultaneous saccharification and fermentation (SSF is considered to be the most appropriate way to process PS, as it contains negligible lignin. In this study, SSF was conducted using a cellulase produced from PS by the hypercellulase producer, Acremonium cellulolyticus C-1 for PS saccharification, and a thermotolerant ethanol producer Saccharomyces cerevisiae TJ14 for ethanol production. Using cellulase of PS origin minimizes biofuel production costs, because the culture broth containing cellulase can be used directly. Results When 50 g PS organic material (PSOM/l was used in SSF, the ethanol yield based on PSOM was 23% (g ethanol/g PSOM and was two times higher than that obtained by a separate hydrolysis and fermentation process. Cellulase activity throughout SSF remained at around 60% of the initial activity. When 50 to 150 g PSOM/l was used in SSF, the ethanol yield was 21% to 23% (g ethanol/g PSOM at the 500 ml Erlenmeyer flask scale. Ethanol production and theoretical ethanol yield based on initial hexose was 40 g/l and 66.3% (g ethanol/g hexose at 80 h, respectively, when 161 g/l of PSOM, 15 filter paper units (FPU/g PSOM, and 20% inoculum were used for SSF, which was confirmed in the 2 l scale experiment. This indicates that PS is a good raw material for bioethanol production. Conclusions Ethanol concentration increased with increasing PSOM concentration. The ethanol yield was stable at PSOM concentrations of up to 150 g/l, but decreased at concentrations higher than 150 g/l because of mass transfer limitations. Based on a 2 l scale experiment, when 1,000 kg PS was used, 3,182 kFPU cellulase was produced from 134.7 kg PS. Produced cellulase was used for SSF with 865.3 kg PS and ethanol production was estimated to be 51.1 kg. Increasing the yeast inoculum or cellulase concentration did not significantly improve the ethanol yield or concentration.

Semicontinuous saccharification of starch in alcohol production

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Danilyak, N.I.; Kaminskil, R.S.; Shvedov, A.D.

1959-05-21

The saccharification is accomplished with an enzyme preparation of Aspergillus oryzae. In the first stage, the starch is treated at 57 to 59/sup 0/ with a fermenting solution containing 1% enzyme based on the starch content. The second step is carried out in the fermenting solution containing 2.5% enzyme.

Fourier transform infrared imaging and microscopy studies of Pinus radiata pulps regarding the simultaneous saccharification and fermentation process

Energy Technology Data Exchange (ETDEWEB)

Castillo, Rosario del P., E-mail: rosariocastillo@udec.cl [Faculty of Pharmacy, University of Concepcion, Concepcion (Chile); Biotechnology Center, University of Concepcion, Concepcion (Chile); Araya, Juan [Faculty of Pharmacy, University of Concepcion, Concepcion (Chile); Biotechnology Center, University of Concepcion, Concepcion (Chile); Troncoso, Eduardo [Consorcio Bioenercel S.A, University of Concepcion, Concepcion (Chile); Vinet, Silenne; Freer, Juanita [Biotechnology Center, University of Concepcion, Concepcion (Chile); Faculty of Chemical Sciences, University of Concepcion, Concepcion (Chile)

2015-03-25

The distribution and chemical patterns of lignocellulosic components at microscopic scale and their effect on the simultaneous saccharification and fermentation process (SSF) in the production of bioethanol from Pinus radiata pulps were analyzed by the application of diverse microscopical techniques, including scanning electronic microscopy (SEM), confocal laser scanning microscopy (CLSM) and attenuated total reflectance (ATR) – Fourier transform infrared microspectroscopy. This last technique was accompanied with multivariate methods, including principal component analysis (PCA) and multivariate curve resolution with alternating least squares (MCR-ALS) to evaluate the distribution patterns and to generate pure spectra of the lignocellulosic components of fibers. The results indicate that the information obtained by the techniques is complementary (ultrastructure, confocality and chemical characterization) and that the distribution of components affects the SSF yield, identifying lignin coalescence droplets as a characteristic factor to increase the SSF yield. Therefore, multivariate analysis of the infrared spectra enabled the in situ identification of the cellulose, lignin and lignin-carbohydrates arrangements. These techniques could be used to investigate the lignocellulosic components distribution and consequently their recalcitrance in many applications where minimal sample manipulation and microscale chemical information is required.

Fourier transform infrared imaging and microscopy studies of Pinus radiata pulps regarding the simultaneous saccharification and fermentation process

International Nuclear Information System (INIS)

Castillo, Rosario del P.; Araya, Juan; Troncoso, Eduardo; Vinet, Silenne; Freer, Juanita

2015-01-01

The distribution and chemical patterns of lignocellulosic components at microscopic scale and their effect on the simultaneous saccharification and fermentation process (SSF) in the production of bioethanol from Pinus radiata pulps were analyzed by the application of diverse microscopical techniques, including scanning electronic microscopy (SEM), confocal laser scanning microscopy (CLSM) and attenuated total reflectance (ATR) – Fourier transform infrared microspectroscopy. This last technique was accompanied with multivariate methods, including principal component analysis (PCA) and multivariate curve resolution with alternating least squares (MCR-ALS) to evaluate the distribution patterns and to generate pure spectra of the lignocellulosic components of fibers. The results indicate that the information obtained by the techniques is complementary (ultrastructure, confocality and chemical characterization) and that the distribution of components affects the SSF yield, identifying lignin coalescence droplets as a characteristic factor to increase the SSF yield. Therefore, multivariate analysis of the infrared spectra enabled the in situ identification of the cellulose, lignin and lignin-carbohydrates arrangements. These techniques could be used to investigate the lignocellulosic components distribution and consequently their recalcitrance in many applications where minimal sample manipulation and microscale chemical information is required

Simultaneous saccharification and co-fermentation of peracetic acid pretreated sugar cane bagasse

Energy Technology Data Exchange (ETDEWEB)

Teixeira, L.C. [Fundacao Centro Tecnologico de Minas Gerais, Belo Horizonte (Brazil); Linden, J.C.; Schroeder, H.A. [Colorado State University, Fort Collins, CO (United States)

1999-07-01

Previous work in our laboratory has demonstrated that peracetic acid improves the enzymatic digestibility of lignocellulosic materials. From the same studies, use of dilute alkali solutions as a pre-pretreatment prior to peracetic acid lignin oxidation increases sugar conversion yields in a synergistic, not additive, manner. Deacetylation of xylan is conducted easily by use of dilute alkali solutions at mild conditions. In this paper, the effectiveness of peracetic acid pretreatment of sugar cane bagasse combined with an alkaline pre-pretreatment, is evaluated through simultaneous saccharification and co-fermentation (SSCF) procedures. A practical 92% of theoretical ethanol yield using recombinant Zymomonas mobilis CP4/pZB5 is achieved using 6% NaOH/I5% peracetic acid pretreated substrate. No sugar accumulation is observed during SSCF; the recombinant microorganism exhibits greater glucose utilization rates than those of xylose. Acetate levels at the end of the co-fermentations are less than 0.2% (w/v). Based on demonstrated reduction of acetyl groups of the biomass, alkaline pre-pretreatments help to reduce peracetic acid requirements. The influence of deacetylation is more pronounced in combined pretreatments using lower peracetic acid loadings. Stereochemical impediments of the acetyl groups in hemicellulase on the activity of specific enzymes may be involved. (author)

Development of over-production strain of saccharification enzyme and biomass pretreatment by proton beam irradiation

International Nuclear Information System (INIS)

Kim, S. O.; Lee, J. Y.; Song, Y. S.; Shin, H. S.

2009-04-01

- The first year : Pre-treatment of biomass by proton beam irradiation and characterization of the pretreated biomass by IR and SEM - The second year : Strain development by proton beam irradiation for the production of cellulase and hemicellulase - The third year : Optimization of Saccharification process by cellulase and hemicellulase

Study on saccharification of cellulosic wastes with bench scale test plant, (5)

International Nuclear Information System (INIS)

Kasai, Noboru; Tamada, Masao; Kumakura, Minoru

1989-05-01

This report completed the results that were obtained on the studies of continuous saccharification of radiation pretreated chaff with a saccharification equipment unit of bench scale test plant for cellulosic wastes. The problem on the continuous saccharification in bench scale and its countermeasure were clarified. The glucose concentration obtained in the continuous saccharification was examined from the point of a scale up effect. It was found that there are not a scale up effect between flask scale (100 ml) and bench scale (50 l) and then the same concentration of glucose was obtained in both scales. It was clarified that the contamination of the process let decrease markedly the concentration of produced glucose solution and brings on a large trouble for the saccharification. The addition of 1 % ethyl acetate made it possible to prevent the contamination of the saccharification process in flask scale. However, in the case of continuous saccharification in bench scale, the addition of ethyl acetate in nitrogen gas atmosphere was necessary to prevent the contamination. It was found that the solution of 1.7 % glucose concentration was continuously produced in the continuous saccharification with the most longest period for 26 days. It was, also, suggested that the selection of a suitable retention time is necessary to attain a high glucose productivity in the continuous saccharification. (author)

Statistical optimization of recycled-paper enzymatic hydrolysis for simultaneous saccharification and fermentation via central composite design.

Science.gov (United States)

Liu, Qing; Cheng, Ke-ke; Zhang, Jian-an; Li, Jin-ping; Wang, Ge-hua

2010-01-01

A central composite design of the response surface methodology (RSM) was employed to study the effects of temperature, enzyme concentration, and stirring rate on recycled-paper enzymatic hydrolysis. Among the three variables, temperature and enzyme concentration significantly affected the conversion efficiency of substrate, whereas stirring rate was not effective. A quadratic polynomial equation was obtained for enzymatic hydrolysis by multiple regression analysis using RSM. The results of validation experiments were coincident with the predicted model. The optimum conditions for enzymatic hydrolysis were temperature, enzyme concentration, and stirring rate of 43.1 degrees C, 20 FPU g(-1) substrate, and 145 rpm, respectively. In the subsequent simultaneous saccharification and fermentation (SSF) experiment under the optimum conditions, the highest 28.7 g ethanol l(-1) was reached in the fed-batch SSF when 5% (w/v) substrate concentration was used initially, and another 5% added after 12 h fermentation. This ethanol output corresponded to 77.7% of the theoretical yield based on the glucose content in the raw material.

Linkage Mapping of Stem Saccharification Digestibility in Rice.

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Bohan Liu

Full Text Available Rice is the staple food of almost half of the world population, and in excess 90% of it is grown and consumed in Asia, but the disposal of rice straw poses a problem for farmers, who often burn it in the fields, causing health and environmental problems. However, with increased focus on the development of sustainable biofuel production, rice straw has been recognized as a potential feedstock for non-food derived biofuel production. Currently, the commercial realization of rice as a biofuel feedstock is constrained by the high cost of industrial saccharification processes needed to release sugar for fermentation. This study is focused on the alteration of lignin content, and cell wall chemotypes and structures, and their effects on the saccharification potential of rice lignocellulosic biomass. A recombinant inbred lines (RILs population derived from a cross between the lowland rice variety IR1552 and the upland rice variety Azucena with 271 molecular markers for quantitative trait SNP (QTS analyses was used. After association analysis of 271 markers for saccharification potential, 1 locus and 4 pairs of epistatic loci were found to contribute to the enzymatic digestibility phenotype, and an inverse relationship between reducing sugar and lignin content in these recombinant inbred lines was identified. As a result of QTS analyses, several cell-wall associated candidate genes are proposed that may be useful for marker-assisted breeding and may aid breeders to produce potential high saccharification rice varieties.

Empirical evaluation of inhibitory product, substrate, and enzyme effects during the enzymatic saccharification of lignocellulosic biomass.

Science.gov (United States)

Smith, Benjamin T; Knutsen, Jeffrey S; Davis, Robert H

2010-05-01

The cellulose hydrolysis kinetics during batch enzymatic saccharification are typified by a rapid initial rate that subsequently decays, resulting in incomplete conversion. Previous studies suggest that changes associated with the solution, substrate, or enzymes may be responsible. In this work, kinetic experiments were conducted to determine the relative magnitude of these effects. Pretreated corn stover (PCS) was used as a lignocellulosic substrate likely to be found in a commercial saccharification process, while Avicel and Kraft lignin were used to create model substrates. Glucose inhibition was observed by spiking the reaction slurry with glucose during initial-rate experiments. Increasing the glucose concentration from 7 to 48 g/L reduced the cellulose conversion rate by 94%. When product sugars were removed using ultrafiltration with a 10 kDa membrane, the glucose-based conversion increased by 9.5%. Reductions in substrate reactivity with conversion were compared directly by saccharifying PCS and Avicel substrates that had been pre-reacted to different conversions. Reaction of substrate with a pre-conversion of 40% resulted in about 40% reduction in the initial rate of saccharification, relative to fresh substrate with identical cellulose concentration. Overall, glucose inhibition and reduced substrate reactivity appear to be dominant factors, whereas minimal reductions of enzyme activity were observed.

Farm Deployable Microbial Bioreactor for Fuel Ethanol Production

Energy Technology Data Exchange (ETDEWEB)

Okeke, Benedict [Auburn Univ., Montgomery AL (United States)

2016-03-30

Research was conducted to develop a farm and field deployable microbial bioreactor for bioethanol production from biomass. Experiments were conducted to select the most efficient microorganisms for conversion of plant fiber to sugars for fermentation to ethanol. Mixtures of biomass and surface soil samples were collected from selected sites in Alabama black belt counties (Macon, Sumter, Choctaw, Dallas, Montgomery, Lowndes) and other areas within the state of Alabama. Experiments were conducted to determine the effects of culture parameters on key biomass saccharifying enzymes (cellulase, beta-glucosidase, xylanase and beta-xylosidase). A wide-scale sampling of locally-grown fruits in Central Alabama was embarked to isolate potential xylose fermenting microorganisms. Yeast isolates were evaluated for xylose fermentation. Selected microorganisms were characterized by DNA based methods. Factors affecting enzyme production and biomass saccharification were examined and optimized in the laboratory. Methods of biomass pretreatment were compared. Co-production of amylolytic enzymes with celluloytic-xylanolytic enzymes was evaluated; and co-saccharification of a combination of biomass, and starch-rich materials was examined. Simultaneous saccharification and fermentation with and without pre-saccharifcation was studied. Whole culture broth and filtered culture broth simultaneous saccahrifcation and fermentation were compared. A bioreactor system was designed and constructed to employ laboratory results for scale up of biomass saccharification.

Thermophilic fungi as new sources for production of cellulases and xylanases with potential use in sugarcane bagasse saccharification.

Science.gov (United States)

de Cassia Pereira, J; Paganini Marques, N; Rodrigues, A; Brito de Oliveira, T; Boscolo, M; da Silva, R; Gomes, E; Bocchini Martins, D A

2015-04-01

To obtain new cellulases and xylanases from thermophilic fungi; evaluate their potential for sugarcane bagasse saccharification. Thirty-two heat-tolerant fungi were isolated from the environment, identified (morphological/molecular tools) and the production of the enzymes was evaluated by solid state fermentation using lignocellulosic materials as substrates. Myceliophthora thermophila JCP 1-4 was the best producer of endoglucanase (357·51 U g(-1) ), β-glucosidase (45·42 U g(-1) ), xylanase (931·11 U g(-1) ) and avicelase (3·58 U g(-1) ). These enzymes were most active at 55-70°C and stable at 30-60°C. Using crude enzymatic extract from M. thermophila JCP 1-4 to saccharify sugarcane bagasse pretreated with microwaves and glycerol, glucose and xylose yields obtained were 15·6 and 35·13% (2·2 and 1·95 g l(-1) ), respectively. All isolated fungi have potential to produce the enzymes; M. thermophila JCP 1-4 enzymatic extract have potential to be better explored in saccharification experiments. Pretreatment improved enzymatic saccharification, as sugar yields were much higher than those obtained from in natura bagasse. Myceliophthora thermophila JCP 1-4 produces avicelase (not commonly found among fungi; important to hydrolyse crystalline cellulose) and a β-glucosidase resistant to glucose inhibition, interesting characteristics for saccharification experiments. © 2015 The Society for Applied Microbiology.

On-site cellulase production and efficient saccharification of corn stover employing cbh2 overexpressing Trichoderma reesei with novel induction system.

Science.gov (United States)

Li, Yonghao; Zhang, Xiaoyue; Xiong, Liang; Mehmood, Muhammad Aamer; Zhao, Xinqing; Bai, Fengwu

2017-08-01

Although on-site cellulase production offers cost-effective saccharification of lignocellulosic biomass, low enzyme titer is still a barrier for achieving robustness. In the present study, a strain of T. reesei was developed for enhanced production of cellulase via overexpression of Cellobiohydrolase II. Furthermore, optimum enzyme production was achieved using a novel inducer mixture containing synthesized glucose-sophorose (MGD) and alkali pre-treated corn stover (APCS). Within 60h, a remarkably higher cellulase productivity and activity were achieved in the fed-batch fermentation using the optimized ratio of MGD and APCS in the inducer mixture, compared to those reported using cellulosic biomass as the sole inducer. After the enzyme production, APCS was added directly into the fermentation broth at 20% solid loading, which produced 122.5g/L glucose and 40.21g/L xylose, leading to the highest yield reported so far. The improved enzyme titers during on-site cellulase production would benefit cost-competitive saccharification of lignocellulosic biomass. Copyright © 2017 Elsevier Ltd. All rights reserved.

Enzymatic saccharification of brown seaweed for production of fermentable sugars.

Science.gov (United States)

Sharma, Sandeep; Horn, Svein Jarle

2016-08-01

This study shows that high drying temperatures negatively affect the enzymatic saccharification yield of the brown seaweed Saccharina latissima. The optimal drying temperature of the seaweed in terms of enzymatic sugar release was found to be 30°C. The enzymatic saccharification process was optimized by investigating factors such as kinetics of sugar release, enzyme dose, solid loading and different blend ratios of cellulases and an alginate lyase. It was found that the seaweed biomass could be efficiently hydrolysed to fermentable sugars using a commercial cellulase cocktail. The inclusion of a mono-component alginate lyase was shown to improve the performance of the enzyme blend, in particular at high solid loadings. At 25% dry matter loading a combined glucose and mannitol concentration of 74g/L was achieved. Copyright © 2016 Elsevier Ltd. All rights reserved.

A MATHEMATICAL PROGRAMMING APPROACH FOR THE REPRESENTATION OF SACCHARIFICATION AND FERMENTATION STAGES OF A BIOETHANOL PLANT

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Yailet Albernas-Carvajal

2015-10-01

Full Text Available The biorefineries concept from renewable sources has gained much attention in recent years because they improve sustainability with regard to fossil fuel refineries that are limited by the depletion of petroleum reserves. In this perspective, the production of ethanol from sugar cane bagasse is highly attractive because it reduces the fossil fuels consumption, the energy costs and the greenhouse gases emission. In this context, this paper aims to develop an optimal model design of an ethanol plant, considering bagasse pretreatment stages for subsequent simultaneous saccharification and fermentation (SSF. SSF variant, as its name suggests, has the advantage that enzymatic hydrolysis and fermentation stages are simultaneously carried out on the same equipment, obtaining directly the ethanol as a main product. The proposed approach is based on a mixed integer linear programming model which is optimized by GAMS-CPLEX package.

Influence of fiber degradation and concentration of fermentable sugars on simultaneous saccharification and fermentation of high-solids spruce slurry to ethanol.

Science.gov (United States)

Hoyer, Kerstin; Galbe, Mats; Zacchi, Guido

2013-10-08

Saccharification and fermentation of pretreated lignocellulosic materials, such as spruce, should be performed at high solids contents in order to reduce the cost of the produced bioethanol. However, this has been shown to result in reduced ethanol yields or a complete lack of ethanol production. Previous studies have shown inconsistent results when prehydrolysis is performed at a higher temperature prior to the simultaneous saccharification and fermentation (SSF) of steam-pretreated lignocellulosic materials. In some cases, a significant increase in overall ethanol yield was reported, while in others, a slight decrease in ethanol yield was observed. In order to investigate the influence of prehydrolysis on high-solids SSF of steam-pretreated spruce slurry, in the present study, the presence of fibers and inhibitors, degree of fiber degradation and initial fermentable sugar concentration has been studied. SSF of whole steam-pretreated spruce slurry at a solids content of 13.7% water-insoluble solids (WIS) resulted in a very low overall ethanol yield, mostly due to poor fermentation. The yeast was, however, able to ferment the washed slurry and the liquid fraction of the pretreated slurry. Performing prehydrolysis at 48°C for 22 hours prior to SSF of the whole pretreated slurry increased the overall ethanol yield from 3.9 to 62.1%. The initial concentration of fermentable sugars in SSF could not explain the increase in ethanol yield in SSF with prehydrolysis. Although the viscosity of the material did not appear to decrease significantly during prehydrolysis, the degradation of the fibers prior to the addition of the yeast had a positive effect on ethanol yield when using whole steam-pretreated spruce slurry. The results of the present study suggest that the increase in ethanol yield from SSF when performing prehydrolysis is a result of fiber degradation rather than a decrease in viscosity. The increased concentration of fermentable sugars at the beginning of the

Optimization of the Enzymatic Saccharification Process of Milled Orange Wastes

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Daniel Velasco

2017-08-01

Full Text Available Orange juice production generates a very high quantity of residues (Orange Peel Waste or OPW-50–60% of total weight that can be used for cattle feed as well as feedstock for the extraction or production of essential oils, pectin and nutraceutics and several monosaccharides by saccharification, inversion and enzyme-aided extraction. As in all solid wastes, simple pretreatments can enhance these processes. In this study, hydrothermal pretreatments and knife milling have been analyzed with enzyme saccharification at different dry solid contents as the selection test: simple knife milling seemed more appropriate, as no added pretreatment resulted in better final glucose yields. A Taguchi optimization study on dry solid to liquid content and the composition of the enzymatic cocktail was undertaken. The amounts of enzymatic preparations were set to reduce their impact on the economy of the process; however, as expected, the highest amounts resulted in the best yields to glucose and other monomers. Interestingly, the highest content in solid to liquid (11.5% on dry basis rendered the best yields. Additionally, in search for process economy with high yields, operational conditions were set: medium amounts of hemicellulases, polygalacturonases and β-glucosidases. Finally, a fractal kinetic modelling of results for all products from the saccharification process indicated very high activities resulting in the liberation of glucose, fructose and xylose, and very low activities to arabinose and galactose. High activity on pectin was also observed, but, for all monomers liberated initially at a fast rate, high hindrances appeared during the saccharification process.

Development of a commercial enzymes system for lignocellulosic biomass saccharification

Energy Technology Data Exchange (ETDEWEB)

Kumar, Manoj

2012-12-20

DSM Innovation Inc., in its four year effort was able to evaluate and develop its in-house DSM fungal cellulolytic enzymes system to reach enzyme efficiency mandates set by DoE Biomass program MYPP goals. DSM enzyme cocktail is uniquely active at high temperature and acidic pH, offering many benefits and product differentiation in 2G bioethanol production. Under this project, strain and process development, ratio optimization of enzymes, protein and genetic engineering has led to multitudes of improvement in productivity and efficiency making development of a commercial enzyme system for lignocellulosic biomass saccharification viable. DSM is continuing further improvement by additional biodiversity screening, protein engineering and overexpression of enzymes to continue to further lower the cost of enzymes for saccharification of biomass.

Sustaining fermentation in high-gravity ethanol production by feeding yeast to a temperature-profiled multifeed simultaneous saccharification and co-fermentation of wheat straw.

Science.gov (United States)

Westman, Johan O; Wang, Ruifei; Novy, Vera; Franzén, Carl Johan

2017-01-01

Considerable progress is being made in ethanol production from lignocellulosic feedstocks by fermentation, but negative effects of inhibitors on fermenting microorganisms are still challenging. Feeding preadapted cells has shown positive effects by sustaining fermentation in high-gravity simultaneous saccharification and co-fermentation (SSCF). Loss of cell viability has been reported in several SSCF studies on different substrates and seems to be the main reason for the declining ethanol production toward the end of the process. Here, we investigate how the combination of yeast preadaptation and feeding, cell flocculation, and temperature reduction improves the cell viability in SSCF of steam pretreated wheat straw. More than 50% cell viability was lost during the first 24 h of high-gravity SSCF. No beneficial effects of adding selected nutrients were observed in shake flask SSCF. Ethanol concentrations greater than 50 g L -1 led to significant loss of viability and prevented further fermentation in SSCF. The benefits of feeding preadapted yeast cells were marginal at later stages of SSCF. Yeast flocculation did not improve the viability but simplified cell harvest and improved the feasibility of the cell feeding strategy in demo scale. Cultivation at 30 °C instead of 35 °C increased cell survival significantly on solid media containing ethanol and inhibitors. Similarly, in multifeed SSCF, cells maintained the viability and fermentation capacity when the temperature was reduced from 35 to 30 °C during the process, but hydrolysis yields were compromised. By combining the yeast feeding and temperature change, an ethanol concentration of 65 g L -1 , equivalent to 70% of the theoretical yield, was obtained in multifeed SSCF on pretreated wheat straw. In demo scale, the process with flocculating yeast and temperature profile resulted in 5% (w/w) ethanol, equivalent to 53% of the theoretical yield. Multifeed SSCF was further developed by means of a

Wet oxidation treatment of organic household waste enriched with wheat straw for simultaneous saccharification and fermentation into ethanol

DEFF Research Database (Denmark)

Lissens, G.; Klinke, H.B.; Verstraete, W.

2004-01-01

Organic municipal solid waste enriched with wheat straw was subjected to wet-oxidation as a pre-treatment for subsequent enzymatic conversion and fermentation into bio-ethanol. The effect of tempera (185-195degrees C), oxygen pressure (3-12) and sodium carbonate (0-2 g l(-1)) addition on enzymatic...... in the treated waste could be converted into respectively hexose and pentose sugars compared to 46% for cellulose and 36% for hemicellulose in the raw waste. For all wet oxidation conditions tested, total carbohydrate recoveries were high (> 89%) and 44-66% of the original lignin could be converted into non......-toxic carboxylic acids mainly (2.2-4.5 % on DS basis). Simultaneous saccharification and fermentation (SSF) of the treated waste at 10% DS by Saccharomyces cerevisae yielded average ethanol concentrations of 16.5 to 22 g l(-1) for enzyme loadings of 5 and 25 FPU g(-1) DS, respectively. The cellulose to ethanol...

Mild chemical pretreatments are sufficient for complete saccharification of steam-exploded residues and high ethanol production in desirable wheat accessions.

Science.gov (United States)

Zahoor; Tu, Yuanyuan; Wang, Lingqiang; Xia, Tao; Sun, Dan; Zhou, Shiguang; Wang, Yanting; Li, Ying; Zhang, Heping; Zhang, Tong; Madadi, Meysam; Peng, Liangcai

2017-11-01

In this study, a combined pretreatment was performed in four wheat accessions using steam explosion followed with different concentrations of H 2 SO 4 or NaOH, leading to increased hexoses yields by 3-6 folds from enzymatic hydrolysis. Further co-supplied with 1% Tween-80, Talq90 and Talq16 accessions exhibited an almost complete enzymatic saccharification of steam-exploded (SE) residues after 0.5% H 2 SO 4 or 1% NaOH pretreatment, with the highest bioethanol yields at 18.5%-19.4%, compared with previous reports about wheat bioethanol yields at 11%-17% obtained under relatively strong pretreatment conditions. Furthermore, chemical analysis indicated that much enhanced saccharification in Talq90 and Talq16 may be partially due to their relatively low cellulose CrI and DP values and high hemicellulose Ara and H-monomer levels in raw materials and SE residues. Hence, this study has not only demonstrated a mild pretreatment technology for a complete saccharification, but it has also obtained the high ethanol production in desirable wheat accessions. Copyright © 2017 Elsevier Ltd. All rights reserved.

Improving simultaneous saccharification and co-fermentation of pretreated wheat straw using both enzyme and substrate feeding

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Palmqvist Benny

2010-08-01

Full Text Available Abstract Background Simultaneous saccharification and co-fermentation (SSCF has been recognized as a feasible option for ethanol production from xylose-rich lignocellulosic materials. To reach high ethanol concentration in the broth, a high content of water-insoluble solids (WIS is needed, which creates mixing problems and, furthermore, may decrease xylose uptake. Feeding of substrate has already been proven to give a higher xylose conversion than a batch SSCF. In the current work, enzyme feeding, in addition to substrate feeding, was investigated as a means of enabling a higher WIS content with a high xylose conversion in SSCF of a xylose-rich material. A recombinant xylose-fermenting strain of Saccharomyces cerevisiae (TMB3400 was used for this purpose in fed-batch SSCF experiments of steam-pretreated wheat straw. Results By using both enzyme and substrate feeding, the xylose conversion in SSCF could be increased from 40% to 50% in comparison to substrate feeding only. In addition, by this design of the feeding strategy, it was possible to process a WIS content corresponding to 11% in SSCF and obtain an ethanol yield on fermentable sugars of 0.35 g g-1. Conclusion A combination of enzyme and substrate feeding was shown to enhance xylose uptake by yeast and increase overall ethanol yield in SSCF. This is conceptually important for the design of novel SSCF processes aiming at high-ethanol titers. Substrate feeding prevents viscosity from becoming too high and thereby allows a higher total amount of WIS to be added in the process. The enzyme feeding, furthermore, enables keeping the glucose concentration low, which kinetically favors xylose uptake and results in a higher xylose conversion.

Development of over-production strain of saccharification enzyme and biomass pretreatment by proton beam irradiation

International Nuclear Information System (INIS)

Kim, S. W.; Lee, J. Y.; Song, Y. S.; Lee, S. J.; Shin, H. Y.; Kim, S. B.

2010-04-01

When lignocellulosic biomass converts to ethanol, enzyme takes lots of part of whole cost. Therefore, cellulase production is one of the important processes for the successful enzymatic conversion of cellulosic biomass to ethanol. Among cellulolytic enzymes, cellulase is multi-complex enzyme containing endo-glucanase, exo-glucanase and β-glucosidase. Cellulolyticfungi, Trichodema reesei is well known to produce the highest yields of cellulase. Especially, suitable cellulase composition was important for the effective saccharification of lignocellulosic biomass and strain having high level production of cellulase should be developed for hydrolysis. For efficient ethanol production, hemicellullase of Aspergillus also develop to use xylose generated from saccharification of biomass. In this study, pretreatment process of rice straw using proton beam irradiation (PBI) was carried out for enhancement of enzyme digestibility at different proton beam doses. Also, PBI pretreatment on ammonia soaking treated (SAA, Soaking aqueous ammonia) rice straw was conducted to solve the problem that is micro-structural inhibition of rice straw. Optimal dosages of proton beam on rice straw and SAA treated rice straw for efficient recovery of sugar were 15 KGy and 3 KGy, respectively. Enzymatic saccharification of PBI treated rice straw and SAA rice straw was conducted for the guidance of NREL standard procedure. Analysis using X-ray diffractometry (XRD) for crystallinity index was carried out and CrI found to be 33.38% of control and 35.72% of 15 KGy. Also, CrI was determined to be 67.11% of control and approximately 65.58% of 3 kGy dose in PBI pretreatment on SAA treated rice straw. The result of sugar recovery of both was approximately 70 % and 91 % of theoretical glucose contents, respectively. The initial reaction rate was increased from 7.610 -4 g·l -1 ·s -1 of 15 KGy (PBI pretreated rice straw) to 9.710 -4 g·l -1 ·s -1 (3 KGy PBI pretreated SAA rice straw). The selection of

Effect of surfactants on separate hydrolysis fermentation and simultaneous saccharification fermentation of pretreated lodgepole pine.

Science.gov (United States)

Tu, Maobing; Zhang, Xiao; Paice, Mike; McFarlane, Paul; Saddler, Jack N

2009-01-01

The effects of surfactants addition on enzymatic hydrolysis and subsequent fermentation of steam exploded lodgepole pine (SELP) and ethanol pretreated lodgepole pine (EPLP) were investigated in this study. Supplementing Tween 80 during cellulase hydrolysis of SELP resulted in a 32% increase in the cellulose-to-glucose yield. However, little improvement was obtained from hydrolyzing EPLP in the presence of the same amount of surfactant. The positive effect of surfactants on SELP hydrolysis led to an increase in final ethanol yield after the fermentation. It was found that the addition of surfactant led to a substantial increase in the amount of free enzymes in the 48 h hydrolysates derived from both substrates. The effect of surfactant addition on final ethanol yield of simultaneous saccharification and fermentation (SSF) was also investigated by using SELP in the presence of additional furfural and hydroxymethylfurfural (HMF). The results showed that the surfactants slightly increased the conversion rates of furfural and HMF during SSF process by Saccharomyces cerevisiae. The presence of furfural and HMF at the experimental concentrations did not affect the final ethanol concentration either. The strategy of applying surfactants in cellulase recycling to reduce enzyme cost is presented. (c) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009.

Comparison of separate hydrolysis and fermentation and simultaneous saccharification and fermentation processes for ethanol production from wheat straw by recombinant Escherichia coli strain FBR5

Energy Technology Data Exchange (ETDEWEB)

Saha, Badal C.; Nichols, Nancy N.; Qureshi, Nasib; Cotta, Michael A. [U.S. Department of Agriculture, Agricultural Research Services Peoria, IL (United States). Bioenergy Reserach Unit

2011-11-15

Ethanol production by recombinant Escherichia coli strain FBR5 from dilute acid pretreated wheat straw (WS) by separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) was studied. The yield of total sugars from dilute acid (0.5% H2SO4) pretreated (160 C, 10 min) and enzymatically saccharified (pH 5.0, 45 C, 72 h) WS (86 g/l) was 50.0 {+-} 1.4 g/l. The hydrolyzate contained 1,184 {+-} 19 mg furfural and 161 {+-} 1 mg hydroxymethyl furfural per liter. The recombinant E. coli FBR5 could not grow at all at pH controlled at 4.5 to 6.5 in the non-abated wheat straw hydrolyzate (WSH) at 35 C. However, it produced 21.9 {+-} 0.3 g ethanol from non-abated WSH (total sugars, 44.1 {+-} 0.4 g/l) in 90 h including the lag time of 24 h at controlled pH 7.0 and 35 C. The bioabatement of WS was performed by growing Coniochaeta ligniaria NRRL 30616 in the liquid portion of the pretreated WS aerobically at pH 6.5 and 30 C for 15 h. The bacterium produced 21.6 {+-} 0.5 g ethanol per liter in 40 h from the bioabated enzymatically saccharified WSH (total sugars, 44.1 {+-} 0.4 g) at pH 6.0. It produced 24.9 {+-} 0.3 g ethanol in 96 h and 26.7 {+-} 0.0 g ethanol in 72 h per liter from bioabated WSH by batch SSF and fed-batch SSF, respectively. SSF offered a distinct advantage over SHF with respect to reducing total time required to produce ethanol from the bioabated WS. Also, fed-batch SSF performed better than the batch SSF with respect to shortening the time requirement and increase in ethanol yield. (orig.)

Conditions for saccharification and fermentation of manioc mash

Energy Technology Data Exchange (ETDEWEB)

Nguen, D.C.; Velikaya, E.I.

1972-01-01

Saccharification time of manioc starch, proportion of enzyme preparations added (obtained from Aspergillus oryzae and Aspergillus awamori) and the effect of a nitrogenous medium on fermentation were studied. Extension of saccharification time led to inactivation of the enzymes which adversely affected fermentation. In 5 minutes saccharification with Aspergillus oryzae of material pureed by boiling, the alcohol yield was 38.71/ton of starch higher than when a 60-minute saccharification period was used. Saccharification with a combination of the 2 moulds produced a higher alcohol yield than when only Aspergillus oryzae was used. A high glucoamylase content in the mash gave a high alcohol yield. Nitrogenous substances must be added to the manioc mash to provide for the nutrition of the yeast cells; carbamide proved most efficient.

Optimization of Xylanase Production through Response Surface Methodology by Fusarium sp. BVKT R2 Isolated from forest soil and its applications in saccharification

Directory of Open Access Journals (Sweden)

Ramanjaneyulu Golla

2016-09-01

Full Text Available AbstractXylanses are hydrolytic enzymes with wide applications in several industries like biofuels, paper and pulp, deinking, food and feed. The present study was aimed at hitting at high yield xylanase producing fungi from natural resources. Two highest xylanase producing fungal isolates - Q12 and L1were picked from collection of 450 fungal cultures for the utilization of xylan. These fungal isolates - Q12 and L1 were identified basing on ITS gene sequencing analysis as Fusarium sp. BVKT R2 (KT119615 and Fusarium strain BRR R6 (KT119619, respectively with construction of phylogenetic trees. Fusarium sp. BVKT R2 was further optimized for maximum xylanase production and the interaction effects between variables on production of xylanase were studied through response surface methodology. The optimal conditions for maximal production of xylanase were sorbitol 1.5%, yeast extract 1.5%, pH of 5.0, Temperature of 32.5ºC, and agitation of 175 rpm. Under optimal conditions, the yields of xylanase production by Fusarium sp. BVKT R2 was as high as 4560 U/ml in SmF. Incubation of different lignocellulosic biomasses with crude enzyme of Fusarium sp. BVKT R2 at 37°C for 72 h could achieve about 45% saccharification. The results suggest that Fusarium sp. BVKT R2 has potential applications in saccharification process of biomass.Key words: Fusarium sp., Optimization, Response Surface Methodology, Saccharification, Submerged fermentation, Xylanase

THE INFLUENCE OF FIRST WORT PART AND AFTERWORTS ON SACCHARIFICATION OF WORT

Directory of Open Access Journals (Sweden)

Miriam Líšková

2011-02-01

Full Text Available Normal 0 21 false false false SK X-NONE X-NONE MicrosoftInternetExplorer4 Wort is a basic product of mashing, which forms the first intermediate in beer production and constitute the base of its final value. For qualitative value wort has the greatest impact grist per brew, which is a description of materials, they bring to brew extract and determine its the volume and concentration. The main component grist per brew for light and dark beers is stored pale malt and possibly a smaller proportion of adjuncts. The aim of our work was to assess the qualitative parameters of malt in terms of content extract and its impact on the amount of produced the first wort part and afterwort and their qualitative values expressed in % saccharification and volumes. We measured 3 types of malts with the content of the extract 75.2%, 76.1%, 77.2% in the original sample, which determined mainly reached saccharification of first part wort and other afterwort parts one and two. In terms attained of saccharification it was necessary to use on sparge of spent grains at afterwort number two only the amount of water, which would be not affect the total saccharification of wort and its qualitative parameters.doi:10.5219/114 

A Mathematical Model for Simultaneous Saccharification and Co-fermentation (SSCF) of C6 and C5 Sugars

DEFF Research Database (Denmark)

Morales Rodriguez, Ricardo; Gernaey, Krist; Meyer, Anne S.

2011-01-01

saccharification and co-fermentation (SSCF) of C6 and C5 sugars. The model is constructed by combining existing mathematical models for enzymatic hydrolysis and co-fermentation. An inhibition of ethanol on cellulose conversion is introduced in order to increase the reliability. The mathematical model for the SSCF...... is verified by comparing the model predictions with experimental data obtained from the ethanol production based on kraft paper mill sludge. When fitting the model to the data, only the yield coefficients for glucose and xylose metabolism were fine-tuned, which were found to be 0.43 g·g−1 (ethanol....../glucose) and 0.35 g·g−1 (ethanol/xylose) respectively. These promising validation results encourage further model application to evaluate different process configurations for lignocellulosic bioethanol technology....

Bed Agglomeration During the Steam Gasification of a High Lignin Corn Stover Simultaneous Saccharification and Fermentation (SSF) Digester Residue

Energy Technology Data Exchange (ETDEWEB)

Howe, Daniel T.; Taasevigen, Danny J.; Gerber, Mark A.; Gray, Michel J.; Fernandez, Carlos A.; Saraf, Laxmikant; Garcia-Perez, Manuel; Wolcott, Michael P.

2015-11-13

This research investigates the bed agglomeration phenomena during the steam gasification of a high lignin residue produced from the simultaneous saccharification and fermentation (SSF) of corn stover in a bubbling fluidized bed. The studies were conducted at 895°C using alumina as bed material. Biomass was fed at 1.5 kg/hr, while steam was fed to give a velocity equal to 2.5 times the minimum fluidization velocity, with a steam/carbon ratio of 0.9. The pelletized feedstock was co-fed with a cooling nitrogen stream to mitigate feed line plugging issues. Tar production was high at 50.3 g/Nm3, and the fraction of C10+ compounds was greater than that seen in the gasification of traditional lignocellulosic feedstocks. Carbon closures over 94 % were achieved for all experiments. Bed agglomeration was found to be problematic, indicated by pressure drop increases observed below the bed and upstream of the feed line. Two size categories of solids were recovered from the reactor, +60 mesh and -60 mesh. After a 2.75-hour experiment, 61.7 wt % was recovered as -60 mesh particles and 38.2 wt% of the recovered reactor solids were +60 mesh. A sizeable percentage, 31.8 wt%, was +20 mesh. The -60 mesh particles were mainly formed by the initial bed material (Al2O3). Almost 50 wt. % of the + 20 mesh particles was found to be formed by organics. The unreacted carbon remaining in the reactor resulted in a low conversion rate to product gas. ICP-AES, SEM, SEM-EDS, and XRD confirmed that the large agglomerates (+ 20 mesh) were not encapsulated bed material but rather un-gasified feedstock pellets with sand particles attached to it.

Interlinked population balance and cybernetic models for the simultaneous saccharification and fermentation of natural polymers.

Science.gov (United States)

Ho, Yong Kuen; Doshi, Pankaj; Yeoh, Hak Koon; Ngoh, Gek Cheng

2015-10-01

Simultaneous Saccharification and Fermentation (SSF) is a process where microbes have to first excrete extracellular enzymes to break polymeric substrates such as starch or cellulose into edible nutrients, followed by in situ conversion of those nutrients into more valuable metabolites via fermentation. As such, SSF is very attractive as a one-pot synthesis method of biological products. However, due to the co-existence of multiple biochemical steps, modeling SSF faces two major challenges. The first is to capture the successive chain-end and/or random scission of the polymeric substrates over time, which determines the rate of generation of various fermentable substrates. The second is to incorporate the response of microbes, including their preferential substrate utilization, to such a complex broth. Each of the above-mentioned challenges has manifested itself in many related areas, and has been competently but separately attacked with two diametrically different tools, i.e., the Population Balance Modeling (PBM) and the Cybernetic Modeling (CM), respectively. To date, they have yet to be applied in unison on SSF resulting in a general inadequacy or haphazard approaches to examine the dynamics and interactions of depolymerization and fermentation. To overcome this unsatisfactory state of affairs, here, the general linkage between PBM and CM is established to model SSF. A notable feature is the flexible linkage, which allows the individual PBM and CM models to be independently modified to the desired levels of detail. A more general treatment of the secretion of extracellular enzyme is also proposed in the CM model. Through a case study on the growth of a recombinant Saccharomyces cerevisiae capable of excreting a chain-end scission enzyme (glucoamylase) on starch, the interlinked model calibrated using data from the literature (Nakamura et al., Biotechnol. Bioeng. 53:21-25, 1997), captured features not attainable by existing approaches. In particular, the effect

Saccharification of cellulosics by Microbispora bispora

Energy Technology Data Exchange (ETDEWEB)

Waldron, Jr, C R; Eveleigh, D E

1986-09-01

The saccharification efficiency of cellulase from the thermophilic actinomycete Microbispora bispora was evaluated using commercially available feedstocks. The enzyme preparation was effective against refuse derived cellulose with 30% being converted to glucose in a 24 hour period. Pretreatment of the refuse with cadoxen resulted in an increase in saccharification efficiency to 70%.

Impact of Pretreatment Technologies on Saccharification and Isopentenol Fermentation of Mixed Lignocellulosic Feedstocks

Energy Technology Data Exchange (ETDEWEB)

Shi, Jian; George, Kevin W.; Sun, Ning; He, Wei; Li, Chenlin; Stavila, Vitalie; Keasling, Jay D.; Simmons, Blake A.; Lee, Taek Soon; Singh, Seema

2015-02-28

In order to enable the large-scale production of biofuels or chemicals from lignocellulosic biomass, a consistent and affordable year-round supply of lignocellulosic feedstocks is essential. Feedstock blending and/or densification offers one promising solution to overcome current challenges on biomass supply, i.e., low energy and bulk densities and significant compositional variations. Therefore, it is imperative to develop conversion technologies that can process mixed pelleted biomass feedstocks with minimal negative impact in terms of overall performance of the relevant biorefinery unit operations: pretreatment, fermentable sugar production, and fuel titers. We processed the mixture of four feedstocks—corn stover, switchgrass, lodgepole pine, and eucalyptus (1:1:1:1 on dry weight basis)—in flour and pellet form using ionic liquid (IL) 1-ethyl-3-methylimidazolium acetate, dilute sulfuric acid (DA), and soaking in aqueous ammonia (SAA) pretreatments. Commercial enzyme mixtures, including cellulases and hemicellulases, were then applied to these pretreated feedstocks at low to moderate enzyme loadings to determine hydrolysis efficiency. Results show significant variations on the chemical composition, crystallinity, and enzymatic digestibility of the pretreated feedstocks across the different pretreatment technologies studied. The advanced biofuel isopentenol was produced during simultaneous saccharification and fermentation (SSF) of pretreated feedstocks using an engineered Escherichia coli strain. Results show that IL pretreatment liberates the most sugar during enzymatic saccharification, and in turn led to the highest isopentenol titer as compared to DA and SAA pretreatments. This study provides insights on developing biorefinery technologies that produce advanced biofuels based on mixed feedstock streams.

A Weibull statistics-based lignocellulose saccharification model and a built-in parameter accurately predict lignocellulose hydrolysis performance.

Science.gov (United States)

Wang, Mingyu; Han, Lijuan; Liu, Shasha; Zhao, Xuebing; Yang, Jinghua; Loh, Soh Kheang; Sun, Xiaomin; Zhang, Chenxi; Fang, Xu

2015-09-01

Renewable energy from lignocellulosic biomass has been deemed an alternative to depleting fossil fuels. In order to improve this technology, we aim to develop robust mathematical models for the enzymatic lignocellulose degradation process. By analyzing 96 groups of previously published and newly obtained lignocellulose saccharification results and fitting them to Weibull distribution, we discovered Weibull statistics can accurately predict lignocellulose saccharification data, regardless of the type of substrates, enzymes and saccharification conditions. A mathematical model for enzymatic lignocellulose degradation was subsequently constructed based on Weibull statistics. Further analysis of the mathematical structure of the model and experimental saccharification data showed the significance of the two parameters in this model. In particular, the λ value, defined the characteristic time, represents the overall performance of the saccharification system. This suggestion was further supported by statistical analysis of experimental saccharification data and analysis of the glucose production levels when λ and n values change. In conclusion, the constructed Weibull statistics-based model can accurately predict lignocellulose hydrolysis behavior and we can use the λ parameter to assess the overall performance of enzymatic lignocellulose degradation. Advantages and potential applications of the model and the λ value in saccharification performance assessment were discussed. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Utilization of radiation technique on the saccharification and fermentation of biomass

International Nuclear Information System (INIS)

Kaetsu, I.; Kumakura, M.; Fujimura, T.; Yoshii, F.; Kojima, T.; Tamada, M.

1981-01-01

The application of irradiation technique to the process of saccharification and subsequent fermentation of cellulosic wastes such as chaff and rice straw to obtain ethanol, was investigated. It was found that when waste raw materials were irradiated by γ-ray or electron beam, they became accessible to the subsequent enzymatic saccharification reaction. Irradiation of 10 7 to 10 8 Rad was enough for this effect. Some kind of additives reduced necessary dosage for this pretreatment. Cellulase, Trichoderma reesei which produce cellulase, and yeast were immobilized as biocatalysts for biomass conversion by radiation-induced polymerization of glass-forming monomer at low temperature. The immobilized cellulase showed almost the same activity of glucose production as the native cellulase. Continuous saccharification reaction was carried out by using the immobilized cellulase. The immobilized Trichoderma reesei and the immobilized yeast showed almost the same activity as the intact biocatalysts. It was concluded that the continuous saccharification and subsequent fermentation could be carried out effectively by using the immobilized biocatalysts. Spinach chloroplasts were immobilized by the same method as the first step for the conversion of water into hydrogen gas using solar energy. The immobilized chloroplasts kept the O 2 evolution activity in storage more than 30 days at 4 0 C. (author)

Utilization of radiation technique on the saccharification and fermentation of biomass

Energy Technology Data Exchange (ETDEWEB)

Kaetsu, I.; Kumakura, M.; Fujimura, T.; Yoshii, F.; Kojima, T.; Tamada, M. (Japan Atomic Energy Research Inst., Takasaki, Gunma. Takasaki Radiation Chemistry Research Establishment)

1981-01-01

The application of irradiation technique to the process of saccharification and subsequent fermentation of cellulosic wastes such as chaff and rice straw to obtain ethanol, was investigated. It was found that when waste raw materials were irradiated by ..gamma..-ray or electron beam, they became accessible to the subsequent enzymatic saccharification reaction. Irradiation of 10/sup 7/ to 10/sup 8/ Rad was enough for this effect. Some kind of additives reduced necessary dosage for this pretreatment. Cellulase, Trichoderma reesei which produce cellulase, and yeast were immobilized as biocatalysts for biomass conversion by radiation-induced polymerization of glass-forming monomer at low temperature. The immobilized cellulase showed almost the same activity of glucose production as the native cellulase. Continuous saccharification reaction was carried out by using the immobilized cellulase. The immobilized Trichoderma reesei and the immobilized yeast showed almost the same activity as the intact biocatalysts. It was concluded that the continuous saccharification and subsequent fermentation could be carried out effectively by using the immobilized biocatalysts. Spinach chloroplasts were immobilized by the same method as the first step for the conversion of water into hydrogen gas using solar energy. The immobilized chloroplasts kept the O/sub 2/ evolution activity in storage more than 30 days at 4/sup 0/C.

Utilization of radiation technique on the saccharification and fermentation of biomass

Science.gov (United States)

Kaetsu, I.; Kumakura, M.; Fujimura, T.; Yoshii, F.; Kojima, T.; Tamada, M.

The application of irradiation technique to the process of saccharification and subsequent fermentation of cellulosic wastes such as chaff and rice straw to obtain ethanol, was investigated. It was found that when waste raw materials were irradiated by ?-ray or electron beam, they became accessible to the subsequent enzymatic saccharification reaction. Irradiation of 10 7-10 8 Rad was enough for this effect. Some kind of additives reduced necessary dosage for this pretreatment. Cellulase, Trichoderma reesei which produce cellulase, and yeast were immobilized as biocatalysts for biomass conversion by radiation-induced polymerization of glass-forming monomer at low temperature. The immobilized cellulase showed almost same activity of glucose production as the native cellulase. Continuous saccharification reaction was carried out by using the immobilized cellulase. The immobilized Trichoderma reesei and the immobilized yeast showed almost same activity as the intact biocatalysts. It was concluded that the continuous saccharification and subsequent fermentation could be carried out effectively by using the immobilized biocatalysts. Spinach chloroplasts were immobilized by the same method as the first step for the conversion of water into hydrogen gas using solar energy. The immobilized chloroplasts kept the O 2 evolution activity in storage more than 30 days at 4°C. Thermostatility of chloroplasts was also improved greatly by the immobilization.

Downregulation of RWA genes in hybrid aspen affects xylan acetylation and wood saccharification.

Science.gov (United States)

Pawar, Prashant Mohan-Anupama; Ratke, Christine; Balasubramanian, Vimal K; Chong, Sun-Li; Gandla, Madhavi Latha; Adriasola, Mathilda; Sparrman, Tobias; Hedenström, Mattias; Szwaj, Klaudia; Derba-Maceluch, Marta; Gaertner, Cyril; Mouille, Gregory; Ezcurra, Ines; Tenkanen, Maija; Jönsson, Leif J; Mellerowicz, Ewa J

2017-06-01

High acetylation of angiosperm wood hinders its conversion to sugars by glycoside hydrolases, subsequent ethanol fermentation and (hence) its use for biofuel production. We studied the REDUCED WALL ACETYLATION (RWA) gene family of the hardwood model Populus to evaluate its potential for improving saccharification. The family has two clades, AB and CD, containing two genes each. All four genes are expressed in developing wood but only RWA-A and -B are activated by master switches of the secondary cell wall PtNST1 and PtMYB21. Histochemical analysis of promoter::GUS lines in hybrid aspen (Populus tremula × tremuloides) showed activation of RWA-A and -B promoters in the secondary wall formation zone, while RWA-C and -D promoter activity was diffuse. Ectopic downregulation of either clade reduced wood xylan and xyloglucan acetylation. Suppressing both clades simultaneously using the wood-specific promoter reduced wood acetylation by 25% and decreased acetylation at position 2 of Xylp in the dimethyl sulfoxide-extracted xylan. This did not affect plant growth but decreased xylose and increased glucose contents in the noncellulosic monosaccharide fraction, and increased glucose and xylose yields of wood enzymatic hydrolysis without pretreatment. Both RWA clades regulate wood xylan acetylation in aspen and are promising targets to improve wood saccharification. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

Saccharification of Sugarcane Bagasse by Enzymatic Treatment for bioethanol production

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Ahmed, F. M.

2012-06-01

Full Text Available Aims: The escalating demands for traditional fossil fuels with unsecured deliverance and issues of climate change compel the researchers to develop alternative fuels like bioethanol. This study examines the prospect of biofuel production from high carbohydrate containing lignocellulosic material, e.g. sugarcane bagasse through biological means. Methodology and Results: Cellulolytic enzymes were collected from the culture filtrate of thermotolerant Trichodermaviride grown on variously pre-treated sugarcane bagasse. CMCase and FPase enzyme activities were determined as a measure of suitable substrate pre-treatment and optimum condition for cellulolytic enzyme production. The highest CMCase and FPase activity was found to be 1.217 U/ml and 0.109 U/ml respectively under the production conditions of 200 rpm, pH 4.0 and 50 °C using steamed NaOH treated bagasse as substrate. SEM was carried out to compare and confirm the activity of cellulolytic enzymes on sugarcane bagasse. Saccharification of pre-treated bagasse was carried out with crude enzymes together using a two-factor experimental design. Under optimized conditions the pre-treated bagasse was saccharified up to 42.7 % in 24 h. The hydrolysate was concentrated by heating to suitable concentration and then used for fermentation by an indigenous isolate of Saccharomyces cerevisiae. With 50 and 80 % brix containing liquor the concentration of alcohol was 0.579 % and 1.15 % respectively. Conclusion, significance and impact of study: This is the first report in Bangladesh for the production of cellulosicethanol using local isolates. Though the rate of alcohol production was very low, a great impetus in this field can maximize the production thereby meet the demand for fuel in future.

Parameter Estimation for Simultaneous Saccharification and Fermentation of Food Waste Into Ethanol Using Matlab Simulink

Science.gov (United States)

Davis, Rebecca Anne

The increase in waste disposal and energy costs has provided an incentive to convert carbohydrate-rich food waste streams into fuel. For example, dining halls and restaurants discard foods that require tipping fees for removal. An effective use of food waste may be the enzymatic hydrolysis of the waste to simple sugars and fermentation of the sugars to ethanol. As these wastes have complex compositions which may change day-to-day, experiments were carried out to test fermentability of two different types of food waste at 27° C using Saccharomyces cerevisiae yeast (ATCC4124) and Genencor's STARGEN™ enzyme in batch simultaneous saccharification and fermentation (SSF) experiments. A mathematical model of SSF based on experimentally matched rate equations for enzyme hydrolysis and yeast fermentation was developed in Matlab Simulink®. Using Simulink® parameter estimation 1.1.3, parameters for hydrolysis and fermentation were estimated through modified Michaelis-Menten and Monod-type equations with the aim of predicting changes in the levels of ethanol and glycerol from different initial concentrations of glucose, fructose, maltose, and starch. The model predictions and experimental observations agree reasonably well for the two food waste streams and a third validation dataset. The approach of using Simulink® as a dynamic visual model for SSF represents a simple method which can be applied to a variety of biological pathways and may be very useful for systems approaches in metabolic engineering in the future.

Enzymatic saccharification of high pressure assist-alkali pretreated cotton stalk and structural characterization.

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Du, Shuang-kui; Su, Xia; Yang, Weihua; Wang, Yanqin; Kuang, Meng; Ma, Lei; Fang, Dan; Zhou, Dayun

2016-04-20

Cotton stalk is a potential biomass for bioethanol production, while the conversion of direct saccharification or biotransformation of cotton stalk is extremely low due to the recalcitrant nature of lignocellulose. To enhance the enzymatic conversion of cotton stalks, the enzymatic saccharification parameters of high pressure assist-alkali pretreatment (HPAP) cotton stalk were optimized in the present study. Results indicated that a maximum reducing sugar yield of 54.7g/100g dry biomass cellulose was achieved at a substrate concentration of 2%, 100rpm agitation, 0.6g/g enzyme loading, 40°C hydrolysis temperature, 50h saccharification time, and pH 5.0. Scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy were used to identify structural changes in native, pretreated biomass and hydrolyzed residues. Structural analysis revealed large part of amorphous cellulose and partial crystalline cellulose in the HPAP cotton stalk were hydrolyzed during enzymatic treatment. HPAP cotton stalk can be used as a potential feed stock for bioethanol production. Copyright © 2015 Elsevier Ltd. All rights reserved.

Bioethanol production from residual lignocellulosic materials: A review – Part 2

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CRISTIAN-TEODOR BURUIANA

2013-08-01

Full Text Available Lignocellulosic material (LCM can be employed as feedstock for biorefineries, a concept related to industries designed to process biomass for producing chemicals, fuels and/or electrical power. According to this philosophy, LCM can be fractionated and the resulting fractions employed for specific applications. Bioethanol production from cellulosic fraction of LCM involves: hydrolysis of polysaccharides and fermentation of the monomers into bioethanol. Enzymatic hydrolysis is catalyzed by cellulolytic enzymes and fermentation is carried out by bacteria, yeasts or fungi. The main objective of this article is to review different process integration technologies for bioethanol production from LCM. This paper include: separate hydrolysis and fermentation (SHF, simultaneous saccharification and fermentation (SSF, and simultaneous saccharification and co-fermentation (SSCF methods. Furthermore, the fermentation process and a comparative data of cellulases, hemicellulases and ethanol producing-microorganisms were presented.

Effects of different cellulases on the release of phenolic acids from rice straw during saccharification.

Science.gov (United States)

Xue, Yiyun; Wang, Xiahui; Chen, Xingxuan; Hu, Jiajun; Gao, Min-Tian; Li, Jixiang

2017-06-01

Effects of different cellulases on the release of phenolic acids from rice straw during saccharification were investigated in this study. All cellulases tested increased the contents of phenolic acids during saccharification. However, few free phenolic acids were detected, as they were present in conjugated form after saccharification when the cellulases from Trichoderma reesei, Trichoderma viride and Aspergillus niger were used. On the other hand, phenolic acids were present in free form when the Acremonium cellulolyticus cellulase was used. Assays of enzyme activity showed that, besides high cellulase activity, the A. cellulolyticus cellulase exhibited high feruloyl esterase (FAE) activity. A synergistic interaction between FAE and cellulase led to the increase in free phenolic acids, and thus an increase in antioxidative and antiradical activities of the phenolic acids. Moreover, a cost estimation demonstrated the feasibility of phenolic acids as value-added products to reduce the total production cost of ethanol. Copyright © 2017 Elsevier Ltd. All rights reserved.

Saccharification of rice straw by cellulase from a local Trichoderma harzianum SNRS3 for biobutanol production.

Science.gov (United States)

Rahnama, Nooshin; Foo, Hooi Ling; Abdul Rahman, Nor Aini; Ariff, Arbakariya; Md Shah, Umi Kalsom

2014-12-12

Rice straw has shown to be a promising agricultural by-product in the bioconversion of biomass to value-added products. Hydrolysis of cellulose, a main constituent of lignocellulosic biomass, is a requirement for fermentable sugar production and its subsequent bioconversion to biofuels such as biobutanol. The high cost of commercial enzymes is a major impediment to the industrial application of cellulases. Therefore, the use of local microbial enzymes has been suggested. Trichoderma harzianum strains are potential CMCase and β-glucosidase producers. However, few researches have been reported on cellulase production by T. harzianum and the subsequent use of the crude cellulase for cellulose enzymatic hydrolysis. For cellulose hydrolysis to be efficiently performed, the presence of the whole set of cellulase components including exoglucanase, endoglucanase, and β-glucosidase at a considerable concentration is required. Biomass recalcitrance is also a bottleneck in the bioconversion of agricultural residues to value-added products. An effective pretreatment could be of central significance in the bioconversion of biomass to biofuels. Rice straw pretreated using various concentrations of NaOH was subjected to enzymatic hydrolysis. The saccharification of rice straw pretreated with 2% (w/v) NaOH using crude cellulase from local T. harzianum SNRS3 resulted in the production of 29.87 g/L reducing sugar and a yield of 0.6 g/g substrate. The use of rice straw hydrolysate as carbon source for biobutanol fermentation by Clostridium acetobutylicum ATCC 824 resulted in an ABE yield, ABE productivity, and biobutanol yield of 0.27 g/g glucose, 0.04 g/L/h and 0.16 g/g glucose, respectively. As a potential β-glucosidase producer, T. harzianum SNRS3 used in this study was able to produce β-glucosidase at the activity of 173.71 U/g substrate. However, for cellulose hydrolysis to be efficient, Filter Paper Activity at a considerable concentration is also required to initiate the

Saccharification of sunflower stalks using lignocellulases from a fungal consortium comprising Pholiota adiposa and Armillaria gemina.

Science.gov (United States)

Ramachandran, Priyadharshini; Kim, Tae-Su; Dhiman, Saurabh Sudha; Li, Jinglin; Park, Ji-Hyun; Choi, Joon-Ho; Kim, Jae Young; Kim, Dongwook; Lee, Jung-Kul

2015-09-01

Lignocellulases from Armillaria gemina and Pholiota adiposa are efficient in hydrolyzing aspen and poplar biomass, respectively. In the present study, lignocellulosic enzymes obtained from a fungal consortium comprising P. adiposa and A. gemina were used for the saccharification of sunflower stalks. Sunflower stalks were thermochemically pretreated using 2 % NaOH at 50 °C for 24 h. The saccharification process parameters including substrate concentration, enzyme loading, pH, and temperature were optimized using response surface methodology to improve the saccharification yield. The highest enzymatic hydrolysis (84.3 %) was obtained using the following conditions: enzyme loading 10 FPU/g-substrate, substrate 5.5 %, temperature 50 °C, and pH 4.5. The hydrolysis yield obtained using the enzymes from the fungal consortium was equivalent to that obtained using a mixture of commercial enzymes Celluclast and Novozyme β-glucosidase. Addition of up to 500 ppm of heavy metal ions (As, Cu, Fe, Mn, Ni, Pb, and Zn) during saccharification did not significantly affect the saccharification yield. Thus, the biomass grown for phytoremediation of heavy metals can be used for the production of reducing sugars followed by ethanol fermentation.

Process for whole cell saccharification of lignocelluloses to sugars using a dual bioreactor system

Science.gov (United States)

Lu, Jue [Okemos, MI; Okeke, Benedict [Montgomery, AL

2012-03-27

The present invention describes a process for saccharification of lignocelluloses to sugars using whole microbial cells, which are enriched from cultures inoculated with paper mill waste water, wood processing waste and soil. A three-member bacterial consortium is selected as a potent microbial inocula and immobilized on inedible plant fibers for biomass saccharification. The present invention further relates the design of a dual bioreactor system, with various biocarriers for enzyme immobilization and repeated use. Sugars are continuously removed eliminating end-product inhibition and consumption by cell.

Citric waste saccharification under different chemical treatments

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Carlos Eduardo de Farias Silva

2015-10-01

Full Text Available Second generation ethanol from lignocellulose materials has been used in applications for food processing wastes. Since Brazil has a leading position in orange juice exports, the influence of acid and alkali pretreatments on liquor saccharification, solubilization of solid fraction and mass yield was evaluated. Time and Cacid or Calkaline at different concentrations of solids (low to moderate, 1 to 9% and high catalyst concentrations were analyzed. A hydrothermal pretreatment was conducted under the same conditions of acid and alkaline treatments to investigate the relative selectivity increase in using the catalysts. The chemical analyses of wastes indicated a 70% total carbohydrate level denoting a promising raw material for bioethanol production. Pretreatment caused acid saccharifications between 25 and 65% in total reducing sugars (TRS and mass yields (MY between 30 and 40%. In alkaline pretreatment, these rates ranged between 2 and 22.5% and between 30 and 80, respectively. In hydrothermal pretreatment, solubilized TRS varied between 3 and 37%, whereas MY remained between 45 and 60%, respectively. Cbiomass strongly influenced the three variables; in the same way, time affected MY.

Influence of acid phosphatase activity on the saccharification of potato maltodextrins by Aspergillus niger glucoamylase

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Zyla, K. (Akademia Rolnicza, Cracow (Poland). Dept. of Biotechnology)

1990-01-01

A preparation of Aspergillus niger acid phosphatase, which had the temperature optimum 60deg C, pH optimum 1.8-3.0; good stability at pH 4-5, the ability to hydrolyze glucose-6-phosphate at a high rate, and substantial lack of glucogenic activities, was used simultaneously with a glucoamylase in order to learn its influence on the saccharification of potato maltodextrins. The addition of the acid phosphatase activity in amounts that gave the 50 fold increase, as compared to phosphatase activity which naturally occurs in the gluocoamylase (GA) preparation 'AMG-200', was found to influence on the DE level, mainly at the high substrate concentration (40% d.s.) and low glucoamylase dosage (60-100 GAU/kg d.s.). It may also be possible, when using the acid phosphatase addition, to shorten the saccharification time. (orig.).

Substrate-Related Factors Affecting Enzymatic Saccharification of Lignocelluloses: Our Recent Understanding

Science.gov (United States)

Shao-Yuan Leu; J.Y. Zhu

2013-01-01

Enzymatic saccharification of cellulose is a key step in conversion of plant biomass to advanced biofuel and chemicals. Many substrate-related factors affect saccharification. Rather than examining the role of each individual factor on overall saccharification efficiency, this study examined how each factor affects the three basic processes of a heterogeneous...

Analysis, pretreatment and enzymatic saccharification of different fractions of Scots pine

Science.gov (United States)

2014-01-01

Background Forestry residues consisting of softwood are a major lignocellulosic resource for production of liquid biofuels. Scots pine, a commercially important forest tree, was fractionated into seven fractions of chips: juvenile heartwood, mature heartwood, juvenile sapwood, mature sapwood, bark, top parts, and knotwood. The different fractions were characterized analytically with regard to chemical composition and susceptibility to dilute-acid pretreatment and enzymatic saccharification. Results All fractions were characterized by a high glucan content (38-43%) and a high content of other carbohydrates (11-14% mannan, 2-4% galactan) that generate easily convertible hexose sugars, and by a low content of inorganic material (0.2-0.9% ash). The lignin content was relatively uniform (27-32%) and the syringyl-guaiacyl ratio of the different fractions were within the range 0.021-0.025. The knotwood had a high content of extractives (9%) compared to the other fractions. The effects of pretreatment and enzymatic saccharification were relatively similar, but without pretreatment the bark fraction was considerably more susceptible to enzymatic saccharification. Conclusions Since sawn timber is a main product from softwood species such as Scots pine, it is an important issue whether different parts of the tree are equally suitable for bioconversion processes. The investigation shows that bioconversion of Scots pine is facilitated by that most of the different fractions exhibit relatively similar properties with regard to chemical composition and susceptibility to techniques used for bioconversion of woody biomass. PMID:24641769

Methods of saccharification of polysaccharides in plants

Science.gov (United States)

Howard, John; Fake, Gina

2014-04-29

Saccharification of polysaccharides of plants is provided, where release of fermentable sugars from cellulose is obtained by adding plant tissue composition. Production of glucose is obtained without the need to add additional .beta.-glucosidase. Adding plant tissue composition to a process using a cellulose degrading composition to degrade cellulose results in an increase in the production of fermentable sugars compared to a process in which plant tissue composition is not added. Using plant tissue composition in a process using a cellulose degrading enzyme composition to degrade cellulose results in decrease in the amount of cellulose degrading enzyme composition or exogenously applied cellulase required to produce fermentable sugars.

Cassava as feedstock for ethanol production in South Africa | Marx ...

African Journals Online (AJOL)

It can be grown on marginal lands where frost is not prevalent. In this study, the production of ethanol from unpeeled Cassava roots and cassava peels were investigated. It was found that temperature; pH and biomass loading had a significant effect on glucose yield during hydrolysis. Simultaneous saccharification and ...

Efficient production of L-lactic acid by newly isolated thermophilic Bacillus coagulans WCP10-4 with high glucose tolerance.

Science.gov (United States)

Zhou, Xingding; Ye, Lidan; Wu, Jin Chuan

2013-05-01

A thermophilic Bacillus coagulans WCP10-4 with tolerance to high concentration of glucose was isolated from soil and used to produce optically pure L-lactic acid from glucose and starch. In batch fermentation at pH 6.0, 240 g/L of glucose was completely consumed giving 210 g/L of L-lactic acid with a yield of 95 % and a productivity of 3.5 g/L/h. In simultaneous saccharification and fermentation at 50 °C without sterilizing the medium, 200 g/L of corn starch was completely consumed producing 202.0 g/L of L-lactic acid. To the best of our knowledge, this strain shows the highest osmotic tolerance to glucose among the strains ever reported for lactic acid production. This is the first report of simultaneous saccharification and fermentation of starch for lactic acid production under a non-sterilized condition.

Do new cellulolytic enzyme preparations affect the industrial strategies for high solids lignocellulosic ethanol production?

DEFF Research Database (Denmark)

Cannella, David; Jørgensen, Henning

2014-01-01

proven essential for economic feasibility at industrial scale. Historically, simultaneous saccharification and fermentation (SSF) was found to give better ethanol yields compared to separate hydrolysis and fermentation (SHF), but data in literature are typically based on operating the process at low dry...... matter conditions. In this work the impact of selected enzyme preparation and processing strategy (SHF, presaccharification and simultaneous saccharification and fermentation—PSSF, and SSF) on final ethanol yield and overall performance was investigated with pretreated wheat straw up to 30% DM...... cellulose to around 94%, revealing that the most relevant products could be accounted for. One observation was the presence of oxidized sugar (gluconic acid) upon enzymatic hydrolysis with the latest enzyme preparation. Experiments showed gluconic acid formation by recently discovered enzymatic class...

Saccharification and fermentation of whole barley ground in the Szego mill

Energy Technology Data Exchange (ETDEWEB)

Wayman, M; Parekh, S R; Parekh, R S; Trass, O; Gandolfi, E

1988-11-01

Barley, after steeping in water, was ground with ease and efficiency in the Szego mill, and its starch was liquefied, saccharified and fermented to very high yields of ethanol. The Szego mill consists of vertical rollers with helical grooves which rotate within a fixed cylinder, resulting in very fine grinding and a somewhat flaky product. The steeped barley was ground to a fine paste. This was readily liquefied and saccharified by amylolytic enzymes (dual enzyme process), and the resulting sugars were fermented in 24 h by ordinary bakers' yeast Saccharomyces cerevisiae, resulting in over 450 l ethanol/t of barley. Still shorter time, 12 h, and the same high yield were achieved when liquefied barley starch was simultaneously saccharified by glucoamylase and fermented. Fermentation to ethanol by a glucoamylase-producing yeast S. diastaticus strain 164A (from Labatt Brewing Company) enabled the amount of this enzyme required for saccharification to be reduced to about one-half the normal quantity, but at some cost in slower fermentation and slightly lower ethanol yield.

Production of ethanol 3G from Kappaphycus alvarezii: evaluation of different process strategies.

Science.gov (United States)

Hargreaves, Paulo Iiboshi; Barcelos, Carolina Araújo; da Costa, Antonio Carlos Augusto; Pereira, Nei

2013-04-01

This study evaluated the potential of Kappaphycus alvarezii as feedstock for ethanol production, i.e. ethanol 3G. First, aquatic biomass was subjected to a diluted acid pretreatment. This acid pretreatment generated two streams--a galactose-containing liquid fraction and a cellulose-containing solid fraction, which were investigated to determine their fermentability with the following strategies: a single-stream process (simultaneous saccharification and co-fermentation (SSCF) of both fractions altogether), which achieved 64.3 g L(-1) of ethanol, and a two-stream process (fractions were fermented separately), which resulted in 38 g L(-1) of ethanol from the liquid fraction and 53.0 g L(-1) from the simultaneous saccharification and fermentation (SSF) of the solid fraction. Based on the average fermentable carbohydrate concentration, it was possible to obtain 105 L of ethanol per ton of dry seaweed. These preliminaries results indicate that the use of the macro-algae K. alvarezii has a good potential feedstock for bioethanol production. Copyright © 2013. Published by Elsevier Ltd.

Improvement of enzymatic saccharification yield in Arabidopsis thaliana by ectopic expression of the rice SUB1A-1 transcription factor

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Lizeth Núñez-López

2015-03-01

Full Text Available Saccharification of polysaccharides releases monosaccharides that can be used by ethanol-producing microorganisms in biofuel production. To improve plant biomass as a raw material for saccharification, factors controlling the accumulation and structure of carbohydrates must be identified. Rice SUB1A-1 is a transcription factor that represses the turnover of starch and postpones energy-consuming growth processes under submergence stress. Arabidopsis was employed to test if heterologous expression of SUB1A-1 or SUB1C-1 (a related gene can be used to improve saccharification. Cellulolytic and amylolytic enzymatic treatments confirmed that SUB1A-1 transgenics had better saccharification yield than wild-type (Col-0, mainly from accumulated starch. This improved saccharification yield was developmentally controlled; when compared to Col-0, young transgenic vegetative plants yielded 200–300% more glucose, adult vegetative plants yielded 40–90% more glucose and plants in reproductive stage had no difference in yield. We measured photosynthetic parameters, starch granule microstructure, and transcript abundance of genes involved in starch degradation (SEX4, GWD1, juvenile transition (SPL3-5 and meristematic identity (FUL, SOC1 but found no differences to Col-0, indicating that starch accumulation may be controlled by down-regulation of CONSTANS and FLOWERING LOCUS T by SUB1A-1 as previously reported. SUB1A-1 transgenics also offered less resistance to deformation than wild-type concomitant to up-regulation of AtEXP2 expansin and BGL2 glucan-1,3,-beta-glucosidase. We conclude that heterologous SUB1A-1 expression can improve saccharification yield and softness, two traits needed in bioethanol production.

Simultaneous saccharification and co-fermentation of paper sludge to ethanol by Saccharomyces cerevisiae RWB222--Part I: kinetic modeling and parameters.

Science.gov (United States)

Zhang, Jiayi; Shao, Xiongjun; Townsend, Oliver V; Lynd, Lee R

2009-12-01

A kinetic model was developed to predict batch simultaneous saccharification and co-fermentation (SSCF) of paper sludge by the xylose-utilizing yeast Saccharomyces cerevisiae RWB222 and the commercial cellulase preparation Spezyme CP. The model accounts for cellulose and xylan enzymatic hydrolysis and competitive uptake of glucose and xylose. Experimental results show that glucan and xylan enzymatic hydrolysis are highly correlated, and that the low concentrations of xylose encountered during SSCF do not have a significant inhibitory effect on enzymatic hydrolysis. Ethanol is found to not only inhibit the specific growth rate, but also to accelerate cell death. Glucose and xylose uptake rates were found to be competitively inhibitory, but this did not have a large impact during SSCF because the sugar concentrations are low. The model was used to evaluate which constants had the greatest impact on ethanol titer for a fixed substrate loading, enzyme loading, and fermentation time. The cellulose adsorption capacity and cellulose hydrolysis rate constants were found to have the greatest impact among enzymatic hydrolysis related constants, and ethanol yield and maximum ethanol tolerance had the greatest impact among fermentation related constants.

Valorisation of food and beverage waste via saccharification for sugars recovery.

Science.gov (United States)

Kwan, Tsz Him; Ong, Khai Lun; Haque, Md Ariful; Kwan, Wing Hei; Kulkarni, Sandeep; Lin, Carol Sze Ki

2018-05-01

Valorisation of mixed food and beverage (F&B) waste was studied for the recovery of sugars via saccharification. Glucoamylase and sucrase were employed to hydrolyse the starch and sucrose present in the mixed F&B waste because of the high cost-effectiveness for such recovery. The Michaelis-Menten kinetics model suggests that preservatives and additives in beverages did not inhibit glucoamylase and sucrase during saccharification. High levels of glucose (228.1 g L -1 ) and fructose (55.7 g L -1 ) were efficiently produced within 12 h at a solid-to-liquid ratio of 37.5% (w/v) in 2.5 L bioreactors. An overall conversion yield of 0.17 g sugars per g of mixed F&B waste was obtained in mass balance analysis. Lastly, possible industrial applications of the sugar-rich hydrolysate and by-products are discussed. This study is believed to cast insights into F&B waste recycling via biotechnology to produce high-value added products to promote the establishment of a circular bio-economy. Copyright © 2018 Elsevier Ltd. All rights reserved.

Kinetic modeling of cellulosic biomass to ethanol via simultaneous saccharification and fermentation: Part I. Accommodation of intermittent feeding and analysis of staged reactors.

Science.gov (United States)

Shao, Xiongjun; Lynd, Lee; Wyman, Charles; Bakker, André

2009-01-01

The model of South et al. [South et al. (1995) Enzyme Microb Technol 17(9): 797-803] for simultaneous saccharification of fermentation of cellulosic biomass is extended and modified to accommodate intermittent feeding of substrate and enzyme, cascade reactor configurations, and to be more computationally efficient. A dynamic enzyme adsorption model is found to be much more computationally efficient than the equilibrium model used previously, thus increasing the feasibility of incorporating the kinetic model in a computational fluid dynamic framework in the future. For continuous or discretely fed reactors, it is necessary to use particle conversion in conversion-dependent hydrolysis rate laws rather than reactor conversion. Whereas reactor conversion decreases due to both reaction and exit of particles from the reactor, particle conversion decreases due to reaction only. Using the modified models, it is predicted that cellulose conversion increases with decreasing feeding frequency (feedings per residence time, f). A computationally efficient strategy for modeling cascade reactors involving a modified rate constant is shown to give equivalent results relative to an exhaustive approach considering the distribution of particles in each successive fermenter.

Dried culture of the fungus Aspergillus oryzae for saccharification in alcohol production

Energy Technology Data Exchange (ETDEWEB)

Drazhner, T M; Ashkinuzi, Z K; El' chits, S V

1963-01-01

Dried A. oryzae in place of malt in saccharification of starch-containing grain mash is known to save valuable grain, labor, and space. Laboratory tests showed adequate amylolytic, dextrinolytic, and maltase activity even after nine months storage. Treatment with 0.1% formalin for one hour or with the filtrate of a calcium hypochlorite solution containing 700 mg/l active chlorine for two hours prevented growth of contaminating microflora for 24 hours of fermentation and was not detrimental. Reduction of dried culture needed for saccharification from 8 to 10 to 3.5 to 4% by weight of the starch in the raw material was possible by 2 methods: (1) a l-stage addition at 40/sup 0/, or (2) a 2-stage introduction: 20 to 25% into the sugaring vessel at 57 to 59/sup 0/ and the rest into the fermentation vat after yeast is added. Method 2 avoids the viscosity problem. Alcohol yield was essentially the same as with malt. An industrial test for a 66-hour fermentation confirmed the advantages of the 2-stage method. Higher acidity of final broth did not affect the results because mold enzymes are more resistant to this factor than are those of malt. Nor is higher dry content, the result of breakdown of hemicellulose, cellulose, and pentosans by mold enzymes, a marked disadvantage.

Bacterial cellulose production from cotton-based waste textiles: enzymatic saccharification enhanced by ionic liquid pretreatment.

Science.gov (United States)

Hong, Feng; Guo, Xiang; Zhang, Shuo; Han, Shi-fen; Yang, Guang; Jönsson, Leif J

2012-01-01

Cotton-based waste textiles were explored as alternative feedstock for production of bacterial cellulose (BC) by Gluconacetobacter xylinus. The cellulosic fabrics were treated with the ionic liquid (IL) 1-allyl-3-methylimidazolium chloride ([AMIM]Cl). [AMIM]Cl caused 25% inactivation of cellulase activity at a concentration as low as of 0.02 g/mL and decreased BC production during fermentation when present in concentrations higher than 0.0005 g/mL. Therefore, removal of residual IL by washing with hot water was highly beneficial to enzymatic saccharification as well as BC production. IL-treated fabrics exhibited a 5-7-fold higher enzymatic hydrolysis rate and gave a seven times larger yield of fermentable sugars than untreated fabrics. BC from cotton cloth hydrolysate was obtained at an yield of 10.8 g/L which was 83% higher than that from the culture grown on glucose-based medium. The BC from G. xylinus grown on IL-treated fabric hydrolysate had a 79% higher tensile strength than BC from glucose-based culture medium which suggests that waste cotton pretreated with [AMIM]Cl has potential to serve as a high-quality carbon source for BC production. Copyright © 2011 Elsevier Ltd. All rights reserved.

Ultrasonic pretreatment for enhanced saccharification and fermentation of ethanol production from corn

Science.gov (United States)

Montalbo-Lomboy, Melissa T.

during pretreatment. Ultrasonication of sugary-2 corn was also investigated in the study. Results similar to those for commodity corn (dent corn) were found, in terms of glucose yield and starch conversion. SEM and polarized-light microscope pictures showed the partial gelatinization of corn slurry due to ultrasound. In the 96-h saccharification time, a model was formulated to fit the sugar release curve. The results have shown 17-21% increase in the extent of sugar production from sonicated samples relative to the control group. Additionally, the reaction rates of the sonicated samples were 2- to 10-fold higher than the reaction rates for the control group. In comparing sugary-2 corn with commodity corn, it was found that sonicated sugary-2 corn saccharified faster than sonicated commodity corn. It is important to note, without ultrasonic treatment, sugary-2 corn released more reducing sugar than commodity corn during saccharification. To further investigate the potential of ultrasonics for scale-up, a continuous flow system was studied. An ultrasonic continuous flow system was tested using Branson's flow-through "donut" horn. The donut horn, which vibrates radially, was placed inside a 5.5 L stainless steel reactor. The amplitude was maintained at 12 mumpp and the feed flow rate was varied from 8-27 L/min (2-7 gal/min) with reactor retention times varying from 12-40 seconds. Samples sonicated in continuous flow system showed lower reducing sugar yield than batch ultrasonication. However, considering the ultrasonic energy density of batch and continuous systems, the continuous systems proved to be more energy efficient in terms of glucose production compared with the batch system. It was also seen that particle size disintegration was proportional to energy density regardless of the type of ultrasonic system used. To compare ultrasonics with jet cooking, fermentation experiments were conducted. There were only marginal differences between jet cooked samples and the

Enhanced bioprocessing of lignocellulose: Wood-rot fungal saccharification and fermentation of corn fiber to ethanol

Science.gov (United States)

Shrestha, Prachand

no improvement in ethanol yields. We showed that saccharification of lignocellulosic material with a wood-rot fungal process is quite feasible. Corn fiber from wet milling was best degraded to sugars using aerobic solid state fermentation with the soft-rot fungus T. reesei. However, it was shown that both the white-rot fungus P. chrysosporium and brown-rot fungus G. trabeum had the ability to produce additional consortia of hemi/cellulose degrading enzymes. It is likely that a consortium of enzymes from these fungi would be the best approach in saccharification of lignocellulose. In all cases, a subsequent anaerobic yeast process under submerged conditions is required to ferment the released sugars to ethanol. To our knowledge, this is the first time report on production of cellulolytic enzymes from wet-milled corn fiber using white- and brown-rot fungi for sequential fermentation of corn fiber hydrolyzate to ethanol. Keywords: lignocellulose, ethanol, biofuel, bioeconomy, biomass, renewable resources, corn fiber, pretreatment, solid-substrate fermentation, simultaneous saccharification and fermentation (SSF), white-rot fungus, brown-rot fungus, soft-rot fungus, fermentable sugars, enzyme activities, cellulytic enzymes Phanerochaete chrysosporium, Gloleophyllum trabeum, Trichoderma reesei, Saccharomyces cerevisiae.

Ethanol Production from Kitchen Garbage Using Zymomonas mobilis: Optimization of Parameters through Statistical Experimental Designs

OpenAIRE

Ma, H.; Wang, Q.; Gong, L.; Wang, X.; Yin, W.

2008-01-01

Plackett-Burman design was employed to screen 8 parameters for ethanol production from kitchen garbage by Zymomonas mobilis in simultaneous saccharification and fermentation. The parameters were divided into two parts, four kinds of enzymes and supplementation nutrients. The result indicated that the nutrient inside kitchen garbage could meet the requirement of ethanol production without supplementation, only protease and glucoamylase were needed to accelerate the ethanol production. The opti...

Ethanol and anaerobic conditions reversibly inhibit commercial cellulase activity in thermophilic simultaneous saccharification and fermentation (tSSF

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Podkaminer Kara K

2012-06-01

Full Text Available Abstract Background A previously developed mathematical model of low solids thermophilic simultaneous saccharification and fermentation (tSSF with Avicel was unable to predict performance at high solids using a commercial cellulase preparation (Spezyme CP and the high ethanol yield Thermoanaerobacterium saccharolyticum strain ALK2. The observed hydrolysis proceeded more slowly than predicted at solids concentrations greater than 50 g/L Avicel. Factors responsible for this inaccuracy were investigated in this study. Results Ethanol dramatically reduced cellulase activity in tSSF. At an Avicel concentration of 20 g/L, the addition of ethanol decreased conversion at 96 hours, from 75% in the absence of added ethanol down to 32% with the addition of 34 g/L initial ethanol. This decrease is much greater than expected based on hydrolysis inhibition results in the absence of a fermenting organism. The enhanced effects of ethanol were attributed to the reduced, anaerobic conditions of tSSF, which were shown to inhibit cellulase activity relative to hydrolysis under aerobic conditions. Cellulose hydrolysis in anaerobic conditions was roughly 30% slower than in the presence of air. However, this anaerobic inhibition was reversed by exposing the cellulase enzymes to air. Conclusion This work demonstrates a previously unrecognized incompatibility of enzymes secreted by an aerobic fungus with the fermentation conditions of an anaerobic bacterium and suggests that enzymes better suited to industrially relevant fermentation conditions would be valuable. The effects observed may be due to inactivation or starvation of oxygen dependent GH61 activity, and manipulation or replacement of this activity may provide an opportunity to improve biomass to fuel process efficiency.

EVALUATION OF BIOETHANOL PRODUCTION FROM Eucalyptus WOOD WITH Saccharomyces cerevisiae AND SACSV-10 1

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Sylvia Enid Vazquez

2018-04-01

Full Text Available ABSTRACT Eucalyptus spp. residues of paper industry are a potential lignocellulosic raw material for production of second-generation bioethanol as an alternative to conventional production from cereal crops. Studying the behavior at 40 ºC of a commercial cellulase (Sunson, Eucalyptus sawdust saccharification was carried out under two pH conditions. With the aim to evaluate the bioethanol production from Eucalyptus wood, a strategy combining saccharification and Simultaneous Saccharification and Fermentation (SSF was undertaken at 40 ºC with a thermotolerant Saccharomyces cerevisiae with different substrate and inoculum concentrations, and different nitrogen sources. At last, the process was carried out in optimal conditions with Saccharomyces cerevisiae M522 and SacSV-10. Saccharification produced more free glucose at pH 5, reaching a maximum of 1.5 g/L. Encouraging results were obtained with 500 mg/L of ammonium sulphate as a nitrogen source and 10 % v/v initial inoculum at 106 cfu/mL concentration. Yeast SacSV-10 was not inhibited by phenols present in the culture media using a wood concentration of 10 g/L, but when the solids concentration was increased, the bioprocess yield was compromised. When the process was carried out in optimal conditions the bioethanol production, expressed as the conversion percentage of cellulose to ethanol, was 71.5 % and 73.6 % for M522 and the mutant strain respectively. The studied properties of the mutant strain provide added value to it, which pose new challenges to national companies dedicated to the production and sale of inputs for bioethanol industry.

Saccharification Performances of Miscanthus at the Pilot and Miniaturized Assay Scales: Genotype and Year Variabilities According to the Biomass Composition

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Nassim Belmokhtar

2017-05-01

Full Text Available HIGHLIGHTSBiomass production and cell wall composition are differentially impacted by harvesting year and genotypes, influencing then cellulose conversion in miniaturized assay.Using a high-throughput miniaturized and semi-automated method for performing the pretreatment and saccharification steps at laboratory scale allows for the assessment of these factors on the biomass potential for producing bioethanol before moving to the industrial scale.The large genetic diversity of the perennial grass miscanthus makes it suitable for producing cellulosic ethanol in biorefineries. The saccharification potential and year variability of five genotypes belonging to Miscanthus × giganteus and Miscanthus sinensis were explored using a miniaturized and semi-automated method, allowing the application of a hot water treatment followed by an enzymatic hydrolysis. The studied genotypes highlighted distinct cellulose conversion yields due to their distinct cell wall compositions. An inter-year comparison revealed significant variations in the biomass productivity and cell wall compositions. Compared to the recalcitrant genotypes, more digestible genotypes contained higher amounts of hemicellulosic carbohydrates and lower amounts of cellulose and lignin. In contrast to hemicellulosic carbohydrates, the relationships analysis between the biomass traits and cellulose conversion clearly showed the same negative effect of cellulose and lignin on cellulose digestion. The miniaturized and semi-automated method we developed was usable at the laboratory scale and was reliable for mimicking the saccharification at the pilot scale using a steam explosion pretreatment and enzymatic hydrolysis. Therefore, this miniaturized method will allow the reliable screening of many genotypes for saccharification potential. These findings provide valuable information and tools for breeders to create genotypes combining high yield, suitable biomass composition, and high saccharification

Acid pretreatment and enzymatic saccharification of brown seaweed for polyhydroxybutyrate (PHB) production using Cupriavidus necator.

Science.gov (United States)

Azizi, Nahid; Najafpour, Ghasem; Younesi, Habibollah

2017-08-01

The brown seaweed Sargassum sp. was used as a feedstock to produce polyhydroxybutyarte (PHB) using Cupriavidus necator PTCC 1615. In order to release monomeric sugars, dilute acid hydrolysis of Sargassum sp. biomass was followed by enzymatic saccharification. In addition, the effect of different nitrogen sources was evaluated for PHB production. The fermentation of hydrolysate with the ammonium sulfate as selected nitrogen source resulted PHB yield of 0.54±0.01g/g reducing sugar. Then, NaCl was used as external stress factor which was added to the media. Addition of 8g/L NaCl had a positive impact on high PHB yield of 0.74±0.01g/g reducing sugar. Increasing trend of NaCl concentration to 16g/L was found to inhibit the production of PHB. Based on obtained results using 20g/L of reducing sugar, at desired condition the highest cell dry weight and PHB concentrations were 5.36±0.22 and 3.93±0.24g/L, respectively. The findings of this study reveal that Sargassum sp. is a promising feedstock for biopolymer production. The characteristics of produced PHB were analyzed by FTIR, differential scanning calorimetry and 1 H NMR. Copyright © 2017 Elsevier B.V. All rights reserved.

Increase in cellulose accumulation and improvement of saccharification by overexpression of arabinofuranosidase in rice.

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Minako Sumiyoshi

Full Text Available Cellulosic biomass is available for the production of biofuel, with saccharification of the cell wall being a key process. We investigated whether alteration of arabinoxylan, a major hemicellulose in monocots, causes an increase in saccharification efficiency. Arabinoxylans have β-1,4-D-xylopyranosyl backbones and 1,3- or 1,4-α-l-arabinofuranosyl residues linked to O-2 and/or O-3 of xylopyranosyl residues as side chains. Arabinose side chains interrupt the hydrogen bond between arabinoxylan and cellulose and carry an ester-linked feruloyl substituent. Arabinose side chains are the base point for diferuloyl cross-links and lignification. We analyzed rice plants overexpressing arabinofuranosidase (ARAF to study the role of arabinose residues in the cell wall and their effects on saccharification. Arabinose content in the cell wall of transgenic rice plants overexpressing individual ARAF full-length cDNA (OsARAF1-FOX and OsARAF3-FOX decreased 25% and 20% compared to the control and the amount of glucose increased by 28.2% and 34.2%, respectively. We studied modifications of cell wall polysaccharides at the cellular level by comparing histochemical cellulose staining patterns and immunolocalization patterns using antibodies raised against α-(1,5-linked l-Ara (LM6 and β-(1,4-linked d-Xyl (LM10 and LM11 residues. However, they showed no visible phenotype. Our results suggest that the balance between arabinoxylan and cellulose might maintain the cell wall network. Moreover, ARAF overexpression in rice effectively leads to an increase in cellulose accumulation and saccharification efficiency, which can be used to produce bioethanol.

Effect of γ-radiation on the saccharification of cellulose

International Nuclear Information System (INIS)

De la Rosa, A.M.; Banzon, R.B.; Abad, L.V.; Nuguid, Z.F.; Bulos, A.S.

1985-01-01

The effect of gamma radiation on the acid and saccharification of agricultural cellulosic wastes was investigated. Radiation doses of 200 KGy and higher significantly increased the saccharification of rice straw, rice hull and corn husk. The observed radiation effects varied with the cellulosic material. Rice straw exhibited the greatest radiosensitivity while rice hull showed the least susceptibility to gamma radiation. Possible mechanisms for the radiation-induced degradation of cellulose and agricultural cellulosic wastes are discussed. (author)

Simultaneous production of bio-ethanol and bleached pulp from red algae.

Science.gov (United States)

Yoon, Min Ho; Lee, Yoon Woo; Lee, Chun Han; Seo, Yung Bum

2012-12-01

The red algae, Gelidium corneum, was used to produce bleached pulp for papermaking and ethanol. Aqueous extracts obtained at 100-140 °C were subjected to saccharification, purification, fermentation, and distillation to produce ethanol. The solid remnants were bleached with chlorine dioxide and peroxide to make pulp. In the extraction process, sulfuric acid and sodium thiosulfate were added to increase the extract yield and to improve de-polymerization of the extracts, as well as to generate high-quality pulp. An extraction process incorporating 5% sodium thiosulfate by dry weight of the algae provided optimal production conditions for the production of both strong pulp and a high ethanol yield. These results suggest that it might be possible to utilize algae instead of trees and starch for pulp and ethanol production, respectively. Copyright © 2012 Elsevier Ltd. All rights reserved.

One-step process of hydrothermal and alkaline treatment of wheat straw for improving the enzymatic saccharification.

Science.gov (United States)

Sun, Shaolong; Zhang, Lidan; Liu, Fang; Fan, Xiaolin; Sun, Run-Cang

2018-01-01

To increase the production of bioethanol, a two-step process based on hydrothermal and dilute alkaline treatment was applied to reduce the natural resistance of biomass. However, the process required a large amount of water and a long operation time due to the solid/liquid separation before the alkaline treatment, which led to decrease the pure economic profit for production of bioethanol. Therefore, four one-step processes based on order of hydrothermal and alkaline treatment have been developed to enhance concentration of glucose of wheat straw by enzymatic saccharification. The aim of the present study was to systematically evaluated effect for different one-step processes by analyzing the physicochemical properties (composition, structural change, crystallinity, surface morphology, and BET surface area) and enzymatic saccharification of the treated substrates. In this study, hemicelluloses and lignins were removed from wheat straw and the morphologic structures were destroyed to various extents during the four one-step processes, which were favorable for cellulase absorption on cellulose. A positive correlation was also observed between the crystallinity and enzymatic saccharification rate of the substrate under the conditions given. The surface area of the substrate was positively related to the concentration of glucose in this study. As compared to the control (3.0 g/L) and treated substrates (11.2-14.6 g/L) obtained by the other three one-step processes, the substrate treated by one-step process based on successively hydrothermal and alkaline treatment had a maximum glucose concentration of 18.6 g/L, which was due to the high cellulose concentration and surface area for the substrate, accompanying with removal of large amounts of lignins and hemicelluloses. The present study demonstrated that the order of hydrothermal and alkaline treatment had significant effects on the physicochemical properties and enzymatic saccharification of wheat straw. The one

Same-vessel enzymatic saccharification and fermentation of organosolv/H2O2 pretreated oil palm (Elaeis guineensis Jacq.) fronds for bioethanol production: Optimization of process parameters

International Nuclear Information System (INIS)

Ofori-Boateng, Cynthia; Lee, Keat Teong

2014-01-01

Highlights: • Same vessel enzymatic saccharification and fermentation (SVSF) of pretreated OPFs. • Optimum conditions:37 °C, 8.0% solid loading, 14.0 g/l yeast concentration, pH 5.3. • Optimum bioethanol concentration and yield of 21.96 g/l and 84.65% respectively. • Organosolv/H 2 O 2 pretreatment of OPFs improved SVSF yield at high solid loading. - Abstract: Based on optimized pretreatment process, oil palm fronds (OPFs) were sequentially pretreated with 1.4% (w/v) aq. NaOH in 80% ethanol with ultrasound assistance (at 75 °C for 30 min) and 3% (v/v) aq. H 2 O 2 . Using the Box–Behnken design (BBD) of response surface methodology (RSM), bioethanol production from the sono-assisted organosolv/H 2 O 2 OPFs were optimized using same-vessel enzymatic saccharification and fermentation (SVSF) where both the hydrolysis and fermentation processes were carried out in one vessel simultaneously. Throughout the SVSF process, the incubation time and enzyme loading were kept at 72 h and 15 filter paper unit (FPU)/g substrate respectively. The other SVSF parameters which affect bioethanol yield such as temperature (X 1 : 30–50 °C), solid loading (X 2 : 5.0–10.0% w/v), yeast concentration (X 3 : 5.0–20 g/l) and pH (X 4 : 4.0–7.0) were optimized. Well fitted regression equations (R 2 > 0.97) obtained were able to predict reliable optimum bioethanol concentration and yield. The predicted optimum bioethanol concentration (i.e., 20.61 g/l) and yield (i.e., 84.60%) were attained at 36.94 °C (∼37 °C), 7.57% w/v solid loading (∼8.0% w/v), 13.97 g/l yeast concentration (∼14.0 g/l) and pH of 5.29 (∼5.30). Validated results indicated a maximum ethanol concentration and yield of 21.96 g/l and 84.65% respectively, which were closer to the predicted optimum responses. Using the optimum conditions, the highest bioethanol productivity of 0.76 g/l/h was observed at 12 h of SVSF process

Saccharification of cellulose by acetolysis

Energy Technology Data Exchange (ETDEWEB)

Tanaka, T; Yamanaka, S; Takinami, K

1978-01-01

For saccharification of cellulose, an acetolysis method using assimilable acid with a microorganism was applied. Based on this method, a new method which gave totally assimilable products was established. The rigid crystalline structure of cellulose was disrupted by acetolysis with 2-2.5 times as much acetic anhydride as cellulose on a weight basis and 1 N sulfuric acid as a catalyst. Then for cleavage of O-acetyl ester and glycosidic bonds, the resulting amorphous acetolysate of cellulose could easily be hydrolyzed by heating in 1 N sulfuric acid at 120/sup 0/C for 1-1.5 h without over-disruption of glucose. Ninety-eight % of the cellulose used was recovered in the form of hydrolysate having about 30% saccharide concentration. The hydrolysate obtained was composed of 74% glucose, 13% cellobiose and 11% mono-O-acetyl glucose on a weight basis.

Scale up of ethanol production using pulp mill wastewater sludge by cellulase and saccharomyces cerevisiae

International Nuclear Information System (INIS)

Kunchada Sangasintu; Petchporn Chawakitchareon

2010-01-01

This study aimed to evaluate the potential use of pulp mill wastewater sludge as substrate in ethanol production. The simultaneous saccharification and fermentation process was conducted by using Saccharomyces cerevisiae TISTR 5339 under optimum proportion of cellulase and pulp mill wastewater sludge. The ethanol production from cellulosic materials in simultaneous saccharification and fermentation needs cooperation between cellulase and yeast. The cellulase hydrolyzes cellulose to sugar while yeast utilizes sugar to produce ethanol. The pulp mill wastewater sludge has an average content of 73.3 % hemi cellulose, 67.1 % alpha cellulose, 4.7 % beta cellulose and 1.4 % gamma cellulose. The experimental results indicated that the volume of the ethanol tend to increase with time, providing the maximum ethanol yield of 0.69 g/g on the 7"t"h day, the last day of the experiment. The ethanol production was scaled up in 5 L fermentor under optimum proportion and increased the fermentation period. It was found that the ethanol production gave the maximum ethanol yield of 1.14 g/g on the 9"t"h day of the totally 13 days experimentation. These results showed that the cellulose from pulp mill wastewater sludge was as effective substrate for ethanol production and alternative energy for the future. (author)

Enhancement of enzymatic saccharification of Eucalyptus globulus: steam explosion versus steam treatment.

Science.gov (United States)

Martin-Sampedro, Raquel; Revilla, Esteban; Villar, Juan C; Eugenio, Maria E

2014-09-01

Steam explosion and steam pre-treatment have proved capable of enhancing enzymatic saccharification of lignocellulosic materials. However, until now, these methods had not been compared under the same operational conditions and using the same raw material. Both pre-treatments lead to increased yields in the saccharification of Eucalyptus globulus; but results have been better with steam pre-treatments, despite the more accessible surface of exploded samples. The reason for this finding could be enzymatic inhibition: steam explosion causes a more extensive extraction of hemicelluloses and releases a greater amount of degradation products which can inhibit enzymatic action. Enzymatic inhibition is also dependent on the amount and chemical structure of lignin, which was also a contributing factor to the lower enzymatic yields obtained with the most severe pre-treatment. Thus, the highest yields (46.7% glucose and 73.4% xylose yields) were obtained after two cycle of steam treatment, of 5 and 3 min, at 183°C. Copyright © 2014 Elsevier Ltd. All rights reserved.

Optimization of Saccharification Conditions of Lignocellulosic Biomass under Alkaline Pre-Treatment and Enzymatic Hydrolysis

Directory of Open Access Journals (Sweden)

Rafał Łukajtis

2018-04-01

Full Text Available Pre-treatment is a significant step in the production of second-generation biofuels from waste lignocellulosic materials. Obtaining biofuels as a result of fermentation processes requires appropriate pre-treatment conditions ensuring the highest possible degree of saccharification of the feed material. An influence of the following process parameters were investigated for alkaline pre-treatment of Salix viminalis L.: catalyst concentration (NaOH, temperature, pre-treatment time and granulation. For this purpose, experiments were carried out in accordance to the Box-Behnken design for four factors. In the saccharification process of the pre-treated biomass, cellulolytic enzymes immobilized on diatomaceous earth were used. Based on the obtained results, a mathematical model for the optimal conditions of alkaline pre-treatment prediction is proposed. The optimal conditions of alkaline pre-treatment are established as follows: granulation 0.75 mm, catalyst concentration 7%, pre-treatment time 6 h and temperature 65 °C if the saccharification efficiency and cost analysis are considered. An influence of the optimized pre-treatment on both the chemical composition and structural changes for six various lignocellulosic materials (energetic willow, energetic poplar, beech, triticale, meadow grass, corncobs was investigated. SEM images of raw and pre-treated biomass samples are included in order to follow the changes in the biomass structure during hydrolysis.

Saccharification with Phanerochaete chrysosporium and Pleurotus ...

African Journals Online (AJOL)

Lignocellulosic biomass has a great potential as raw material for second and third generation biofuels since it is the most abundant carbohydrate on earth and the main component of agricultural waste; however, saccharification of lignocellulosic biomass is crucial for the establishment of a carbohydratebased economy.

Enzymatic saccharification of liquid hot water and dilute sulfuric acid pretreated oil palm empty fruit bunch and sugarcane bagasse

Science.gov (United States)

Risanto, L.; Fitria; Fajriutami, T.; Hermiati, E.

2018-03-01

Oil palm empty fruit bunch (OPEFB) and sugarcane bagasse (SB) are potential feedstocks for the production of bioethanol. In this study OPEFB and SB were pretreated by liquid hot water and dilute sulfuric acid (3% H2SO4), and continued with enzymatic saccharification. Heating treatment for both methods was conducted in an autoclave at 121 °C for 1 hr. The saccharification was performed up to 72 hours with cellulase enzyme loading of 10, 20, and 30 FPU per g biomass. Results showed that OPEFB and SB pretreated with H2SO4 produced higher reducing sugars than those pretreated by liquid hot water. Higher enzyme loading also resulted in higher reducing sugars. Reducing sugars obtained from enzymatic saccharification of OPEFB were higher than those obtained from SB. The highest total reducing sugars (50.48 g/100 g biomass) was obtained from OPEFB pretreated with 3% H2SO4 at enzyme loading of 30 FPU per g biomass.

Range of cell-wall alterations enhance saccharification in Brachypodium distachyon mutants

DEFF Research Database (Denmark)

Marriott, Poppy E; Sibout, Richard; Lapierre, Catherine

2014-01-01

saccharification with an industrial polysaccharide-degrading enzyme mixture. From an initial screen of 2,400 M2 plants, we selected 12 lines that showed heritable improvements in saccharification, mostly with no significant reduction in plant size or stem strength. Characterization of these putative mutants...

Liquid nitrogen pretreatment of eucalyptus sawdust and rice hull for enhanced enzymatic saccharification.

Science.gov (United States)

Castoldi, Rafael; Correa, Vanesa G; de Morais, Gutierrez Rodrigues; de Souza, Cristina G M; Bracht, Adelar; Peralta, Rosely A; Peralta-Muniz Moreira, Regina F; Peralta, Rosane M

2017-01-01

In this work, liquid nitrogen was used for the first time in the pretreatment of plant biomasses for purposes of enzymatic saccharification. After treatment (cryocrushing), the initial rates of the enzymatic hydrolysis of eucalyptus sawdust and rice hull were increased more than ten-fold. Cryocrushing did not modify significantly the contents of cellulose, hemicellulose and lignin in both eucalyptus sawdust and rice hulls. However, substantial disorganization of the lignocellulosic materials in consequence of the pretreatment could be observed by electron microscopy. Cryocrushing was highly efficient in improving the saccharification of the holocellulose component of the plant biomasses (from 4.3% to 54.1% for eucalyptus sawdust and from 3.9% to 40.6% for rice hull). It is important to emphasize that it consists in a simple operation with low requirements of water and chemicals, no corrosion, no release of products such as soluble phenolics, furfural and hydroxymethylfurfural and no waste generation. Copyright © 2016 Elsevier Ltd. All rights reserved.

Sequential enzymatic saccharification and fermentation of ionic liquid and organosolv pretreated agave bagasse for ethanol production

Energy Technology Data Exchange (ETDEWEB)

Pérez-Pimienta, Jose A. [Univ. Autonoma de Nayarit, Tepic (Mexico); Vargas-Tah, Alejandra [Univ. Nacional Autonoma de Mexico (UNAM), Cuernavaca (Mexico).; López-Ortega, Karla M. [Univ. Autonoma de Nayarit, Tepic (Mexico); Medina-López, Yessenia N. [Univ. Autonoma de Nayarit, Tepic (Mexico); Mendoza-Pérez, Jorge A. [Inst. Politecnico Nacional (IPN), Mexico City (Mexico); Avila, Sayeny [Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Singh, Seema [Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Sandia National Lab. (SNL-CA), Livermore, CA (United States); Simmons, Blake A. [Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Sandia National Lab. (SNL-CA), Livermore, CA (United States); Loaces, Inés [Univ. Nacional Autonoma de Mexico (UNAM), Cuernavaca (Mexico).; Martinez, Alfredo [Univ. Nacional Autonoma de Mexico (UNAM), Cuernavaca (Mexico).

2016-11-16

Agave bagasse (AGB) has gained recognition as a drought-tolerant biofuel feedstock with high productivity in semiarid regions. A comparative analysis of ionic liquid (IL) and organosolv (OV) pretreatment technologies in AGB was performed using a sequential enzymatic saccharification and fermentation (SESF) strategy with cellulolytic enzymes and the ethanologenic Escherichia coli strain MS04. After pretreatment, 86% of xylan and 45% of lignin were removed from OV-AGB, whereas IL-AGB reduced lignin content by 28% and xylan by 50% when compared to the untreated biomass. High glucan ( > 90%) and xylan ( > 83%) conversion was obtained with both pretreated samples. During the fermentation stage (48 h), 12.1 and 12.7 kg of ethanol were produced per 100 kg of untreated AGB for IL and OV, respectively. These comparative analyses showed the advantages of SESF using IL and OV in a biorefinery configuration where a better understanding of AGB recalcitrance is key for future applications.

Incorporation of flavonoid derivatives or pentagalloyl glucose into lignin enhances cell wall saccharification following mild alkaline or acidic pretreatments

Science.gov (United States)

Partial substitution of normal monolignols with phenolic precursors from other metabolic pathways may improve the susceptibility of lignified biomass to chemical pretreatment and enzymatic saccharification for biofuel production. Flavonoids and gallate esters readily undergo oxidative coupling react...

Saccharification of ozonated sugarcane bagasse using enzymes from Myceliophthora thermophila JCP 1-4 for sugars release and ethanol production.

Science.gov (United States)

de Cassia Pereira, Josiani; Travaini, Rodolfo; Paganini Marques, Natalia; Bolado-Rodríguez, Silvia; Bocchini Martins, Daniela Alonso

2016-03-01

The saccharification of ozonated sugarcane bagasse (SCB) by enzymes from Myceliophthora thermophila JCP 1-4 was studied. Fungal enzymes provided slightly higher sugar release than commercial enzymes, working at 50°C. Sugar release increased with temperature increase. Kinetic studies showed remarkable glucose release (4.99 g/L, 3%w/w dry matter) at 60°C, 8 h of hydrolysis, using an enzyme load of 10 FPU (filter paper unit). FPase and β-glucosidase activities increased during saccharification (284% and 270%, respectively). No further significant improvement on glucose release was observed increasing the enzyme load above 7.5 FPU per g of cellulose. Higher dry matter contents increased sugars release, but not yields. The fermentation of hydrolysates by Saccharomyces cerevisiae provided glucose-to-ethanol conversions around to 63%. Copyright © 2015 Elsevier Ltd. All rights reserved.

Saccharification of gamma-ray and alkali pretreated lignocellulosics

International Nuclear Information System (INIS)

Begum, A.; Choudhury, N.

1988-01-01

Enzymic saccharification of gamma ray and alkali pretreated sawdust, rice straw, and sugar cane bagasse showed higher release of reducing sugar from pretreated substrates. By gamma ray treatment alone (500 kGy) reducing sugar release of 2.8, 9.2, and 10 g/l was obtained from 7.5% (w/v) sawdust, rice straw, and bagasse and the same substrates showed reducing sugar release of 4.2, 30, and 20 g/l respectively when treated with alkali (0.1 g/g). Combination of gamma ray with alkali treatment further increased the reducing sugar release to 10.2, 33, and 36 g/l from sawdust, rice straw, and bagasse respectively. The effects of gamma ray and alkali treatment on saccharification varied with the nature of the substrate

Process Design Report for Wood Feedstock: Lignocellulosic Biomass to Ethanol Process Desing and Economics Utilizing Co-Current Dilute Acid Prehydrolysis and Enzymatic Hydrolysis Current and Futuristic Scenarios

Energy Technology Data Exchange (ETDEWEB)

Wooley, Robert [National Renewable Energy Lab. (NREL), Golden, CO (United States); Ruth, Mark [National Renewable Energy Lab. (NREL), Golden, CO (United States); Sheehan, John [National Renewable Energy Lab. (NREL), Golden, CO (United States); Ibsen, Kelly [National Renewable Energy Lab. (NREL), Golden, CO (United States); Majdeski, Henry [Delta-T Corporation, Lexington, KY (United States); Galves, Adrian [Delta-T Corporation, Lexington, KY (United States)

1999-07-01

The National Renewable Energy Laboratory (NREL) has undertaken a complete review and update of the process design and economic model for the biomass-to-ethanol process based on co-current dilute acid prehydrolysis, along with simultaneous saccharification (enzymatic) and co-fermentation. The process design includes the core technologies being researched by the U.S. Department of Energy (DOE): prehydrolysis, simultaneous saccharification and co-fermentation, and cellulase enzyme production.

Enzymatic Saccharification of Lignocelluloses Should be Conducted at Elevated pH 5.2-6.2

Science.gov (United States)

T.Q. Lan; Hongming Lou; J.Y. Zhu

2013-01-01

This study revealed that cellulose enzymatic saccharification response curves of lignocellulosic substrates were very different from those of pure cellulosic substrates in terms of optimal pH and pH operating window. The maximal enzymatic cellulose saccharification of lignocellulosic substrates occurs at substrate suspension

A review of conversion processes for bioethanol production with a focus on syngas fermentation

Directory of Open Access Journals (Sweden)

Mamatha Devarapalli

2015-09-01

Full Text Available Bioethanol production from corn is a well-established technology. However, emphasis on exploring non-food based feedstocks is intensified due to dispute over utilization of food based feedstocks to generate bioethanol. Chemical and biological conversion technologies for non-food based biomass feedstocks to biofuels have been developed. First generation bioethanol was produced from sugar based feedstocks such as corn and sugar cane. Availability of alternative feedstocks such as lignocellulosic and algal biomass and technology advancement led to the development of complex biological conversion processes, such as separate hydrolysis and fermentation (SHF, simultaneous saccharification and fermentation (SSF, simultaneous saccharification and co-fermentation (SSCF, consolidated bioprocessing (CBP, and syngas fermentation. SHF, SSF, SSCF, and CBP are direct fermentation processes in which biomass feedstocks are pretreated, hydrolyzed and then fermented into ethanol. Conversely, ethanol from syngas fermentation is an indirect fermentation that utilizes gaseous substrates (mixture of CO, CO2 and H2 made from industrial flue gases or gasification of biomass, coal or municipal solid waste. This review article provides an overview of the various biological processes for ethanol production from sugar, lignocellulosic, and algal biomass. This paper also provides a detailed insight on process development, bioreactor design, and advances and future directions in syngas fermentation.

Ethanol production from kitchen waste using the flocculating yeast Saccharomyces cerevisiae strain KF-7

International Nuclear Information System (INIS)

Tang, Y.-Q.; Koike, Yoji; Liu Kai; An, M.-Z.; Morimura, Shigeru; Wu Xiaolei; Kida, Kenji

2008-01-01

A process for producing ethanol from kitchen waste was developed in this study. The process consists of freshness preservation of the waste, saccharification of the sugars in the waste, continuous ethanol fermentation of the saccharified liquid, and anaerobic treatment of the saccharification residue and the stillage. Spraying lactic acid bacteria (LCB) on the kitchen waste kept the waste fresh for over 1 week. High glucose recovery (85.5%) from LCB-sprayed waste was achieved after saccharification using Nagase N-40 glucoamylase. The resulting saccharified liquid was used directly for ethanol fermentation, without the addition of any nutrients. High ethanol productivity (24.0 g l -1 h -1 ) was obtained when the flocculating yeast strain KF-7 was used in a continuous ethanol fermentation process at a dilution rate of 0.8 h -1 . The saccharification residue was mixed with stillage and treated in a thermophilic anaerobic continuous stirred tank reactor (CSTR); a VTS loading rate of 6 g l -1 d -1 with 72% VTS digestion efficiency was achieved. Using this process, 30.9 g ethanol, and 65.2 l biogas with 50% methane, was produced from 1 kg of kitchen waste containing 118.0 g total sugar. Thus, energy in kitchen waste can be converted to ethanol and methane, which can then be used as fuels, while simultaneously treating kitchen waste

Ethanol production from kitchen waste using the flocculating yeast Saccharomyces cerevisiae strain KF-7

Energy Technology Data Exchange (ETDEWEB)

Tang, Yue-Qin; Liu, Kai; An, Ming-Zhe; Morimura, Shigeru; Kida, Kenji [Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555 (Japan); Koike, Yoji [Tokyo Gas Co., Ltd., 1-7-7 Suehiro-cho, Tsurumi-ku, Yokohama City, Kanagawa 230-0045 (Japan); Wu, Xiao-Lei [Department of Energy and Resources Engineering, College of Engineering, Peking University, Beijing 100871 (China)

2008-11-15

A process for producing ethanol from kitchen waste was developed in this study. The process consists of freshness preservation of the waste, saccharification of the sugars in the waste, continuous ethanol fermentation of the saccharified liquid, and anaerobic treatment of the saccharification residue and the stillage. Spraying lactic acid bacteria (LCB) on the kitchen waste kept the waste fresh for over 1 week. High glucose recovery (85.5%) from LCB-sprayed waste was achieved after saccharification using Nagase N-40 glucoamylase. The resulting saccharified liquid was used directly for ethanol fermentation, without the addition of any nutrients. High ethanol productivity (24.0 g l{sup -1} h{sup -1}) was obtained when the flocculating yeast strain KF-7 was used in a continuous ethanol fermentation process at a dilution rate of 0.8 h{sup -1}. The saccharification residue was mixed with stillage and treated in a thermophilic anaerobic continuous stirred tank reactor (CSTR); a VTS loading rate of 6 g l{sup -1} d{sup -1} with 72% VTS digestion efficiency was achieved. Using this process, 30.9 g ethanol, and 65.2 l biogas with 50% methane, was produced from 1 kg of kitchen waste containing 118.0 g total sugar. Thus, energy in kitchen waste can be converted to ethanol and methane, which can then be used as fuels, while simultaneously treating kitchen waste. (author)

Strategies for enzyme saving during saccharification of pretreated lignocellulo-starch biomass: effect of enzyme dosage and detoxification chemicals

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M.G. Mithra

2017-08-01

Full Text Available Two strategies leading to enzyme saving during saccharification of pretreated lignocellulo-starch biomass (LCSB was investigated which included reducing enzyme dosage by varying their levels in enzyme cocktails and enhancing the fermentable sugar yield in enzyme-reduced systems using detoxification chemicals. Time course release of reducing sugars (RS during 24–120 h was significantly higher when an enzyme cocktail containing full dose of cellulase (16 FPU/g cellulose along with half dose each of xylanase (1.5 mg protein/g hemicelluloses and Stargen (12.5 μl/g biomass was used to saccharify conventional dilute sulphuric acid (DSA pretreated biomass compared to a parallel system where only one-fourth the dose of the latter two enzymes was used. The reduction in RS content in the 120 h saccharified mash to the extent of 3–4 g/L compared to the system saccharified with full complement of the three enzymes could be overcome considerably by supplementing the system (half dose of two enzymes with detoxification chemical mix incorporating Tween 20, PEG 4000 and sodium borohydride. Microwave (MW-assisted DSA pretreated biomass on saccharification with enzyme cocktail having full dose of cellulase and half dose of Stargen along with detoxification chemicals gave significantly higher RS yield than DSA pretreated system saccharified using three enzymes. The study showed that xylanase could be eliminated during saccharification of MW-assisted DSA pretreated biomass without affecting RS yield when detoxification chemicals were also supplemented. The Saccharification Efficiency and Overall Conversion Efficiency were also high for the MW-assisted DSA pretreated biomass. Since whole slurry saccharifcation of pretreated biomass is essential to conserve fermentable sugars in LCSB saccharification, detoxification of soluble inhibitors is equally important as channelling out of insoluble lignin remaining in the residue. As one of the major factors contributing

Effect of pH on simultaneous saccharification and isomerization by glucoamylase and glucose isomerase.

Science.gov (United States)

Mishra, Abha; Debnath Das, Meera

2002-01-01

pH and temperature play critical roles in multistep enzymatic conversions. In such conversions, the optimal pH for individual steps differs greatly. In this article, we describe the production of glucoamylase (from Aspergillus oryzae MTCC152 in solid-state fermentation) and glucose isomerase (from Streptomyces griseus NCIM2020 in submerged fermentation), used in industries for producing high-fructose syrup. Optimum pH for glucoamylase was found to be 5.0. For glucose isomerase, the optimum pH ranged between 7.0 and 8.5, depending on the type of buffer used. Optimum temperature for glucoamylase and glucose isomerase was 50 and 60 degrees C, respectively. When both the enzymatic conversions were performed simultaneously at a compromised pH of 6.5, both the enzymes showed lowered activity. We also studied the kinetics at different pHs, which allows the two-step reaction to take place simultaneously. This was done by separating two steps by a thin layer of urease. Ammonia generated by the hydrolysis of urea consumed the hydrogen ions, thereby allowing optimal activity of glucose isomerase at an acidic pH of 5.0.

Pyrolysis of wastes generated through saccharification of oak tree by using CO2 as reaction medium

International Nuclear Information System (INIS)

Kim, Jieun; Lee, Jechan; Kim, Ki-Hyun; Ok, Yong Sik; Jeon, Young Jae; Kwon, Eilhann E.

2017-01-01

Highlights: • Potential utilization of biomass waste generated from bioethanol production. • Enhanced generation of syngas from pyrolysis of oak tree waste by using CO 2 . • Reduction of tar formation in pyrolysis of oak tree waste. • Modification of morphology of oak tree waste biochar by using CO 2 in pyrolysis. - Abstract: In this study, the production of bioethanol was evaluated through a series of saccharification and fermentation of lignocellulosic biomass (e.g., oak tree) pre-treated with H 2 SO 4 , NH 3 , or NaOH using a yeast (Pichia stipitis). In addition, it was investigated the effects of CO 2 on pyrolysis of the biomass wastes remaining after saccharification of the three pre-treated oak tree (BWs: BW-H 2 SO 4 , BW-NH 3 , and BW-NaOH). Thus, this work emphasizes the mechanistic understanding of CO 2 in pyrolysis of BWs. The effect of CO 2 was most noticeable in syngas, as the ratio of CO and H 2 exhibited a 20 to 30-fold increase at >550 °C. The CO/H 2 ratio of pyrolysis of the waste in CO 2 is ∼1100% of that of pyrolysis of the waste in N 2 at 720 °C. Such proliferation of syngas led to the subsequent reduction of tar since the substantial amount of tar was consumed as a precursor of syngas: CO 2 not only expedited the thermal cracking of volatile organic compounds (VOCs), but also reacted with those VOCs. The morphologic modification of biochars also occurred in the presence of CO 2 via heterogeneous reaction between CO 2 and surface of BWs. In summary, this study shows a utilization of an oak tree waste generated from saccharification for bioethanol production as a pyrolysis feedstock to recover energy (i.e., syngas production). The use of CO 2 as pyrolysis medium not only enhanced syngas production from oak tree waste but also reduced tar formation by thermal decomposition of VOCs and reaction between VOCs and CO 2 . The process shown in this study can be used as a potential high energy recovery from a biomass waste by utilizing potent

Effect of 60Co γ-irradiation on saccharification of uncooked sweet potato material

International Nuclear Information System (INIS)

Hu Tingchun; Xiong Xingyao; Yi Jinqiong; Wang Keqin; Su Xiaojun; Zou Jianfeng

2010-01-01

Using the starch and powder of sweet potato of Xiangshu 86 and Xiangshu 541 as materials, the effect of 60 Co γ-irradiation on the structure of starch particle and the efficiency of saccharification were studied. The result showed that some reticulate flaws appeared in the surface of irradiated starch particles, and the reticulate flaws were increased with the increase of irradiation dose. The content of reducing sugar and total soluble sugar in both starch and the powder were obviously increased along with the increase of irradiation dose ranged from 50 to 1200 kGy. The saccharification efficiency of Xiangshu 86 and Xiangshu 541 was obviously difference at the dose lower than 500 kGy, and then the efficiency showed the similar trends at higher dose irradiation, the saccharification rate reached the highest value after the treatment of 1200 kGy irradiation. (authors)

Construction of a Bacterial Cellulase Cocktail for Saccharification of Regenerated Cellulose and Pretreated Corn Stover

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Alei Geng

2015-09-01

Full Text Available To apply bacterial cellulases for efficient saccharification of biomass, three Clostridium thermocellum cellulases and a Thermoanaerobacter brockii β-1,4-glucosidase were synthesized in Escherichia coli, and the proportions among them were optimized. When the activities of CelD, CBHA, CBH48Y, and CglT were set at 554, 0.91, 0.91, and 856 mU per assay, respectively, the percent conversion of regenerated cellulose (0.92 g/L reached 80.9% within 24 h at 60 °C without shaking. Meanwhile, the percent conversion of pretreated corn stover (0.62 g/L reached 70.1%. Gradually raising the loads of regenerated cellulose from 0.92 to 4.58 g/L resulted in a linear increase in glucose production from 870 to 3208 μg (R2=0.997, as well as a decrease in the percent conversion from 80.9% to 59.6%. These findings suggested that the cellulase cocktail is efficient in saccharification of regenerated cellulose, as well as pretreated corn stover, and has potential applications in the biofuels industry.

A Shortcut to the Production of High Ethanol Concentration from Jerusalem Artichoke Tubers

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Wei-Guo Zhang

2005-01-01

Full Text Available Aspergillus niger SL-09, a newly isolated exoinulinase-hyperproducing strain, and Saccharomyces cerevisiae Z-06, with high ethanol tolerance, were used in a fed-batch process for simultaneous saccharification and fermentation of Jerusalem artichoke tuber mash and flour. S. cerevisiae Z-06 utilized 98 % of the total sugar and produced 19.6 % of ethanol in 48 h. In this process the conversion efficiency of the fermentation of Jerusalem artichoke and the production of ethanol were 90 % of the theoretical ethanol yield and the cost of the production of flour was cut nearly into half.

Heterologous expression of Pycnoporus cinnabarinus cellobiose dehydrogenase in Pichia pastoris and involvement in saccharification processes

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Bey Mathieu

2011-12-01

Full Text Available Abstract Background Cellobiose dehydrogenase (CDH is an extracellular hemoflavoenzyme produced by lignocellulose-degrading fungi including Pycnoporus cinnabarinus. We investigated the cellulolytic system of P. cinnabarinus, focusing on the involvement of CDH in the deconstruction of lignocellulosic biomass. Results First, P. cinnabarinus growth conditions were optimized for CDH production. Following growth under cellulolytic conditions, the main components secreted were cellulases, xylanases and CDH. To investigate the contribution of P. cinnabarinus secretome in saccharification processes, the Trichoderma reesei enzymatic cocktail was supplemented with the P. cinnabarinus secretome. A significant enhancement of the degradation of wheat straw was observed with (i the production of a large amount of gluconic acid, (ii increased hemicellulose degradation, and (iii increased overall degradation of the lignocellulosic material. P. cinnabarinus CDH was heterologously expressed in Pichia pastoris to obtain large amounts of pure enzyme. In a bioreactor, the recombinant CDH (rCDH expression level reached 7800 U/L. rCDH exhibited values of biochemical parameters similar to those of the natural enzyme, and was able to bind cellulose despite the absence of a carbohydrate-binding module (CBM. Following supplementation of purified rCDH to T. reesei enzymatic cocktail, formation of gluconic acid and increased hemicellulose degradation were observed, thus confirming the previous results observed with P. cinnabarinus secretome. Conclusions We demonstrate that CDH offers an attractive tool for saccharification process enhancement due to gluconic acid production from raw lignocellulosic material.

Separate and Simultaneous enzymatic hydrolysis and fermentation of wheat hemicellulose with recombinant xylose utilizing Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Olsson, Lisbeth; Sørensen, H. R.; Dam, B. P

2006-01-01

Fermentations with three different xylose-utilizing recombinant Saccharomyces cerevisiae strains (F12, CR4, and CB4) were performed using two different wheat hemicellulose substrates, unfermented starch free fibers, and an industrial ethanol fermentation residue, vinasse. With CR4 and F12......, the maximum ethanol concentrations obtained were 4.3 and 4 g/L, respectively, but F12 converted xylose 15% faster than CR4 during the first 24 h. The comparison of separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) with F12 showed that the highest, maximum...... ethanol concentrations were obtained with SSF. In general, the volumetric ethanol productivity was initially, highest in the SHF, but the overall volumetric ethanol productivity ended up being maximal in the SSF, at 0.013 and 0.010 g/Lh, with starch free fibers and vinasse, respectively....

Ethanol production from paper sludge using Kluyveromyces marxianus

International Nuclear Information System (INIS)

Madrid, Lina Maria; Quintero Diaz, Juan Carlos

2011-01-01

Recycled paper sludge is a promising raw material for ethanol production. In this study, we first evaluated the effects of ethanol concentration, solids load, and cellulose crystallinity on the enzymatic hydrolysis of cellulose to produce reducing sugars. We then evaluated the production of ethanol by either saccharification and simultaneous fermentation (SSF) or separated hydrolysis and fermentation (SHF) using the yeast Kluyveromyces marxianus ATCC 36907. We found that cellulose hydrolysis decreased as ethanol concentrations increased; at 40 g/L ethanol, the reducing sugar production was decreased by 79 %. Hydrolysis also decreased as solids load increased; at 9 % of solids, the cellulose conversion was 76 % of the stoichiometric production. The ethanol yield and cellulose conversion rate were higher with SSF as opposed to SHF processes at 72 h of treatment.

Laboratory-scale method for enzymatic saccharification of lignocellulosic biomass at high-solids loadings

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Dibble Clare J

2009-11-01

Full Text Available Abstract Background Screening new lignocellulosic biomass pretreatments and advanced enzyme systems at process relevant conditions is a key factor in the development of economically viable lignocellulosic ethanol. Shake flasks, the reaction vessel commonly used for screening enzymatic saccharifications of cellulosic biomass, do not provide adequate mixing at high-solids concentrations when shaking is not supplemented with hand mixing. Results We identified roller bottle reactors (RBRs as laboratory-scale reaction vessels that can provide adequate mixing for enzymatic saccharifications at high-solids biomass loadings without any additional hand mixing. Using the RBRs, we developed a method for screening both pretreated biomass and enzyme systems at process-relevant conditions. RBRs were shown to be scalable between 125 mL and 2 L. Results from enzymatic saccharifications of five biomass pretreatments of different severities and two enzyme preparations suggest that this system will work well for a variety of biomass substrates and enzyme systems. A study of intermittent mixing regimes suggests that mass transfer limitations of enzymatic saccharifications at high-solids loadings are significant but can be mitigated with a relatively low amount of mixing input. Conclusion Effective initial mixing to promote good enzyme distribution and continued, but not necessarily continuous, mixing is necessary in order to facilitate high biomass conversion rates. The simplicity and robustness of the bench-scale RBR system, combined with its ability to accommodate numerous reaction vessels, will be useful in screening new biomass pretreatments and advanced enzyme systems at high-solids loadings.

Potential of giant reed (Arundo donax L. for second generation ethanol production

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Claudia Fernanda Lemons e Silva

2015-01-01

Conclusions: The fermentability of the pretreated biomass was performed successfully through the conception of simultaneous saccharification and fermentation resulting in approximately 75 L of ethanol per ton of cellulose.

Comparison of the efficiency of bacterial and fungal laccases in delignification and detoxification of steam-pretreated lignocellulosic biomass for bioethanol production.

Science.gov (United States)

De La Torre, María; Martín-Sampedro, Raquel; Fillat, Úrsula; Eugenio, María E; Blánquez, Alba; Hernández, Manuel; Arias, María E; Ibarra, David

2017-11-01

This study evaluates the potential of a bacterial laccase from Streptomyces ipomoeae (SilA) for delignification and detoxification of steam-exploded wheat straw, in comparison with a commercial fungal laccase from Trametes villosa. When alkali extraction followed by SilA laccase treatment was applied to the water insoluble solids fraction, a slight reduction in lignin content was detected, and after a saccharification step, an increase in both glucose and xylose production (16 and 6%, respectively) was observed. These effects were not produced with T. villosa laccase. Concerning to the fermentation process, the treatment of the steam-exploded whole slurry with both laccases produced a decrease in the phenol content by up to 35 and 71% with bacterial and fungal laccases, respectively. The phenols reduction resulted in an improved performance of Saccharomyces cerevisiae during a simultaneous saccharification and fermentation (SSF) process, improving ethanol production rate. This enhancement was more marked with a presaccharification step prior to the SSF process.

Optimization of bioethanol production from simultaneous ...

African Journals Online (AJOL)

ADOWIE PERE

fermentation of pineapple peels using Saccharomyces cerevisiae ... ABSTRACT: In this study, bioethanol production from the simultaneous ... in turn has resulted in the need to find a source of ... fruit in the world after Banana and Citrus and.

Evaluation of Relationships between Growth Rate, Tree Size, Lignocellulose Composition, and Enzymatic Saccharification in Interspecific Corymbia Hybrids and Parental Taxa.

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Healey, Adam L; Lee, David J; Lupoi, Jason S; Papa, Gabriella; Guenther, Joel M; Corno, Luca; Adani, Fabrizio; Singh, Seema; Simmons, Blake A; Henry, Robert J

2016-01-01

In order for a lignocellulosic bioenergy feedstock to be considered sustainable, it must possess a high rate of growth to supply biomass for conversion. Despite the desirability of a fast growth rate for industrial application, it is unclear what effect growth rate has on biomass composition or saccharification. We characterized Klason lignin, glucan, and xylan content with response to growth in Corymbia interspecific F1 hybrid families (HF) and parental species Corymbia torelliana and C. citriodora subspecies variegata and measured the effects on enzymatic hydrolysis from hydrothermally pretreated biomass. Analysis of biomass composition within Corymbia populations found similar amounts of Klason lignin content (19.7-21.3%) among parental and hybrid populations, whereas glucan content was clearly distinguished within C. citriodora subspecies variegata (52%) and HF148 (60%) as compared to other populations (28-38%). Multiple linear regression indicates that biomass composition is significantly impacted by tree size measured at the same age, with Klason lignin content increasing with diameter breast height (DBH) (+0.12% per cm DBH increase), and glucan and xylan typically decreasing per DBH cm increase (-0.7 and -0.3%, respectively). Polysaccharide content within C. citriodora subspecies variegata and HF-148 were not significantly affected by tree size. High-throughput enzymatic saccharification of hydrothermally pretreated biomass found significant differences among Corymbia populations for total glucose production from biomass, with parental Corymbia torelliana and hybrids HF-148 and HF-51 generating the highest amounts of glucose (~180 mg/g biomass, respectively), with HF-51 undergoing the most efficient glucan-to-glucose conversion (74%). Based on growth rate, biomass composition, and further optimization of enzymatic saccharification yield, high production Corymbia hybrid trees are potentially suitable for fast-rotation bioenergy or biomaterial production.

Evaluation of relationships between growth rate, tree size, lignocellulose composition and enzymatic saccharification in interspecific Corymbia hybrids and parental taxa.

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Adam L Healey

2016-11-01

Full Text Available In order for a lignocellulosic bioenergy feedstock to be considered sustainable, it must possess a high rate of growth to supply biomass for conversion. Despite the desirability of a fast growth rate for industrial application, it is unclear what effect growth rate has on biomass composition or saccharification. We characterized Klason lignin, glucan, and xylan content with response to growth in Corymbia interspecific F1 hybrid families (HF and parental species C. torelliana (CT and C. citriodora subspecies variegata (CCV and measured the effects on enzymatic hydrolysis from hydrothermally pretreated biomass. Analysis of biomass composition within Corymbia populations found similar amounts of Klason lignin content (19.7-21.3% among parental and hybrid populations, whereas glucan content was clearly distinguished within CCV (52% and HF148 (60% as compared to other populations (28-38%. Multiple linear regression indicates that biomass composition is significantly impacted by tree size measured at the same age, with Klason lignin content increasing with diameter breast height (DBH (+0.12% per cm DBH increase, and glucan and xylan typically decreasing per DBH cm increase (-0.7% and -0.3%, respectively. Polysaccharide content within CCV and HF-148 were not significantly affected by tree size. High-throughput enzymatic saccharification of hydrothermally pretreated biomass found significant differences among Corymbia populations for total glucose production from biomass, with parental CT and hybrids HF-148 and HF-51 generating the highest amounts of glucose (~180 mg/g biomass, respectively, with HF-51 undergoing the most efficient glucan-to-glucose conversion (74%. Based on growth rate, biomass composition, and further optimization of enzymatic saccharification yield, high production Corymbia hybrid trees are potentially suitable for fast-rotation bioenergy or biomaterial production.

Epigallocatechin gallate incorporation into lignin enhances the alkaline delignification and enzymatic saccharification of cell walls

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Elumalai Sasikumar

2012-08-01

Full Text Available Abstract Background Lignin is an integral component of the plant cell wall matrix but impedes the conversion of biomass into biofuels. The plasticity of lignin biosynthesis should permit the inclusion of new compatible phenolic monomers such as flavonoids into cell wall lignins that are consequently less recalcitrant to biomass processing. In the present study, epigallocatechin gallate (EGCG was evaluated as a potential lignin bioengineering target for rendering biomass more amenable to processing for biofuel production. Results In vitro peroxidase-catalyzed polymerization experiments revealed that both gallate and pyrogallyl (B-ring moieties in EGCG underwent radical cross-coupling with monolignols mainly by β–O–4-type cross-coupling, producing benzodioxane units following rearomatization reactions. Biomimetic lignification of maize cell walls with a 3:1 molar ratio of monolignols and EGCG permitted extensive alkaline delignification of cell walls (72 to 92% that far exceeded that for lignified controls (44 to 62%. Alkali-insoluble residues from EGCG-lignified walls yielded up to 34% more glucose and total sugars following enzymatic saccharification than lignified controls. Conclusions It was found that EGCG readily copolymerized with monolignols to become integrally cross-coupled into cell wall lignins, where it greatly enhanced alkaline delignification and subsequent enzymatic saccharification. Improved delignification may be attributed to internal trapping of quinone-methide intermediates to prevent benzyl ether cross-linking of lignin to structural polysaccharides during lignification, and to the cleavage of ester intra-unit linkages within EGCG during pretreatment. Overall, our results suggest that apoplastic deposition of EGCG for incorporation into lignin would be a promising plant genetic engineering target for improving the delignification and saccharification of biomass crops.

Production of bio-fuel ethanol from distilled grain waste eluted from Chinese spirit making process.

Science.gov (United States)

Tan, Li; Sun, Zhaoyong; Zhang, Wenxue; Tang, Yueqin; Morimura, Shigeru; Kida, Kenji

2014-10-01

Distilled grain waste eluted from Chinese spirit making is rich in carbohydrates, and could potentially serve as feedstock for the production of bio-fuel ethanol. Our study evaluated two types of saccharification methods that convert distilled grain waste to monosaccharides: enzymatic saccharification and concentrated H2SO4 saccharification. Results showed that enzymatic saccharification performed unsatisfactorily because of inefficient removal of lignin during pretreatment. Concentrated H2SO4 saccharification led to a total sugar recovery efficiency of 79.0 %, and to considerably higher sugar concentrations than enzymatic saccharification. The process of ethanol production from distilled grain waste based on concentrated H2SO4 saccharification was then studied. The process mainly consisted of concentrated H2SO4 saccharification, solid-liquid separation, decoloration, sugar-acid separation, oligosaccharide hydrolysis, and continuous ethanol fermentation. An improved simulated moving bed system was employed to separate sugars from acid after concentrated H2SO4 saccharification, by which 95.8 % of glucose and 85.8 % of xylose went into the sugar-rich fraction, while 83.3 % of H2SO4 went into the acid-rich fraction. A flocculating yeast strain, Saccharomyces cerevisiae KF-7, was used for continuous ethanol fermentation, which produced an ethanol yield of 91.9-98.9 %, based on glucose concentration.

Statistical Optimization of Culture Variables for Enhancing Agarase Production by Dendryphiella arenaria Utilizing Palisada perforata (Rhodophyta) and Enzymatic Saccharification of the Macroalgal Biomass.

Science.gov (United States)

Gomaa, Mohamed; Hifney, Awatief F; Fawzy, Mustafa A; Abdel-Gawad, Khayria M

2017-12-01

Agarase is a promising biocatalyst for several industrial applications. Agarase production was evaluated by the marine fungus Dendryphiella arenaria utilizing Palisada perforata as a basal substrate in semi-solid state fermentation. Seaweed biomass, glucose, and sucrose were the most significant parameters affecting agarase production, and their levels were further optimized using Box-Behnken design. The maximum agarase activity was 7.69 U/mL. Agarase showed a degree of thermostability with half-life of 99 min at 40 °C, and declining to 44.72 min at 80 °C. Thermodynamics suggested an important process of protein aggregation during thermal inactivation. Additionally, the enzymatic saccharification of the seaweed biomass using crude agarase was optimized with respect to biomass particle size, solid/liquid ratio, and enzyme loadings. The amount of biosugars obtained after optimization was 26.15 ± 1.43 mg/g. To the best of our knowledge, this is the first report on optimization of agarase in D. arenaria.

Optimization of enzymatic saccharification of Chaetomorpha linum biomass for the production of macroalgae-based third generation bioethanol

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Ahmed Slaheddine Masmoudi

2016-08-01

Full Text Available To evaluate the efficacy of marine macro-algae Chaetomorpha linum as a potential biofuel resource, the effects of the enzymatic treatment conditions on sugar yield were evaluated using a three factor three level Box-Behnken design. The hydrothermally pretreated C. linum biomass was treated with Aspergillus niger cellulase at various liquid to solid ratios (50–100 mL/g, enzyme concentrations (10–60 U/g and incubations times (4–44 h. Data obtained from the response surface methodology were subjected to the analysis of variance and analyzed using a second order polynomial equation. The fitted model was found to be robust and was used to optimize the sugar yield (% during enzymatic hydrolysis. The optimum saccharification conditions were: L/S ratio 100 mL/g; enzyme concentration 52 U/g; and time 44 h. Their application led to a maximum sugar yield of 30.2 g/100g dry matter. Saccharomyces cerevisiae fermentation of the algal hydrolysate provided 8.6 g ethanol/100g dry matter. These results showed a promising future of applying C. linum biomass as potential feedstock for third generation bioethanol production.

Glucosidase: microbial production and effect on enzymatic hydrolysis of cellulose

Energy Technology Data Exchange (ETDEWEB)

Sternberg, D

1977-01-01

The enzymic conversion of cellulose is catalyzed by a multiple enzyme system. The Trichoderma enzyme system has insufficient ..beta..-glucosidase (EC 3.2.1.21) activity for the practical saccharification of cellulose. Aspergillus niger and A. phoenicis were superior producers of ..beta.. glucosidase and a method for production of this enzyme in liquid culture is presented. When Trichoderma cellulase preparations are supplemented with ..beta.. glucosidase from Aspergullus during practical saccharifications glucose is the predominant product and the rate of saccharification is significantly increased. The stimulatory effect of ..beta.. glucosidase appears to be due to the removal of inhibitory levels of cellobiose.

Thermo-chemical pretreatment and enzymatic hydrolysis for enhancing saccharification of catalpa sawdust.

Science.gov (United States)

Jin, Shuguang; Zhang, Guangming; Zhang, Panyue; Li, Fan; Fan, Shiyang; Li, Juan

2016-04-01

To improve the reducing sugar production from catalpa sawdust, thermo-chemical pretreatments were examined and the chemicals used including NaOH, Ca(OH)2, H2SO4, and HCl. The hemicellulose solubilization and cellulose crystallinity index (CrI) were significantly increased after thermo-alkaline pretreatments, and the thermo-Ca(OH)2 pretreatment showed the best improvement for reducing sugar production comparing to other three pretreatments. The conditions of thermo-Ca(OH)2 pretreatment and enzymatic hydrolysis were systematically optimized. Under the optimal conditions, the reducing sugar yield increased by 1185.7% comparing to the control. This study indicates that the thermo-Ca(OH)2 pretreatment is ideal for the saccharification of catalpa sawdust and that catalpa sawdust is a promising raw material for biofuel. Copyright © 2016 Elsevier Ltd. All rights reserved.

Low melting point pyridinium ionic liquid pretreatment for enhancing enzymatic saccharification of cellulosic biomass.

Science.gov (United States)

Uju; Nakamoto, Aya; Shoda, Yasuhiro; Goto, Masahiro; Tokuhara, Wataru; Noritake, Yoshiyuki; Katahira, Satoshi; Ishida, Nobuhiro; Ogino, Chiaki; Kamiya, Noriho

2013-05-01

The potential of 1-hexylpyridinium chloride ([Hpy][Cl]), to pretreat cellulosic feedstocks was investigated using microcrystalline cellulose (Avicel) and Bagasse at 80 °C or 100 °C. Short [Hpy][Cl] pretreatments, conversion of pretreated Avicel to glucose was attained after 24h enzymatic saccharification under optimal conditions, whereas regenerated Bagasse showed 1-3-fold higher conversion than untreated biomass. FT-IR analysis of both Avicel and Bagasse samples pretreated with [Hpy][Cl] or 1-ethyl-3-methyimidazolium acetate ([Emim][OAc]) revealed that these ionic liquids behaved differently during pretreatment. [Hpy][Cl] pretreatment for an extended duration (180 min) released mono- and disaccharides without using cellulase enzymes, suggesting [Hpy][Cl] has capability for direct saccharification of cellulosic feedstocks. On the basis of the results obtained, [Hpy][Cl] pretreatment enhanced initial reaction rates in enzymatic saccharification by either crystalline polymorphic alteration of cellulose or partial degradation of the crystalline cellulosic fraction in biomass. Copyright © 2012 Elsevier Ltd. All rights reserved.

Xylanase production from Thermomyces lanuginosus VAPS-24 using low cost agro-industrial residues via hybrid optimization tools and its potential use for saccharification.

Science.gov (United States)

Kumar, Vishal; Chhabra, Deepak; Shukla, Pratyoosh

2017-11-01

The xylanase production from Thermomyces lanuginosus VAPS-24 has been optimized using OFAT (One factor at a time) approach using agro-industrial substrates. Further, central composite design (CCD) has been employed to optimize various process parameters such as temperature (45-55°C), carbon source concentration (1.5-2.5%), fermentation time (72-120h) and production medium pH (6-8). Maximum xylanase yield after RSM optimization was approximately double (119.91±2.53UmL -1 ) than un-optimized conditions (61.09±0.91UmL -1 ). Several hybrid statistical tools such as Genetic Algorithm-Response Surface Methodology (GA-RSM), Artificial Neural Network (ANN), Genetic Algorithm-Artificial Neural Network (GA-ANN) were employed to obtain more optimized process parameters to maximize the xylanase production and observed an increase of 10.50% xylanase production (132.51±3.27UmL -1 ) as compared to RSM response (119.91±2.53UmL -1 ). The various pretreated and untreated agricultural residues were subjected to saccharification by using crude xylanase in which the pretreated rice straw yielded maximum fermentable sugars 126.89mgg -1 . Copyright © 2017 Elsevier Ltd. All rights reserved.

Development of an integrated approach for α-pinene recovery and sugar production from loblolly pine using ionic liquids

DEFF Research Database (Denmark)

Papa, Gabriella; Kirby, James; Murthy Konda, N. V. S. N.

2017-01-01

perspective for the production of advanced cellulosic biofuels. To date, there have been very few examples where a single conversion process has enabled recovery of both terpenes and fermentable sugars in an integrated fashion. We have used the ionic liquid (IL), 1-ethyl-3-methylimidazolium acetate [C2C1Im......][OAc] at 120 °C and 160 °C in conjunction with analytical protocols using GC-MS, to extract α-pinene and simultaneously pretreat the pine to generate high yields of fermentable sugars after saccharification. Compared to solvent extraction, the IL process enabled higher recovery rates for α-pinene, from three...... tissues type of loblolly pine, i.e. pine chips from forest residues (FC), stems from young pine (YW) and lighter wood (LW), while also generating high yields of fermentable sugars following saccharification. We propose that this combined terpene extraction/lignocellulose pretreatment approach may provide...

Biomass enzymatic saccharification is determined by the non-KOH-extractable wall polymer features that predominately affect cellulose crystallinity in corn.

Science.gov (United States)

Jia, Jun; Yu, Bin; Wu, Leiming; Wang, Hongwu; Wu, Zhiliang; Li, Ming; Huang, Pengyan; Feng, Shengqiu; Chen, Peng; Zheng, Yonglian; Peng, Liangcai

2014-01-01

Corn is a major food crop with enormous biomass residues for biofuel production. Due to cell wall recalcitrance, it becomes essential to identify the key factors of lignocellulose on biomass saccharification. In this study, we examined total 40 corn accessions that displayed a diverse cell wall composition. Correlation analysis showed that cellulose and lignin levels negatively affected biomass digestibility after NaOH pretreatments at pcorn samples indicated that cellulose and lignin should not be the major factors on biomass saccharification after pretreatments with NaOH and H2SO4 at three concentrations. Notably, despite that the non-KOH-extractable residues covered 12%-23% hemicelluloses and lignin of total biomass, their wall polymer features exhibited the predominant effects on biomass enzymatic hydrolysis including Ara substitution degree of xylan (reverse Xyl/Ara) and S/G ratio of lignin. Furthermore, the non-KOH-extractable polymer features could significantly affect lignocellulose crystallinity at pcorn.

Co-production of bio-ethanol, xylonic acid and slow-release nitrogen fertilizer from low-cost straw pulping solid residue.

Science.gov (United States)

Huang, Chen; Ragauskas, Arthur J; Wu, Xinxing; Huang, Yang; Zhou, Xuelian; He, Juan; Huang, Caoxing; Lai, Chenhuan; Li, Xin; Yong, Qiang

2018-02-01

A novel bio-refinery sequence yielding varieties of co-products was developed using straw pulping solid residue. This process utilizes neutral sulfite pretreatment which under optimal conditions (160 °C and 3% (w/v) sulfite charge) provides 64.3% delignification while retaining 90% of cellulose and 67.3% of xylan. The pretreated solids exhibited excellent enzymatic digestibility, with saccharification yields of 86.9% and 81.1% for cellulose and xylan, respectively. After pretreatment, the process of semi-simultaneous saccharification and fermentation (S-SSF) and bio-catalysis was investigated. The results revealed that decreased ethanol yields were achieved when solid loading increased from 5% to 30%. An acceptable ethanol yield of 76.8% was obtained at 20% solid loading. After fermentation, bio-catalysis of xylose remaining in fermentation broth resulted in near 100% xylonic acid (XA) yield at varied solid loadings. To complete the co-product portfolio, oxidation ammoniation of the dissolved lignin successfully transformed it into biodegradable slow-release nitrogen fertilizer with excellent agricultural properties. Copyright © 2017 Elsevier Ltd. All rights reserved.

Production by Tobacco Transplastomic Plants of Recombinant Fungal and Bacterial Cell-Wall Degrading Enzymes to Be Used for Cellulosic Biomass Saccharification.

Science.gov (United States)

Longoni, Paolo; Leelavathi, Sadhu; Doria, Enrico; Reddy, Vanga Siva; Cella, Rino

2015-01-01

Biofuels from renewable plant biomass are gaining momentum due to climate change related to atmospheric CO2 increase. However, the production cost of enzymes required for cellulosic biomass saccharification is a major limiting step in this process. Low-cost production of large amounts of recombinant enzymes by transgenic plants was proposed as an alternative to the conventional microbial based fermentation. A number of studies have shown that chloroplast-based gene expression offers several advantages over nuclear transformation due to efficient transcription and translation systems and high copy number of the transgene. In this study, we expressed in tobacco chloroplasts microbial genes encoding five cellulases and a polygalacturonase. Leaf extracts containing the recombinant enzymes showed the ability to degrade various cell-wall components under different conditions, singly and in combinations. In addition, our group also tested a previously described thermostable xylanase in combination with a cellulase and a polygalacturonase to study the cumulative effect on the depolymerization of a complex plant substrate. Our results demonstrate the feasibility of using transplastomic tobacco leaf extracts to convert cell-wall polysaccharides into reducing sugars, fulfilling a major prerequisite of large scale availability of a variety of cell-wall degrading enzymes for biofuel industry.

Effect of thermostable α-amylase injection on mechanical and physiochemical properties for saccharification of extruded corn starch.

Science.gov (United States)

Myat, Lin; Ryu, Gi-Hyung

2014-01-30

In industry, a jet cooker is used to gelatinize starch by mixing the starch slurry with steam under pressure at 100-175 °C. A higher degree of starch hydrolysis in an extruder is possible with glucoamylase. Unfortunately, it is difficult to carry out liquefaction and saccharification in parallel, because the temperature of gelatinization will be too high and will inactivate glucoamylase. Since the temperature for liquefaction and saccharification is different, it is hard to change the temperature from high (required for liquefaction) to low (required for saccharification). The industrial gelatinization process is usually carried out with 30-35% (w/w) dry solids starch slurry. Conventional jet cookers cannot be used any more at high substrate concentrations owing to higher viscosity. In this study, therefore, corn starch was extruded at different melt temperatures to overcome these limitations and to produce the highest enzyme-accessible starch extrudates. Significant effects on physical properties (water solubility index, water absorption index and color) and chemical properties (reducing sugar and % increase in reducing sugar after saccharification) were achieved by addition of thermostable α-amylase at melt temperatures of 115 and 135 °C. However, there was no significant effect on % increase in reducing sugar of extruded corn starch at 95 °C. The results show the great potential of extrusion with thermostable α-amylase injection at 115 and 135 °C as an effective pretreatment for breaking down starch granules, because of the significant increase (P < 0.05) in % reducing sugar and enzyme-accessible extrudates for saccharification yield. © 2013 Society of Chemical Industry.

Direct saccharification of milo

Energy Technology Data Exchange (ETDEWEB)

1981-10-28

Crude or unrefined milo grain is pulverized to less than 100 mesh, suspended in water, and directly saccharified with enzymes. Thus, 10 kg of pulverized milo (100 mesh) was suspended in 20 liters water, liquified in the presence of bacterial amylase at 90/sup 0/ for 30 minutes and autoclaved for 10 minutes at 120/sup 0/, the liquifaction autoclave procedures were then repeated. The liquified preparation was saccharified in the presence of bacterial glucoamylase at 55/sup 0/ for 72 hours. The percentage saccharification of unrefined milo was 92-97%, which was comparable to that obtained with refined milo or a milo starch preparation. The saccharified milo preparation, when used as a sugar source in glutamic acid, lysine, or inosine fermentation, also showed similar efficiency to that of saccharified milo starch.

Technology Evaluation of Process Configurations for Second Generation Bioethanol Production using Dynamic Model-based Simulations

DEFF Research Database (Denmark)

Morales Rodriguez, Ricardo; Meyer, Anne S.; Gernaey, Krist

2011-01-01

An assessment of a number of different process flowsheets for bioethanol production was performed using dynamic model-based simulations. The evaluation employed diverse operational scenarios such as, fed-batch, continuous and continuous with recycle configurations. Each configuration was evaluated...... against the following benchmark criteria, yield (kg ethanol/kg dry-biomass), final product concentration and number of unit operations required in the different process configurations. The results has shown the process configuration for simultaneous saccharification and co-fermentation (SSCF) operating...... in continuous mode with a recycle of the SSCF reactor effluent, results in the best productivity of bioethanol among the proposed process configurations, with a yield of 0.18 kg ethanol /kg dry-biomass....

Enzymatic saccharification of seaweeds into fermentable sugars by xylanase from marine Bacillus sp. strain BT21.

Science.gov (United States)

Parab, Pankaj; Khandeparker, Rakhee; Amberkar, Ujwala; Khodse, Vishwas

2017-10-01

Enzymatic hydrolysis of seaweed biomass was studied using xylanase produced from marine bacteria Bacillus sp. strain BT21 through solid-state fermentation of wheat bran. Three types of seaweeds, Ahnfeltia plicata , Padina tetrastromatica and Ulva lactuca , were selected as representatives of red, brown, and green seaweeds, respectively. Seaweed biomass was pretreated with hot water. The efficiency of pretreated biomass to release reducing sugar by the action of xylanase as well as the type of monosaccharide released during enzyme saccharification of seaweed biomass was studied. It was seen that pretreated biomass of seaweed A. plicata, U. lactuca , and P. tetrastroma , at 121 °C for 45 min, followed by incubation with 50 IU xylanase released reducing sugars of 233 ± 5.3, 100 ± 6.1 and 73.3 ± 4.1 µg/mg of seaweed biomass, respectively. Gas chromatography analysis illustrated the release of xylose, glucose, and mannose during the treatment process. Hot water pre-treatment process enhanced enzymatic conversion of biomass into sugars. This study revealed the important role of xylanase in saccharification of seaweed, a promising feedstock for third-generation bioethanol production.

Optimization of endoglucanase production from thermophilic strain of Bacillus licheniformis RT-17 and its application for saccharification of sugarcane bagasse

International Nuclear Information System (INIS)

Tariq, R.; Qadir, F.; Ahmed, A.; Shariq, M.; Zafar, U.; Khan, S.A.

2018-01-01

Thermostable cellulases are required for a variety of commercial processes. Bacillus is a house of thermostable proteins. Screening of indigenously isolated strains of bacteria revealed the promising production of cellulase by a strain, RT-17, at 50 degree C. The strain was identified on the basis of biochemical and molecular characteristics as B. licheniformis. The factors affecting cellulase production from B. licheniformis RT-17 were evaluated for their significant effect using Plackett Burman Design and were optimized by employing Box-Behnken Design. The model predicted 9.808 IU/ml of endoglucanase (EG) under optimum conditions of 50 degree C; 10% inoculum size; pH 5; and 1% peptone in fermentation medium. Practically, a titer of 9.128 IU/ml was obtained, showed the validity of the model. The enzyme preparation from B. licheniformis RT-17 was applied in combination with xylanase and pectinase preparations from indigenous yeasts for the hydrolysis of sugarcane bagasse (SCB). A higher degree of synergy (7.1 folds) was observed when yeast pectinase was used with bacterial cellulase for the hydrolysis of alkali treated SCB. Whereas, the degree of synergy was lower when bacterial cellulase was mixed with yeast xylanase. The study revealed the possibility of utilization of combination of yeast and bacterial enzymes for biomass saccharification. (author)

Modelling the bioconversion of cellulose into microbial products: rate limitations

Energy Technology Data Exchange (ETDEWEB)

Asenjo, J A

1984-12-01

The direct bioconversion of cellulose into microbial products carried out as a simultaneous saccharification and fermentation has a strong effect on the rates of cellulose degradation because cellobiose and glucose inhibition of the reaction are circumvented. A general mathematical model of the kinetics of this bioconversion has been developed. Its use in representing aerobic systems and in the analysis of the kinetic limitations has been investigated. Simulations have been carried out to find the rate limiting steps in slow fermentations and in rapid ones as determined by the specific rate of product formation. The requirements for solubilising and depolymerising enzyme activities (cellulase and cellobiase) in these systems has been determined. The activity that have been obtained for fungal cellulases are adequate for the kinetic requirements of the fastest fermentative strains. The results also show that for simultaneous bioconversions where strong cellobiose and glucose inhibition is overcome, no additional cellobiase is necessary to increase the rate of product formation. These results are useful for the selection of cellolytic micro-organisms and in the determination of enzymes to be cloned in recombinant strains. 17 references.

Final Technical Report - Consolidating Biomass Pretreatment with Saccharification by Resolving the Spatial Control Mechanisms of Fungi

Energy Technology Data Exchange (ETDEWEB)

Schilling, Jonathan [Univ. of Minnesota, Minneapolis, MN (United States)

2017-07-06

Consolidated bioprocessing (CBP) of lignocellulose combines enzymatic sugar release (saccharification) with fermentation, but pretreatments remain separate and costly. In nature, lignocellulose-degrading brown rot fungi consolidate pretreatment and saccharification, likely using spatial gradients to partition these incompatible reactions. With the field of biocatalysis maturing, reaction partitioning is increasingly reproducible for commercial use. Therefore, my goal was to resolve the reaction partitioning mechanisms of brown rot fungi so that they can be applied to bioconversion of lignocellulosic feedstocks. Brown rot fungi consolidate oxidative pretreatments with saccharification and are a focus for biomass refining because 1) they attain >99% sugar yield without destroying lignin, 2) they use a simplified cellulase suite that lacks exoglucanase, and 3) their non-enzymatic pretreatment is facilitative and may be accelerated. Specifically, I hypothesized that during brown rot, oxidative pretreatments occur ahead of enzymatic saccharification, spatially, and the fungus partitions these reactions using gradients in pH, lignin reactivity, and plant cell wall porosity. In fact, we found three key results during these experiments for this work: 1) Brown rot fungi have an inducible cellulase system, unlike previous descriptions of a constitutive mechanism. 2) The induction of cellulases is delayed until there is repression of oxidatively-linked genes, allowing the brown rot fungi to coordinate two incompatible reactions (oxidative pretreatment with enzymatic saccharification, to release wood sugars) in the same pieces of wood. 3) This transition is mediated by the same wood sugar, cellobiose, released by the oxidative pretreatment step. Collectively, these findings have been published in excellent journal outlets and have been presented at conferences around the United States, and they offer clear targets for gene discovery en route to making biofuels and biochemicals

Cold alkaline extraction as a pretreatment for bioethanol production from eucalyptus, sugarcane bagasse and sugarcane straw

International Nuclear Information System (INIS)

Carvalho, Danila Morais de; Sevastyanova, Olena; Queiroz, José Humberto de; Colodette, Jorge Luiz

2016-01-01

Highlights: • Mathematical approach to optimize the process of cold alkaline extraction. • Hemicelluloses and lignin removal from biomasses by cold alkaline extraction. • Higher xylan and lignin removal for straw during pretreatment. • Formation of pseudo-extractives for eucalyptus during pretreatment. • Higher ethanol production for pretreated sugarcane straw. - Abstract: Optimal conditions for the cold alkaline extraction (CAE) pretreatment of eucalyptus, sugarcane bagasse and sugarcane straw are proposed in view of their subsequent bioconversion into ethanol through the semi-simultaneous saccharification and fermentation (SSSF) process (with presaccharification followed by simultaneous saccharification and fermentation, or SSF). The optimum conditions, which are identified based on an experiment with a factorial central composite design, resulted in the removal of 46%, 52% and 61% of the xylan and 15%, 37% and 45% of the lignin for eucalyptus, bagasse and straw, respectively. The formation of pseudo-extractives was observed during the CAE of eucalyptus. Despite the similar glucose concentration and yield for all biomasses after 12 h of presaccharification, the highest yield (0.065 g_e_t_h_a_n_o_l/g_b_i_o_m_a_s_s), concentrations (5.74 g L"−"1) and volumetric productivity for ethanol (0.57 g L"−"1 h"−"1) were observed for the sugarcane straw. This finding was most likely related to the improved accessibility of cellulose that resulted from the removal of the largest amount of xylan and lignin.

Biomass Enzymatic Saccharification Is Determined by the Non-KOH-Extractable Wall Polymer Features That Predominately Affect Cellulose Crystallinity in Corn

OpenAIRE

Jia, Jun; Yu, Bin; Wu, Leiming; Wang, Hongwu; Wu, Zhiliang; Li, Ming; Huang, Pengyan; Feng, Shengqiu; Chen, Peng; Zheng, Yonglian; Peng, Liangcai

2014-01-01

Corn is a major food crop with enormous biomass residues for biofuel production. Due to cell wall recalcitrance, it becomes essential to identify the key factors of lignocellulose on biomass saccharification. In this study, we examined total 40 corn accessions that displayed a diverse cell wall composition. Correlation analysis showed that cellulose and lignin levels negatively affected biomass digestibility after NaOH pretreatments at p

Enzymatic saccharification and structural properties of industrial wood sawdust: Recycled ionic liquids pretreatments

International Nuclear Information System (INIS)

Auxenfans, Thomas; Buchoux, Sébastien; Larcher, Dominique; Husson, Gérard; Husson, Eric; Sarazin, Catherine

2014-01-01

Highlights: • 1-Ethyl-3-metylimidazolium acetate is an effective catalyst for pretreatment of hardwood and softwood sawdust. • Regeneration of cellulosic fraction from ionic liquid is discussed. • 1-Ethyl-3-methylimidazolium acetate can be reused at least 7 times without loss of its efficiency. • Removal of extractives and lignin with slight cellulose and xylan losses were observed. • Better cellulase accessibility to cellulose thanks to the expansion of the powder and the creation of a large porous volume. - Abstract: Wood residues constitute a promising challenge for biochemical processing into bioethanol and chemicals with competitive costs. Here, we report the impacts of pretreatments in a hydrophilic ionic liquid ([C2mim][OAc]), onto the physicochemical properties and enzymatic saccharification of softwood (spruce) and hardwood (oak) sawdust. Enzymatic saccharification of IL- pretreated sawdust is significantly increased (up to 7 times) when compared to untreated ones. Methanol, ethanol or water can be used as polar anti-solvent for the recovery of a cellulose rich fraction after dissolution in IL (i.e regeneration step) without any effect on enzymatic saccharification. Chemical, textural and structural modifications possibly induced by the IL pretreatments have been investigated through various means (Infra-red spectroscopy, NMR, X-ray diffraction) in order to correlate the observed modifications in enzymatic saccharification. This mild pretreatment seemed to mainly act in a breakdown of lignocellulosic organization leading to better cellulase accessibility to cellulose thanks to the expansion of the powder and the creation of a large porous volume (5 times more apparent porous volume). Partial removal of lignin and extractives may also contribute to the best enzymatic performances. The recyclability and reuse up to 7 times of [C2mim][OAc] is shown without the need of strictly anhydrous conditions and any alteration of the pretreatment

Biomass Enzymatic Saccharification Is Determined by the Non-KOH-Extractable Wall Polymer Features That Predominately Affect Cellulose Crystallinity in Corn

Science.gov (United States)

Wu, Leiming; Wang, Hongwu; Wu, Zhiliang; Li, Ming; Huang, Pengyan; Feng, Shengqiu; Chen, Peng; Zheng, Yonglian; Peng, Liangcai

2014-01-01

Corn is a major food crop with enormous biomass residues for biofuel production. Due to cell wall recalcitrance, it becomes essential to identify the key factors of lignocellulose on biomass saccharification. In this study, we examined total 40 corn accessions that displayed a diverse cell wall composition. Correlation analysis showed that cellulose and lignin levels negatively affected biomass digestibility after NaOH pretreatments at pbiomass saccharification after pretreatments with NaOH and H2SO4 at three concentrations. Notably, despite that the non-KOH-extractable residues covered 12%–23% hemicelluloses and lignin of total biomass, their wall polymer features exhibited the predominant effects on biomass enzymatic hydrolysis including Ara substitution degree of xylan (reverse Xyl/Ara) and S/G ratio of lignin. Furthermore, the non-KOH-extractable polymer features could significantly affect lignocellulose crystallinity at pbiomass digestibility. Hence, this study could suggest an optimal approach for genetic modification of plant cell walls in bioenergy corn. PMID:25251456

Studies on the conversion of cellulose hydrolysate into citric acid by Aspergillus niger

Energy Technology Data Exchange (ETDEWEB)

Manonmani, H.K.; Sreekantiah, K.R.

1987-06-01

The production of citric acid by Aspergillus niger (16) was studied using enzymatic hydrolysate of alkali-treated bagasse by solid state fermentation. Saccharification and fermentations were carried out sequentially as well as simultaneously. Conditions for optimum citric acid production using cellulose hydrolysate medium were: sugar concentration: 7% (w/w); NaNO/sub 3/; 400 mg/N/sub 2//l medium; KH/sub 2/PO/sub 4/:/0.1%/l medium; ethanol: 3% (v/w); 1 ml of 1 x 10 squared m fluoroacetate and coconut oil: 3% (v/w). Simultaneous saccharification and fermentation was not found to be suitable for citric acid production. 44% conversion of total reducing sugars to citric acid was obtained in 72 hours fermentation by sequential process with the above mentioned parameters. (Refs. 15).

Sequential and simultaneous strategies for biorefining of wheat straw using room temperature ionic liquids, xylanases and cellulases.

Science.gov (United States)

Husson, Eric; Auxenfans, Thomas; Herbaut, Mickael; Baralle, Manon; Lambertyn, Virginie; Rakotoarivonina, Harivoni; Rémond, Caroline; Sarazin, Catherine

2018-03-01

Sequential and simultaneous strategies for fractioning wheat straw were developed in combining 1-ethyl-3-methyl imidazolium acetate [C2mim][OAc], endo-xylanases from Thermobacillus xylanilyticus and commercial cellulases. After [C2mim][OAc]-pretreatment, hydrolysis catalyzed by endo-xylanases of wheat straw led to efficient xylose production with very competitive yield (97.6 ± 1.3%). Subsequent enzymatic saccharification allowed achieving a total degradation of cellulosic fraction (>99%). These high performances revealed an interesting complementarity of [C2mim][OAc]- and xylanase-pretreatments for increasing enzymatic digestibility of cellulosic fraction in agreement with the structural and morphological changes of wheat straw induced by each of these pretreatment steps. In addition a higher tolerance of endo-xylanases from T. xylaniliticus to [C2mim][AcO] until 30% v/v than cellulases from T. reesei was observed. Based on this property, a simultaneous strategy combining [C2mim][OAc]- and endo-xylanases as pretreatment in a one-batch produced xylose with similar yield than those obtained by the sequential strategy. Copyright © 2017 Elsevier Ltd. All rights reserved.

Development of cellulase-nanoconjugates with enhanced ionic liquid and thermal stability for in situ lignocellulose saccharification.

Science.gov (United States)

Grewal, Jasneet; Ahmad, Razi; Khare, S K

2017-10-01

The present work aimed to improve catalytic efficiency of Trichoderma reesei cellulase for enhanced saccharification. The cellulase was immobilized on two nanomatrices i.e. magnetic and silica nanoparticles with immobilization efficiency of 85% and 76% respectively. The nanobioconjugates exhibited increase in V max , temperature optimum, pH and thermal stability as compared with free enzyme. These could be efficiently reused for five repeated cycles and were stable in 1-ethyl-3-methylimidazoliumacetate [EMIM][Ac], an ionic liquid. Ionic liquids (IL) are used as green solvents to dissolve lignocellulosic biomass and facilitate better saccharification. The cellulase immobilized on magnetic nanoparticles was used for in situ saccharification of [EMIM][Ac] pretreated sugarcane bagasse and wheat straw for two cycles. The structural deconstruction and decrease in biomass crystallinity was confirmed by SEM, XRD and FTIR. The high hydrolysis yields (∼89%) obtained in this one-pot process coupled with IL stability and recycled use of immobilized cellulase, potentiates its usefulness in biorefineries. Copyright © 2017 Elsevier Ltd. All rights reserved.

Biological utilization of bagasse, a lignocellulose waste

CSIR Research Space (South Africa)

Paterson-Jones, JC

1989-01-01

Full Text Available for the production of single cell protein from the hemicelluloses and cellulose hydrolysates and the production of ethanol from the the cellulose by simultaneous saccharification and fermentation and from the hemicelluloses hydroly-sate by direct fermentation...

Production by Tobacco Transplastomic Plants of Recombinant Fungal and Bacterial Cell-Wall Degrading Enzymes to Be Used for Cellulosic Biomass Saccharification

Directory of Open Access Journals (Sweden)

Paolo Longoni

2015-01-01

Full Text Available Biofuels from renewable plant biomass are gaining momentum due to climate change related to atmospheric CO2 increase. However, the production cost of enzymes required for cellulosic biomass saccharification is a major limiting step in this process. Low-cost production of large amounts of recombinant enzymes by transgenic plants was proposed as an alternative to the conventional microbial based fermentation. A number of studies have shown that chloroplast-based gene expression offers several advantages over nuclear transformation due to efficient transcription and translation systems and high copy number of the transgene. In this study, we expressed in tobacco chloroplasts microbial genes encoding five cellulases and a polygalacturonase. Leaf extracts containing the recombinant enzymes showed the ability to degrade various cell-wall components under different conditions, singly and in combinations. In addition, our group also tested a previously described thermostable xylanase in combination with a cellulase and a polygalacturonase to study the cumulative effect on the depolymerization of a complex plant substrate. Our results demonstrate the feasibility of using transplastomic tobacco leaf extracts to convert cell-wall polysaccharides into reducing sugars, fulfilling a major prerequisite of large scale availability of a variety of cell-wall degrading enzymes for biofuel industry.

A novel method to determine simultaneously methane production during in vitro gas production using fully automated equipment

NARCIS (Netherlands)

Pellikaan, W.F.; Hendriks, W.H.; Uwimanaa, G.; Bongers, L.J.G.M.; Becker, P.M.; Cone, J.W.

2011-01-01

An adaptation of fully automated gas production equipment was tested for its ability to simultaneously measure methane and total gas. The simultaneous measurement of gas production and gas composition was not possible using fully automated equipment, as the bottles should be kept closed during the

Enhanced Ethanol Production with Mixed Lignocellulosic Substrates from Commercial Furfural and Cassava Residues

Directory of Open Access Journals (Sweden)

Li Ji

2015-01-01

Full Text Available Simultaneous saccharification and fermentation (SSF is an attractive process configuration for bio-ethanol production. Further reductions in process cost of SSF are expected with the use of waste agricultural or industrial materials as feedstock. In the current study, two industrial lignocellulosic wastes, cassava residues (CR and furfural residues (FR, were combined during SSF for ethanol production due to their value-added applications and positive environmental impacts. After CR were liquefied and saccharified, saccharification liquid was added to SSF of FR. The effect of substrate fractions was investigated in terms of ethanol yield, byproduct concentration and the number of yeast cells. Besides, a natural surfactant, Gleditsia saponin, was added to investigate the effect of FR lignin on SSF with 20% substrate concentration. The results showed that increasing the ratio of CR/FR improved the ethanol yield and that the ethanol yield was also increased gradually by increasing the substrate concentration from 6% to 12%. A high ethanol concentration of 36.0 g/L was obtained under the condition of CR:FR = 2:1 with 12% substrate concentration, reaching 71.1% of the theoretical yield. However, Gleditsia saponin did not affect the ethanol yield, indicating the insignificant effect of lignin in SSF with low lignin content in the reaction system.

An integrated bioconversion process for the production of L-lactic acid from starchy feedstocks

Energy Technology Data Exchange (ETDEWEB)

Tsai, S.P.; Moon, S.H.

1997-07-01

The potential market for lactic acid as the feedstock for biodegradable polymers, oxygenated chemicals, and specialty chemicals is significant. L-lactic acid is often the desired enantiomer for such applications. However, stereospecific lactobacilli do not metabolize starch efficiently. In this work, Argonne researchers have developed a process to convert starchy feedstocks into L-lactic acid. The processing steps include starch recovery, continuous liquefaction, and simultaneous saccharification and fermentation. Over 100 g/L of lactic acid was produced in less than 48 h. The optical purity of the product was greater than 95%. This process has potential economical advantages over the conventional process.

DIFFERENCES BETWEEN WHEAT CULTIVARS IN GRAIN PARAMETERS RELATED TO ETHANOL PRODUCTION

Directory of Open Access Journals (Sweden)

Daniela Mikulíková

2011-12-01

Full Text Available Wheat grain samples from sixteen winter cultivars originated from four localities were evaluated and compared in traits related to ethanol production as grain yield, grain hardness, content of protein, starch and amylose, and α-amylase activity. Results obtained indicate significant differences between cultivars in amylose content, α-amylase activity, and grain hardness compared to grain yield, protein content, and starch content where differences were not significant. The amylose content, α-amylase activity, and grain hardness were affected by cultivar. Both testing methods for starch fermentation - separated hydrolysis and fermentation (SHF and simultaneous saccharification and fermentation (SSF revealed difference between cultivars in ethanol yield.

LPMOs in cellulase mixtures affect fermentation strategies for lactic acid production from lignocellulosic biomass.

Science.gov (United States)

Müller, Gerdt; Kalyani, Dayanand Chandrahas; Horn, Svein Jarle

2017-03-01

Enzymatic catalysis plays a key role in the conversion of lignocellulosic biomass to fuels and chemicals such as lactic acid. In the last decade, the efficiency of commercial cellulase cocktails has increased significantly, in part due to the inclusion of lytic polysaccharide monooxygenases (LPMOs). However, the LPMOs' need for molecular oxygen to break down cellulose demands reinvestigations of process conditions. In this study, we evaluate the efficiency of lactic acid production from steam-exploded birch using an LPMO-containing cellulase cocktail in combination with lactic acid bacteria, investigating both separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF). While the SSF set up generally has been considered to be more efficient because it avoids sugar accumulation which may inhibit the cellulases, the SHF set up in our study yielded 26-32% more lactic acid than the SSF. This was mainly due to competition for oxygen between LPMOs and the fermenting organisms in the SSF process, which resulted in reduced LPMO activity and thus less efficient saccharification of the lignocellulosic substrate. By means of aeration it was possible to activate the LPMOs in the SSF, but less lactic acid was produced due to a shift in metabolic pathways toward production of acetic acid. Overall, this study shows that lactic acid can be produced efficiently from lignocellulosic biomass, but that the use of LPMO-containing cellulase cocktails in fermentation processes demands re-thinking of traditional process set ups due to the requirement of oxygen in the saccharification step. Biotechnol. Bioeng. 2017;114: 552-559. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Enzymatic saccharification of hemicellulose obtained from hydrothermally pretreated sugar can bagasse and beech bark

Energy Technology Data Exchange (ETDEWEB)

Walch, E; Zemann, A; Bonn, G; Bobleter, O [Innsbruck Univ. (Austria). Inst. fuer Radiochemie und Angewandte Physikalische Chemie; Schinner, F [Innsbruck Univ. (AT). Inst. for Microbiology

1992-01-01

Characteristics of different xylanses and their use in the saccharification of sugar cane bagasse and beech bark were studied. Bagasse was pretreated by two different hydrothermolysis procedures, a recirculation and a direct flow-through process. The recirculation procedure resulted in a higher yield of dry matter in the hydrothermolysis solution and a higher saccharification effect after enzymatic hydrolysis. In the case of beech bark, the tannins have first to be removed at temperatures of 120-140{sup o}C. In a second hydrothermal step (at approximately 200{sup o}C), a hemicellulose solution is obtained which can be saccharified enzymatically with high yields. The inhibitory effect of the tannins is experimentally demonstrated. (author).

Ethanol production from corn cobs by co-culture of Saccharomyces ...

African Journals Online (AJOL)

Saccharomyces cerevisiae and Aspergillus niger were used in a co-culture for the simultaneous saccharification and fermentation (SSF) of 1% and 10% (w/v) dry pre-treated corn cobs to ethanol. Positive controls of glucose of same concentrations in a synthetic medium were also fermented. At 1% substrate concentration, ...

Biomass saccharification is largely enhanced by altering wall polymer features and reducing silicon accumulation in rice cultivars harvested from nitrogen fertilizer supply.

Science.gov (United States)

Zahoor; Sun, Dan; Li, Ying; Wang, Jing; Tu, Yuanyuan; Wang, Yanting; Hu, Zhen; Zhou, Shiguang; Wang, Lingqiang; Xie, Guosheng; Huang, Jianliang; Alam, Aftab; Peng, Liangcai

2017-11-01

In this study, two rice cultivars were collected from experimental fields with seven nitrogen fertilizer treatments. All biomass samples contained significantly increased cellulose contents and reduced silica levels, with variable amounts of hemicellulose and lignin from different nitrogen treatments. Under chemical (NaOH, CaO, H 2 SO 4 ) and physical (hot water) pretreatments, biomass samples exhibited much enhanced hexoses yields from enzymatic hydrolysis, with high bioethanol production from yeast fermentation. Notably, both degree of polymerization (DP) of cellulose and xylose/arabinose (Xyl/Ara) ratio of hemicellulose were reduced in biomass residues, whereas other wall polymer features (cellulose crystallinity and monolignol proportion) were variable. Integrative analysis indicated that cellulose DP, hemicellulosic Xyl/Ara and silica are the major factors that significantly affect cellulose crystallinity and biomass saccharification. Hence, this study has demonstrated that nitrogen fertilizer supply could largely enhance biomass saccharification in rice cultivars, mainly by reducing cellulose DP, hemicellulosic Xyl/Ara and silica in cell walls. Copyright © 2017 Elsevier Ltd. All rights reserved.

Comparing cell viability and ethanol fermentation of the thermotolerant yeast Kluyveromyces marxianus and Saccharomyces cerevisiae on steam-exploded biomass treated with laccase.

Science.gov (United States)

Moreno, Antonio D; Ibarra, David; Ballesteros, Ignacio; González, Alberto; Ballesteros, Mercedes

2013-05-01

In this study, the thermotolerant yeast Kluyveromyces marxianus CECT 10875 was compared to the industrial strain Saccharomyces cerevisiae Ethanol Red for lignocellulosic ethanol production. For it, whole slurry from steam-exploded wheat straw was used as raw material, and two process configurations, simultaneous saccharification and fermentation (SSF) and presaccharification and simultaneous saccharification and fermentation (PSSF), were evaluated. Compared to S. cerevisiae, which was able to produce ethanol in both process configurations, K. marxianus was inhibited, and neither growth nor ethanol production occurred during the processes. However, laccase treatment of the whole slurry removed specifically lignin phenols from the overall inhibitory compounds present in the slurry and triggered the fermentation by K. marxianus, attaining final ethanol concentrations and yields comparable to those obtained by S. cerevisiae. Copyright © 2012 Elsevier Ltd. All rights reserved.

An Electrochemical Impedance Spectroscopy System for Monitoring Pineapple Waste Saccharification

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Claudia Conesa

2016-02-01

Full Text Available Electrochemical impedance spectroscopy (EIS has been used for monitoring the enzymatic pineapple waste hydrolysis process. The system employed consists of a device called Advanced Voltammetry, Impedance Spectroscopy & Potentiometry Analyzer (AVISPA equipped with a specific software application and a stainless steel double needle electrode. EIS measurements were conducted at different saccharification time intervals: 0, 0.75, 1.5, 6, 12 and 24 h. Partial least squares (PLS were used to model the relationship between the EIS measurements and the sugar determination by HPAEC-PAD. On the other hand, artificial neural networks: (multilayer feed forward architecture with quick propagation training algorithm and logistic-type transfer functions gave the best results as predictive models for glucose, fructose, sucrose and total sugars. Coefficients of determination (R2 and root mean square errors of prediction (RMSEP were determined as R2 > 0.944 and RMSEP < 1.782 for PLS and R2 > 0.973 and RMSEP < 0.486 for artificial neural networks (ANNs, respectively. Therefore, a combination of both an EIS-based technique and ANN models is suggested as a promising alternative to the traditional laboratory techniques for monitoring the pineapple waste saccharification step.

Comparison between solid-state and powder-state alkali pretreatment on saccharification and fermentation for bioethanol production from rice straw.

Science.gov (United States)

Yeasmin, Shabina; Kim, Chul-Hwan; Islam, Shah Md Asraful; Lee, Ji-Young

2016-01-01

The efficacy of different concentrations of NaOH (0.25%, 0.50%, 0.75%, and 1.00%) for the pretreatment of rice straw in solid and powder state in enzymatic saccharification and fermentation for the production of bioethanol was evaluated. A greater amount of biomass was recovered through solid-state pretreatment (3.74 g) from 5 g of rice straw. The highest increase in the volume of rice straw powder as a result of swelling was observed with 1.00% NaOH pretreatment (48.07%), which was statistically identical to 0.75% NaOH pretreatment (32.31%). The surface of rice straw was disrupted by the 0.75% NaOH and 1.00% NaOH pretreated samples as observed using field-emission scanning electron microscopy (FE-SEM) and atomic force microscopy (AFM). In Fourier-transform infrared (FT-IR) spectra, absorbance of hydroxyl groups at 1,050 cm(-1) due to the OH group of lignin was gradually decreased with the increase of NaOH concentration. The greatest amounts of glucose and ethanol were obtained in 1.00% NaOH solid-state pretreated and powder-state hydrolyzed samples (0.804 g g(-1) and 0.379 g g(-1), respectively), which was statistically similar to the use of 0.75% NaOH (0.763 g g(-1) and 0.358 g g(-1), respectively). Thus, solid-state pretreatment with 0.75% NaOH and powder-state hydrolysis appear to be suitable for fermentation and bioethanol production from rice straw.

Understanding the effects of lignosulfonate on enzymatic saccharification of pure cellulose

Science.gov (United States)

Hongming Lou; Haifeng Zhou; Xiuli Li; Mengxia Wang; J.Y. Zhu; Xueqing Qiu

2014-01-01

The effects of lignosulfonate (LS) on enzymatic saccharification of pure cellulose were studied. Four fractions of LS with different molecular weight (MW) prepared by ultrafiltration of a commercial LS were applied at different loadings to enzymatic hydrolysis of Whatman paper under different pH. Using LS fractions with low MW and high degree of sulfonation can enhance...

Utilization of Bagasse Cellulose for Ethanol Production through Simultaneous Saccharification and Fermentation by Xylanase

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M Samsuri

2010-10-01

Full Text Available Bagasse is a solid residue from sugar cane process, which is not many use it for some product which have more added value. Bagasse, which is a lignosellulosic material, be able to be use for alternative energy resources like bioethanol or biogas. With renewable energy resources a crisis of energy in Republic of Indonesia could be solved, especially in oil and gas. This research has done the conversion of bagasse to bioethanol with xylanase enzyme. The result show that bagasse contains of 52,7% cellulose, 20% hemicelluloses, and 24,2% lignin. Xylanase enzyme and Saccharomyces cerevisiae was used to hydrolyse and fermentation in SSF process. Variation in this research use pH (4, 4,5, and 5, for increasing ethanol quantity, SSF process was done by added chloride acid (HCl with concentration 0.5% and 1% (v/v and also pre-treatment with white rot fungi such as Lentinus edodes (L.edodes as long 4 weeks. The SSF process was done with 24, 48, 72, and 96 hour's incubation time for fermentation. Variation of pH 4, 4,5, and 5 can produce ethanol with concentrations 2,357 g/L, 2,451 g/L, 2,709 g/L. The added chloride acid (HCl with concentration 0.5% and 1% (v/v and L. edodes can increase ethanol yield, The highest ethanol concentration with added chloride acid (HCl concentration 0.5% and 1% consecutively is 2,967 g/L, 3,249 g/L. The highest ethanol concentration with pre-treatment by L. edodes is 3,202 g/L.

Metal accumulation by sunflower (Helianthus annuus L. and the efficacy of its biomass in enzymatic saccharification.

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Saurabh Sudha Dhiman

Full Text Available Accumulation of metal contaminants in soil as a result of various industrial and anthropogenic activities has reduced soil fertility significantly. Phytoextraction of metal contaminants can improve soil fertility and provide inexpensive feedstock for biorefineries. We investigated the hyperaccumulation capacity of sunflower (Helianthus annuus biomass by cultivating these plants in various concentrations of metal contaminants. Sunflowers were grown in soils contaminated with various levels of heavy metals (10-2,000 mg/kg dry soil. The degree of metal uptake by different parts of the biomass and the residual concentration in the soil were estimated through inductively coupled plasma mass spectrometry. An almost 2.5-fold hyperaccumulation of Zn2+ was observed in the leaf and flower biomass compared with the concentration in the soil. For the subsequent saccharification of biomass with hyperaccumulated contaminants, a fungal lignocellulosic consortium was used. The fungal consortium cocktail retained more than 95% filter paper activity with 100 mM Ni2+ ions even after 36 h. The highest saccharification yield (SY, 87.4% was observed with Ni2+ as the contaminant (10 mg/kg dry wt, whereas Pb2+ (251.9 mg/kg dry wt was the strongest inhibitor of biomass hydrolysis, resulting in only a 30% SY. Importantly, the enzyme cocktail produced by the fungal consortium resulted in almost the same SY (% as that obtained from a combination of commercial cellulase and β-glucosidase. Significant sugar conversion (61.7% from H. annuus biomass hydrolysate occurred, resulting in the production of 11.4 g/L of bioethanol. This is the first study to assess the suitability of phytoremediated sunflower biomass for bioethanol production.

Metal accumulation by sunflower (Helianthus annuus L.) and the efficacy of its biomass in enzymatic saccharification.

Science.gov (United States)

Dhiman, Saurabh Sudha; Zhao, Xin; Li, Jinglin; Kim, Dongwook; Kalia, Vipin C; Kim, In-Won; Kim, Jae Young; Lee, Jung-Kul

2017-01-01

Accumulation of metal contaminants in soil as a result of various industrial and anthropogenic activities has reduced soil fertility significantly. Phytoextraction of metal contaminants can improve soil fertility and provide inexpensive feedstock for biorefineries. We investigated the hyperaccumulation capacity of sunflower (Helianthus annuus) biomass by cultivating these plants in various concentrations of metal contaminants. Sunflowers were grown in soils contaminated with various levels of heavy metals (10-2,000 mg/kg dry soil). The degree of metal uptake by different parts of the biomass and the residual concentration in the soil were estimated through inductively coupled plasma mass spectrometry. An almost 2.5-fold hyperaccumulation of Zn2+ was observed in the leaf and flower biomass compared with the concentration in the soil. For the subsequent saccharification of biomass with hyperaccumulated contaminants, a fungal lignocellulosic consortium was used. The fungal consortium cocktail retained more than 95% filter paper activity with 100 mM Ni2+ ions even after 36 h. The highest saccharification yield (SY, 87.4%) was observed with Ni2+ as the contaminant (10 mg/kg dry wt), whereas Pb2+ (251.9 mg/kg dry wt) was the strongest inhibitor of biomass hydrolysis, resulting in only a 30% SY. Importantly, the enzyme cocktail produced by the fungal consortium resulted in almost the same SY (%) as that obtained from a combination of commercial cellulase and β-glucosidase. Significant sugar conversion (61.7%) from H. annuus biomass hydrolysate occurred, resulting in the production of 11.4 g/L of bioethanol. This is the first study to assess the suitability of phytoremediated sunflower biomass for bioethanol production.

Influence of different SSF conditions on ethanol production from corn stover at high solids loadings

DEFF Research Database (Denmark)

Gladis, Arne; Bondesson, Pia-Maria; Galbe, Mats

2015-01-01

In this study, three different kinds of simultaneous saccharification and fermentation (SSF) of washed pretreated corn stover with water-insoluble solids (WIS) content of 20% were investigated to find which one resulted in highest ethanol yield at high-solids loadings. The different methods were...

Effects of wet-pressing-induced fiber hornification on enzymatic saccharification of lignocelluloses

Science.gov (United States)

X.L. Luo; Junyong Zhu; Roland Gleisner; H.Y. Zhan

2011-01-01

This article reports the effect of wet-pressing-induced fiber hornification on enzymatic saccharification of lignocelluloses. A wet cellulosic substrate of bleached kraft eucalyptus pulp and two wet sulfite-pretreated lignocellulosic substrates of aspen and lodgepole pine were pressed to various moisture (solids) contents by variation of pressing pressure and pressing...

Electron beam irradiation pretreatment and enzymatic saccharification of used newsprint and paper mill wastes

International Nuclear Information System (INIS)

Khan, A.W.; Labrie, J.-P.; McKeown, Joseph

1987-01-01

Electron beam pretreatment of used newsprint, pulp, as well as pulp recovered from clarifier sludge and paper mill sludge, caused the dissociation of cellulose from lignin, and rendered them suitable for enzymatic hydrolysis. A maximum dose of 1 MGy for newsprint and 1.5-2.0 MGy for pulp and paper mill sludge was required to render cellulose present in them in a form which, could be enzymatically saccharified to 90% of completion. Saccharification approaching the theoretical yield was obtained in 2 days with a cellulolytic enzyme system obtained from Trichoderma reesei. As a result of irradiation, water soluble lignin breakdown products, NaOH- soluble lignin, free cellobiose, glucose, mannose, xylose and their polymers, and acetic acid were produced from these materials. (author)

Stability of cell wall composition and saccharification efficiency in Miscanthus across diverse environments

NARCIS (Netherlands)

Weijde, van der Tim; Dolstra, Oene; Visser, Richard G.F.; Trindade, Luisa M.

2017-01-01

To investigate the potential effects of differences between growth locations on the cell wall composition and saccharification efficiency of the bioenergy crop miscanthus, a diverse set of 15 accessions were evaluated in six locations across Europe for the first 3 years following establishment.

Xylanase supplementation on enzymatic saccharification of dilute acid pretreated poplars at different severities

Science.gov (United States)

Chao Zhang; Xinshu Zhuang; Zhao Jiang Wang; Fred Matt; Franz St. John; J.Y. Zhu

2013-01-01

Three pairs of solid substrates from dilute acid pretreatment of two poplar wood samples were enzymatically hydrolyzed by cellulase preparations supplemented with xylanase. Supplementation of xylanase improved cellulose saccharification perhaps due to improved cellulose accessibility by xylan hydrolysis. Total xylan removal directly affected enzymatic cellulose...

Dynamic Model-Based Evaluation of Process Configurations for Integrated Operation of Hydrolysis and Co-Fermentation for Bioethanol Production from Lignocellulose

DEFF Research Database (Denmark)

Morales Rodriguez, Ricardo; Meyer, Anne S.; Gernaey, Krist

2011-01-01

In this study a number of different process flowsheets were generated and their feasibility evaluated using simulations of dynamic models. A dynamic modeling framework was used for the assessment of operational scenarios such as, fed-batch, continuous and continuous with recycle configurations. E......) operating in continuous mode with a recycle of the SSCF reactor effluent, results in the best productivity of bioethanol among the proposed process configurations, with a yield of 0.18 kg ethanol/kg dry-biomass........ Each configuration was evaluated against the following benchmark criteria, yield (kg ethanol/kg dry-biomass), final product concentration and number of unit operations required in the different process configurations. The results show that simultaneous saccharification and co-fermentation (SSCF...

A thermophilic ionic liquid-tolerant cellulase cocktail for the production of cellulosic biofuels.

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Joshua I Park

Full Text Available Generation of biofuels from sugars in lignocellulosic biomass is a promising alternative to liquid fossil fuels, but efficient and inexpensive bioprocessing configurations must be developed to make this technology commercially viable. One of the major barriers to commercialization is the recalcitrance of plant cell wall polysaccharides to enzymatic hydrolysis. Biomass pretreatment with ionic liquids (ILs enables efficient saccharification of biomass, but residual ILs inhibit both saccharification and microbial fuel production, requiring extensive washing after IL pretreatment. Pretreatment itself can also produce biomass-derived inhibitory compounds that reduce microbial fuel production. Therefore, there are multiple points in the process from biomass to biofuel production that must be interrogated and optimized to maximize fuel production. Here, we report the development of an IL-tolerant cellulase cocktail by combining thermophilic bacterial glycoside hydrolases produced by a mixed consortia with recombinant glycoside hydrolases. This enzymatic cocktail saccharifies IL-pretreated biomass at higher temperatures and in the presence of much higher IL concentrations than commercial fungal cocktails. Sugars obtained from saccharification of IL-pretreated switchgrass using this cocktail can be converted into biodiesel (fatty acid ethyl-esters or FAEEs by a metabolically engineered strain of E. coli. During these studies, we found that this biodiesel-producing E. coli strain was sensitive to ILs and inhibitors released by saccharification. This cocktail will enable the development of novel biomass to biofuel bioprocessing configurations that may overcome some of the barriers to production of inexpensive cellulosic biofuels.

A Thermophilic Ionic Liquid-Tolerant Cellulase Cocktail for the Production of Cellulosic Biofuels

Science.gov (United States)

Park, Joshua I.; Steen, Eric J.; Burd, Helcio; Evans, Sophia S.; Redding-Johnson, Alyssa M.; Batth, Tanveer; Benke, Peter I.; D'haeseleer, Patrik; Sun, Ning; Sale, Kenneth L.; Keasling, Jay D.; Lee, Taek Soon; Petzold, Christopher J.; Mukhopadhyay, Aindrila; Singer, Steven W.; Simmons, Blake A.; Gladden, John M.

2012-01-01

Generation of biofuels from sugars in lignocellulosic biomass is a promising alternative to liquid fossil fuels, but efficient and inexpensive bioprocessing configurations must be developed to make this technology commercially viable. One of the major barriers to commercialization is the recalcitrance of plant cell wall polysaccharides to enzymatic hydrolysis. Biomass pretreatment with ionic liquids (ILs) enables efficient saccharification of biomass, but residual ILs inhibit both saccharification and microbial fuel production, requiring extensive washing after IL pretreatment. Pretreatment itself can also produce biomass-derived inhibitory compounds that reduce microbial fuel production. Therefore, there are multiple points in the process from biomass to biofuel production that must be interrogated and optimized to maximize fuel production. Here, we report the development of an IL-tolerant cellulase cocktail by combining thermophilic bacterial glycoside hydrolases produced by a mixed consortia with recombinant glycoside hydrolases. This enzymatic cocktail saccharifies IL-pretreated biomass at higher temperatures and in the presence of much higher IL concentrations than commercial fungal cocktails. Sugars obtained from saccharification of IL-pretreated switchgrass using this cocktail can be converted into biodiesel (fatty acid ethyl-esters or FAEEs) by a metabolically engineered strain of E. coli. During these studies, we found that this biodiesel-producing E. coli strain was sensitive to ILs and inhibitors released by saccharification. This cocktail will enable the development of novel biomass to biofuel bioprocessing configurations that may overcome some of the barriers to production of inexpensive cellulosic biofuels. PMID:22649505

Production of fructose-containing syrup with enzymes

Energy Technology Data Exchange (ETDEWEB)

Helwiig-Nielsen, B

1981-01-01

A review on enzymic processes used for production of fructose- high syrup from starch including liquefaction by alpha-amylase, saccharification by amyloglucosidase, and isomerization with glucose isomerase.

Wet oxidation pre-treatment of woody yard waste: Parameter optimization and enzymatic digestibility for ethanol production

DEFF Research Database (Denmark)

Lissens, G.; Klinke, H.B.; Verstraete, W.

2004-01-01

, 58-67% and 80-83% of the cellulose and hemicellulose contained in the waste were converted into monomeric sugars. The cellulose conversion efficiency during a simultaneous saccharification and fermentation (SSF) assay at 10% DM was 79% for the highest enzyme loading (25 FPU g(-1) DM) while 69...

Chemical Pretreatment-Independent Saccharifications of Xylan and Cellulose of Rice Straw by Bacterial Weak Lignin-Binding Xylanolytic and Cellulolytic Enzymes.

Science.gov (United States)

Teeravivattanakit, Thitiporn; Baramee, Sirilak; Phitsuwan, Paripok; Sornyotha, Somphit; Waeonukul, Rattiya; Pason, Patthra; Tachaapaikoon, Chakrit; Poomputsa, Kanokwan; Kosugi, Akihiko; Sakka, Kazuo; Ratanakhanokchai, Khanok

2017-11-15

demonstrates efficient synergism with endoxylanases PcXyn10C and PcXyn11A to depolymerize xylan in untreated rice straw and enhanced the xylose production and improved cellulose hydrolysis. Therefore, it can be considered an enzymatic pretreatment. Furthermore, the studies here show that glucose yield released from steam- and xylanolytic enzyme-treated rice straw by the combination of CtCel9R and TbCglT was higher than the glucose yield obtained from ammonia-treated rice straw saccharification. This work presents a novel environment-friendly xylanolytic enzyme pretreatment not only as a green pretreatment but also as an economically feasible biorefinery method. Copyright © 2017 American Society for Microbiology.

Evaluation of Alkali-Pretreated Soybean Straw for Lignocellulosic Bioethanol Production

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Seonghun Kim

2018-01-01

Full Text Available Soybean straw is a renewable resource in agricultural residues that can be used for lignocellulosic bioethanol production. To enhance enzymatic digestibility and fermentability, the biomass was prepared with an alkali-thermal pretreatment (sodium hydroxide, 121°C, 60 min. The delignification yield was 34.1~53%, in proportion to the amount of sodium hydroxide, from 0.5 to 3.0 M. The lignin and hemicellulose contents of the pretreated biomass were reduced by the pretreatment process, whereas the proportion of cellulose was increased. Under optimal condition, the pretreated biomass consisted of 74.0±0.1% cellulose, 10.3±0.1% hemicellulose, and 10.1±0.6% lignin. During enzymatic saccharification using Cellic® CTec2 cellulase, 10% (w/v of pretreated soybean straw was hydrolyzed completely and converted to 67.3±2.1 g/L glucose and 9.4±0.5 g/L xylose with a 90.9% yield efficiency. Simultaneous saccharification and fermentation of the pretreated biomass by Saccharomyces cerevisiae W303-1A produced 30.5±1.2 g/L ethanol in 0.5 L fermented medium containing 10% (w/v pretreated biomass after 72 h. The ethanol productivity was 0.305 g ethanol/g dry biomass and 0.45 g ethanol/g glucose after fermentation, with a low concentration of organic acid metabolites. Also, 82% of fermentable sugar was used by the yeast for ethanol fermentation. These results show that the combination of alkaline pretreatment and biomass hydrolysate is useful for enhancing bioethanol productivity using delignified soybean straw.

Enhancing saccharification of cassava stems by starch hydrolysis prior to pretreatment

OpenAIRE

Martín, Carlos; Wei, Maogui; Xiong, Shaojun; Jönsson, Leif J.

2017-01-01

Chemical characterization of cassava stems from different origin revealed that glucans accounted for 54-63% of the dry weight, whereas 35-67% of these glucans consisted of starch. The cassava stems were subjected to a saccharification study including starch hydrolysis, pretreatment with either sulfuric acid or 1-ethyl-3-methylimidazolium acetate ([Emim]OAc), and enzymatic hydrolysis of cellulose. Starch hydrolysis prior to pretreatment decreased sugar degradation, improved enzymatic convertib...

Studi Konsentrasi Amiloglukosidase dan Saccharomyces cereviseae dalam Produksi Bioetanol dari Ubi Jalar Melalui Sakarifikasi Fermentasi Simultan

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Bambang Admadi Harsojuwono

2015-03-01

Full Text Available The purpose of this study is to obtain the enzyme concentration and Sacharomyces cereviceae amyloglukosidase best to process simultaneous saccharification fermentation (SFS in the production of bio ethanol from sweet potato hydrolyzated. This study was designed using a factorial randomized block design. The first factor is the concentration of enzyme amiloglukosidase consisting of 3 levels ie 0.8; 1.0 and 1.2 ml / kg substrate. The second factor is the concentration of S. cereviceae consisting of 3 levels ie 5; 10 and 15% (v / v. Variable measured include of ethanol concentration, yield, efficiency of substrates using, fermentation efficiency and concentration of substrate consumption. Data were analyzed diversity and Duncan's multiple comparison test was done to determine the best treatment. The results showed that the concentration of the enzyme amyloglukosidase 1.2 ml / kg of substrate (3000 U / ml at a concentration of S. cereviceae 10% (v / v is the best treatment with the resulting ethanol concentration 7.48% (v / v, yield 19.89%, the efficiency of product formation by substrate 47.37%, 92.88% fermentation efficiency, and the concentration of substrate consumption of 15.78 g / L.

Bioprospecting thermophiles for cellulase production: a review.

Science.gov (United States)

Acharya, Somen; Chaudhary, Anita

2012-07-01

Most of the potential bioprospecting is currently related to the study of the extremophiles and their potential use in industrial processes. Recently microbial cellulases find applications in various industries and constitute a major group of industrial enzymes. Considerable amount of work has been done on microbial cellulases, especially with resurgence of interest in biomass ethanol production employing cellulases and use of cellulases in textile and paper industry. Most efficient method of lignocellulosic biomass hydrolysis is through enzymatic saccharification using cellulases. Significant information has also been gained about the physiology of thermophilic cellulases producers and process development for enzyme production and biomass saccharification. The review discusses the current knowledge on cellulase producing thermophilic microorganisms, their physiological adaptations and control of cellulase gene expression. It discusses the industrial applications of thermophilic cellulases, their cost of production and challenges in cellulase research especially in the area of improving process economics of enzyme production.

Bacterial laminarinase for application in ethanol production from brown algae Sargassum sp. using halotolerant yeast

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C.M.T. Perez

2018-03-01

Full Text Available Macroalgae are known to have many industrial applications, with current research targeting the potential of macroalgal biomass as feedstock in production of biofuels. Marine algal biomass is rich in storage carbohydrates, laminarin, and cellulose, which can be converted to fermentable sugars using appropriate enzymes, for fermentation to ethanol. This study focused on ethanol production from macroalgae using only enzymatic treatment for saccharification of algal biomass. This involved the isolation and identification of cellulase and laminarinase-producing microorganisms from mangrove area in the Philippines and production of partially purified enzymes for algal biomass saccharification. Results showed that the partially purified laminarinase produced from Bacillus sp. was capable of hydrolyzing the laminarin present in the macroalage. Fermentation of the algal hydrolysate yielded only small amount of ethanol due to lack of other pre-treatment methods, however, it was observed that higher ethanol was produced in saccharification treatments using a combination of cellulase and laminarinase which implies a possible synergistic effect between the two enzymes.

The Potential of Cellulosic Ethanol Production from Grasses in Thailand

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Jinaporn Wongwatanapaiboon

2012-01-01

Full Text Available The grasses in Thailand were analyzed for the potentiality as the alternative energy crops for cellulosic ethanol production by biological process. The average percentage composition of cellulose, hemicellulose, and lignin in the samples of 18 types of grasses from various provinces was determined as 31.85–38.51, 31.13–42.61, and 3.10–5.64, respectively. The samples were initially pretreated with alkaline peroxide followed by enzymatic hydrolysis to investigate the enzymatic saccharification. The total reducing sugars in most grasses ranging from 500–600 mg/g grasses (70–80% yield were obtained. Subsequently, 11 types of grasses were selected as feedstocks for the ethanol production by simultaneous saccharification and cofermentation (SSCF. The enzymes, cellulase and xylanase, were utilized for hydrolysis and the yeasts, Saccharomyces cerevisiae and Pichia stipitis, were applied for cofermentation at 35°C for 7 days. From the results, the highest yield of ethanol, 1.14 g/L or 0.14 g/g substrate equivalent to 32.72% of the theoretical values was obtained from Sri Lanka ecotype vetiver grass. When the yields of dry matter were included in the calculations, Sri Lanka ecotype vetiver grass gave the yield of ethanol at 1,091.84 L/ha/year, whereas the leaves of dwarf napier grass showed the maximum yield of 2,720.55 L/ha/year (0.98 g/L or 0.12 g/g substrate equivalent to 30.60% of the theoretical values.

Improving enzymatic saccharification of cassava stem using peroxide and microwave assisted pre-treatment techniques

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Sudha A.

2017-01-01

Full Text Available The effectiveness of microwave assisted alkali (MAA and alkaline hydrogen peroxide (AHP pre-treatment methods in improving the enzymatic saccharification of cassava stem was investigated. Ground cassava stems were by MAA method by varying microwave power, NaOH concentration and pre-treatment time. AHP method was performed at various H2O2 concentrations, pre-treatment temperatures and times. The results showed that reducing sugar yield was higher from MAA pretreated stem when compared with AHP pre-treatment, which demonstrated that MAA pre-treatment was effective in releasing sugars. SEM studies on the pre-treated samples revealed extensive distortion of fibres in MAA pre-treated than AHP pre-treated samples, which showed pores and cracks in the fibrous structure. Spectral studies showed the change in the chemical structure of pre-treated samples. The work revealed that the studied pre-treatment methods were effective in improving the enzymatic saccharification of cassava stem.

Biomass enzymatic saccharification is determined by the non-KOH-extractable wall polymer features that predominately affect cellulose crystallinity in corn.

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Jun Jia

Full Text Available Corn is a major food crop with enormous biomass residues for biofuel production. Due to cell wall recalcitrance, it becomes essential to identify the key factors of lignocellulose on biomass saccharification. In this study, we examined total 40 corn accessions that displayed a diverse cell wall composition. Correlation analysis showed that cellulose and lignin levels negatively affected biomass digestibility after NaOH pretreatments at p<0.05 & 0.01, but hemicelluloses did not show any significant impact on hexoses yields. Comparative analysis of five standard pairs of corn samples indicated that cellulose and lignin should not be the major factors on biomass saccharification after pretreatments with NaOH and H2SO4 at three concentrations. Notably, despite that the non-KOH-extractable residues covered 12%-23% hemicelluloses and lignin of total biomass, their wall polymer features exhibited the predominant effects on biomass enzymatic hydrolysis including Ara substitution degree of xylan (reverse Xyl/Ara and S/G ratio of lignin. Furthermore, the non-KOH-extractable polymer features could significantly affect lignocellulose crystallinity at p<0.05, leading to a high biomass digestibility. Hence, this study could suggest an optimal approach for genetic modification of plant cell walls in bioenergy corn.

Simultaneous action of external irradiation and products of air radiolysis

International Nuclear Information System (INIS)

Golikov, V.Ya.; Karklinskaya, O.N.; Mikhalev, V.P.; Vorotyntsev, A.P.; Kotov, N.N.

1977-01-01

To study the radiation damage due to the simultaneous effect of the acute ionizing radiation on the organism and the products of air radiolysis-ozone and nitrogen oxides, laboratory experiments were made. The scheme of the irradiation was the following: the acute dynamic effect (per hour) of X-rays at different doses, and nitrogen oxides and ozone with different concentrations. Male mice were exposed to irradiation (dose of 50-1000 rad, for 60 min.). Data obtained (mortality, survival, exponents of the peripheral blood) proved the radioprotective ability of the chemical agents under study. The radioprotective action was most pronounced at high concentrations. The fact proves that the mechanism of the simultaneous action of the products of air radiolysis and X-ray radiation is complex, and further efforts should be made at its investigation

Effect of Maize Biomass Composition on the Optimization of Dilute-Acid Pretreatments and Enzymatic Saccharification

NARCIS (Netherlands)

Torres Salvador, A.F.; Weijde, van der R.T.; Dolstra, O.; Visser, R.G.F.; Trindade, L.M.

2013-01-01

At the core of cellulosic ethanol research are innovations leading to reductions in the chemical and energetic stringency of thermochemical pretreatments and enzymatic saccharification. In this study, key compositional features of maize cell walls influencing the enzymatic conversion of biomass into

Comparison of microwave and conduction-convection heating autohydrolysis pretreatment for bioethanol production.

Science.gov (United States)

Aguilar-Reynosa, Alejandra; Romaní, Aloia; Rodríguez-Jasso, Rosa M; Aguilar, Cristóbal N; Garrote, Gil; Ruiz, Héctor A

2017-11-01

This work describes the application of two forms of heating for autohydrolysis pretreatment on isothermal regimen: conduction-convection heating and microwave heating processing using corn stover as raw material for bioethanol production. Pretreatments were performed using different operational conditions: residence time (10-50 min) and temperature (160-200°C) for both pretreatments. Subsequently, the susceptibility of pretreated solids was studied using low enzyme loads, and high substrate loads. The highest conversion was 95.1% for microwave pretreated solids. Also solids pretreated by microwave heating processing showed better ethanol conversion in simultaneous saccharification and fermentation process (92% corresponding to 33.8g/L). Therefore, microwave heating processing is a promising technology in the pretreatment of lignocellulosic materials. Copyright © 2017 Elsevier Ltd. All rights reserved.

Enhanced ethanol and glucosamine production from rice husk by NAOH pretreatment and fermentation by fungus Mucor hiemalis

Directory of Open Access Journals (Sweden)

Maryam Omidvar

2016-09-01

Full Text Available Ethanol production from rice husk by simultaneous saccharification and fermentation using Mucor hiemalis was investigated. To reach the maximum ethanol production yield, the most important influencing factors in the pretreatment process, including temperature (0-100°C, NaOH concentration (1-3 M, and the pretreatment time (30-180 min, were optimized using an experimental design by a response surface methodology (RSM. The maximum ethanol production yield of 86.7 % was obtained after fungal cultivation on the husk pretreated with 2.6 M NaOH at 67°C for 150 min. This was higher than the yield of 57.7% obtained using Saccharomyces cerevisiae as control. Furthermore, fermentation using M. hiemalis under the optimum conditions led to the production of a highly valuable fungal biomass, containing 60 g glucosamine (GlcN, 410 g protein, and 160 g fungal oil per each kg of the fungal biomass.

Lignosulfonate To Enhance Enzymatic Saccharification of Lignocelluloses: Role of Molecular Weight and Substrate Lignin

Science.gov (United States)

Haifeng Zhou; Hongming Lou; Dongjie Yang; J.Y. Zhu; Xueqing Qiu

2013-01-01

This study conducted an investigation of the effect of lignosulfonate (LS) on enzymatic saccharification of lignocelluloses. Two commercial LSs and one laboratory sulfonated kraft lignin were applied to Whatman paper, dilute acid and SPORL (sulfite pretreatment to overcome recalcitrance of lignocelluloses) pretreated aspen, and kraft alkaline and SPORL pretreated...

Fungal cellulases as an aid for the saccharification of cassava

Energy Technology Data Exchange (ETDEWEB)

De Menezes, T J.B.; Arakaki, T; DeLamo, P R; Sales, A M

1978-04-01

Culture broths of cellulolytic fungi were used together with commercial anylases to enhance the saccharification of cassava starch slurry. It was found that the addition of appropriate concentration of the cellulases Trichoderma viride and a soil isolated Basidiomycete, increased both the rate of sugar formation and the degree of solubilization, and decreased the viscosity of the hydrolyzates. Owing to the improvement of the rheological properties of the must, and the additional sugar produced, an increased ethanol yield would be expected from the alcoholic fermentation of this hydrolyzate.

Bioprospecting thermophiles for cellulase production: a review

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Somen Acharya

2012-09-01

Full Text Available Most of the potential bioprospecting is currently related to the study of the extremophiles and their potential use in industrial processes. Recently microbial cellulases find applications in various industries and constitute a major group of industrial enzymes. Considerable amount of work has been done on microbial cellulases, especially with resurgence of interest in biomass ethanol production employing cellulases and use of cellulases in textile and paper industry. Most efficient method of lignocellulosic biomass hydrolysis is through enzymatic saccharification using cellulases. Significant information has also been gained about the physiology of thermophilic cellulases producers and process development for enzyme production and biomass saccharification. The review discusses the current knowledge on cellulase producing thermophilic microorganisms, their physiological adaptations and control of cellulase gene expression. It discusses the industrial applications of thermophilic cellulases, their cost of production and challenges in cellulase research especially in the area of improving process economics of enzyme production.

Sequential high gravity ethanol fermentation and anaerobic digestion of steam explosion and organosolv pretreated corn stover.

Science.gov (United States)

Katsimpouras, Constantinos; Zacharopoulou, Maria; Matsakas, Leonidas; Rova, Ulrika; Christakopoulos, Paul; Topakas, Evangelos

2017-11-01

The present work investigates the suitability of pretreated corn stover (CS) to serve as feedstock for high gravity (HG) ethanol production at solids-content of 24wt%. Steam explosion, with and without the addition of H 2 SO 4 , and organosolv pretreated CS samples underwent a liquefaction/saccharification step followed by simultaneous saccharification and fermentation (SSF). Maximum ethanol concentration of ca. 76g/L (78.3% ethanol yield) was obtained from steam exploded CS (SECS) with 0.2% H 2 SO 4 . Organosolv pretreated CS (OCS) also resulted in high ethanol concentration of ca. 65g/L (62.3% ethanol yield). Moreover, methane production through anaerobic digestion (AD) was conducted from fermentation residues and resulted in maximum methane yields of ca. 120 and 69mL/g volatile solids (VS) for SECS and OCS samples, respectively. The results indicated that the implementation of a liquefaction/saccharification step before SSF employing a liquefaction reactor seemed to handle HG conditions adequately. Copyright © 2017 Elsevier Ltd. All rights reserved.

Hydrodynamic cavitation as a novel pretreatment approach for bioethanol production from reed.

Science.gov (United States)

Kim, Ilgook; Lee, Ilgyu; Jeon, Seok Hwan; Hwang, Taewoon; Han, Jong-In

2015-09-01

In this study, hydrodynamic cavitation (HC) was employed as a physical means to improve alkaline pretreatment of reed. The HC-assisted alkaline pretreatment was undertaken to evaluate the influence of NaOH concentration (1-5%), solid-to-liquid ratio (5-15%), and reaction time (20-60 min) on glucose yield. The optimal condition was found to be 3.0% NaOH at solid-to-liquid (S/L) ratio of 11.8% for 41.1 min, which resulted in the maximum glucose yield of 326.5 g/kg biomass. Furthermore, simultaneous saccharification and fermentation (SSF) was conducted to assess the ethanol production. An ethanol concentration of 25.9 g/L and ethanol yield of 90% were achieved using batch SSF. These results clearly demonstrated HC system can be indeed a promising pretreatment tool for lignocellulosic bioethanol production. Copyright © 2015 Elsevier Ltd. All rights reserved.

Browse Title Index

African Journals Online (AJOL)

Items 851 - 900 of 1305 ... Vol 9, No 1 (2005), Optimal design of wastewater treatment plant using adaptive simulated annealing, Abstract. L Govindarajan, Kumar Krishna, T Karunanithi. Vol 22, No 1 (2018), Optimization of bioethanol production from simultaneous saccharification and fermentation of pineapple peels using ...

Distortion product otoacoustic emissions: comparison of sequential vs. simultaneous presentation of primary tones.

Science.gov (United States)

Kumar, U Ajith; Maruthy, Sandeep; Chandrakant, Vishwakarma

2009-03-01

Distortion product otoacoustic emissions are one form of evoked otoacoustic emissions. DPOAEs provide the frequency specific information about the hearing status in mid and high frequency regions. But in most screening protocols TEOAEs are preferred as it requires less time compared to DPOAE. This is because, in DPOAE each stimulus is presented one after the other and responses are analyzed. Grason and Stadler Incorporation 60 (GSI-60) offer simultaneous presentation of four sets of primary tones at a time and checks for the DPOAE. In this mode of presentation, all the pairs are presented at a time and following that response is extracted separately whereas, in sequential mode primaries are presented in orderly fashion one after the other. In this article simultaneous and sequential protocols were used to compare the Distortion product otoacoustic emission amplitude, noise floor and administration time in individuals with normal hearing and mild sensori-neural (SN) hearing loss. In simultaneous protocols four sets of primary tones (i.e. 8 tones) were presented together whereas, in sequential presentation mode one set of primary tones was presented each time. Simultaneous protocol was completed in less than half the time required for the completion of sequential protocol. Two techniques yielded similar results at frequencies above 1000 Hz only in normal hearing group. In SN hearing loss group simultaneous presentation yielded signifi cantly higher noise floors and distortion product amplitudes. This result challenges the use of simultaneous presentation technique in neonatal hearing screening programmes and on other pathologies. This discrepancy between two protocols may be due to some changes in biomechanical process in the cochlear and/or due to higher distortion/noise produced by the system during the simultaneous presentation mode.

Production of bioethanol from corn meal hydrolyzates

Energy Technology Data Exchange (ETDEWEB)

Ljiljana Mojovic; Svetlana Nikolic; Marica Rakin; Maja Vukasinovic [University of Belgrade, Belgrade (Serbia and Montenegro). Faculty of Technology and Metallurgy, Department of Biochemical Engineering and Biotechnology

2006-09-15

The two-step enzymatic hydrolysis of corn meal by commercially available {alpha}-amylase and glucoamylase and further ethanol fermentation of the obtained hydrolyzates by Saccharomyces cerevisiae yeast was studied. The conditions of starch hydrolysis such as substrate and enzyme concentration and the time required for enzymatic action were optimized taking into account both the effects of hydrolysis and ethanol fermentation. The corn meal hydrolyzates obtained were good substrates for ethanol fermentation by S. cerevisiae. The yield of ethanol of more than 80% (w/w) of the theoretical was achieved with a satisfactory volumetric productivity P (g/l h). No shortage of fermentable sugars was observed during simultaneous hydrolysis and fermentation. In this process, the savings in energy by carrying out the saccharification step at lower temperature (32{sup o}C) could be realized, as well as a reduction of the process time for 4 h. 31 refs., 5 figs., 2 tabs.

Comparison of several ethanol productions using xylanase, inorganic salts, surfactant

Science.gov (United States)

Wu, Yan; Lu, Jie; Yang, Rui-feng; Song, Wen-jing; Li, Hai-ming; Wang, Hai-song; Zhou, Jing-hui

2017-03-01

Liquid hot water (LHW) pretreatment is an effective and environmentally friendly method to produce bioethanol with lignocellulosic materials. Corn stover was pretreated with liquid hot water (LHW) and then subjected to semi-simultaneous saccharification and fermentation (S-SSF) to obtain high ethanol concentration and yield. The present study aimed to confirm the effect of several additives on the fermentation digestibility of unwashed WIS of corn stover pretreated with LHW. So we also investigated the process, such as enzyme addition, inorganic salts, surfactant and different loading Triton. Results show that high ethanol concentration is necessary to add xylanase in the stage of saccharification. The ethanol concentration increased mainly with magnesium ion on fermentation. Comparing with Tween 80, Span 80 and Polyethylene glycol, Triton is the best surfactant. In contrast to using xylanase and Triton respectively, optimization can make up the lack of stamina and improve effect of single inorganic salts.

Comparison of Dilute Acid and Ionic Liquid Pretreatment of Switchgrass: Biomass Recalcitrance, Delignification and Enzymatic Saccharification

Science.gov (United States)

The efficiency of two biomass pretreatment technologies, dilute acid hydrolysis and dissolution in an ionic liquid, are compared in terms of delignification, saccharification efficiency and saccharide yields with switchgrass serving as a model bioenergy crop. When subject to ionic liquid pretreatme...

Impact of Different Lignin Fractions on Saccharification Efficiency in Diverse Species of the Bioenergy Crop Miscanthus

NARCIS (Netherlands)

Weijde, van der Tim; Torres Salvador, Andres Francisco; Dolstra, Oene; Dechesne, Annemarie; Visser, Richard G.F.; Trindade, Luisa M.

2016-01-01

Lignin is a key factor limiting saccharification of lignocellulosic feedstocks. In this comparative study, various lignin methods—including acetyl bromide lignin (ABL), acid detergent lignin (ADL), Klason lignin (KL), and modified ADL and KL determination methods—were evaluated for their

Wet oxidation pretreatment of rape straw for ethanol production

DEFF Research Database (Denmark)

Arvaniti, Efthalia; Bjerre, Anne Belinda; Schmidt, Jens Ejbye

2012-01-01

Rape straw can be used for production of second generation bioethanol. In this paper we optimized the pretreatment of rape straw for this purpose using Wet oxidation (WO). The effect of reaction temperature, reaction time, and oxygen gas pressure was investigated for maximum ethanol yield via...... Simultaneous Saccharification and Fermentation (SSF). To reduce the water use and increase the energy efficiency in WO pretreatment features like recycling liquid (filtrate), presoaking of rape straw in water or recycled filtrate before WO, skip washing pretreated solids (filter cake) after WO, or use of whole...... gas produced higher ethanol yields and cellulose, hemicelluloses, and lignin recoveries, than 15 min WO treatment at 195 °C. Also, recycling filtrate and use of higher oxygen gas pressure reduced recovery of materials. The use of filtrate could be inhibitory for the yeast, but also reduced lactic acid...

Steam-exploded biomass saccharification is predominately affected by lignocellulose porosity and largely enhanced by Tween-80 in Miscanthus.

Science.gov (United States)

Sun, Dan; Alam, Aftab; Tu, Yuanyuan; Zhou, Shiguang; Wang, Yanting; Xia, Tao; Huang, Jiangfeng; Li, Ying; Zahoor; Wei, Xiaoyang; Hao, Bo; Peng, Liangcai

2017-09-01

In this study, total ten Miscanthus accessions exhibited diverse cell wall compositions, leading to largely varied hexoses yields at 17%-40% (% cellulose) released from direct enzymatic hydrolysis of steam-exploded (SE) residues. Further supplied with 2% Tween-80 into the enzymatic digestion, the Mis7 accession showed the higher hexose yield by 14.8-fold than that of raw material, whereas the Mis10 had the highest hexoses yield at 77% among ten Miscanthus accessions. Significantly, this study identified four wall polymer features that negatively affect biomass saccharification as pbiomass enzymatic digestion. Hence, this study provides the potential strategy to enhance biomass saccharification using optimal biomass process technology and related genetic breeding in Miscanthus and beyond. Copyright © 2017 Elsevier Ltd. All rights reserved.

Ethanol production from wet oxidized corn straw by simultaneous saccharification and fermentation

DEFF Research Database (Denmark)

Zhang, Q.; Yin, Y.; Thygesen, Anders

2010-01-01

remained in the solid fraction and recovery of cellulose was 95.87% after pretreatment. After 24 h hydrolysis at 50°C using cellulase, the achieved conversion of cellulose to glucose was about 67.6%. After 142 h of SSF with substrate concentration of 8%, ethanol yield of 79.0% of the theoretical...

Enhancing saccharification of wheat straw by mixing enzymes from genetically-modified Trichoderma reesei and Aspergillus niger.

Science.gov (United States)

Jiang, Yanping; Duarte, Alexandra Vivas; van den Brink, Joost; Wiebenga, Ad; Zou, Gen; Wang, Chengshu; de Vries, Ronald P; Zhou, Zhihua; Benoit, Isabelle

2016-01-01

To increase the efficiency of enzymatic hydrolysis for plant biomass conversion into renewable biofuel and chemicals. By overexpressing the point mutation A824 V transcriptional activator Xyr1 in Trichoderma reesei, carboxymethyl cellulase, cellobiosidase and β-D-glucosidase activities of the best mutant were increased from 1.8 IU/ml, 0.1 IU/ml and 0.05 IU/ml to 4.8 IU/ml, 0.4 IU/ml and 0.3 IU/ml, respectively. The sugar yield of wheat straw saccharification by combining enzymes from this mutant and the Aspergillus niger genetically modified strain ΔcreA/xlnR c/araR c was improved up to 7.5 mg/ml, a 229 % increase compared to the combination of wild type strains. Mixing enzymes from T. reesei and A. niger combined with the genetic modification of transcription factors is a promising strategy to increase saccharification efficiency.

Ethanol production from Sorghum bicolor using both separate and ...

African Journals Online (AJOL)

STORAGESEVER

2009-06-17

Jun 17, 2009 ... pre-treatment, enzymatic saccharification, detoxification of inhibitors and fermentation of Sorghum bicolor straw for ethanol production ..... The authors wish to acknowledge financial support from ... Official energy statistics from.

Comparative study of sulfite pretreatments for robust enzymatic saccharification of corn cob residue

Directory of Open Access Journals (Sweden)

Bu Lingxi

2012-12-01

Full Text Available Abstract Background Corn cob residue (CCR is a kind of waste lignocellulosic material with enormous potential for bioethanol production. The moderated sulphite processes were used to enhance the hydrophily of the material by sulfonation and hydrolysis. The composition, FT-IR spectra, and conductometric titrations of the pretreated materials were measured to characterize variations of the CCR in different sulfite pretreated environments. And the objective of this study is to compare the saccharification rate and yield of the samples caused by these variations. Results It was found that the lignin in the CCR (43.2% had reduced to 37.8%, 38.0%, 35.9%, and 35.5% after the sulfite pretreatment in neutral, acidic, alkaline, and ethanol environments, respectively. The sulfite pretreatments enhanced the glucose yield of the CCR. Moreover, the ethanol sulfite sample had the highest glucose yield (81.2%, based on the cellulose in the treated sample among the saccharification samples, which was over 10% higher than that of the raw material (70.6%. More sulfonic groups and weak acid groups were produced during the sulfite pretreatments. Meanwhile, the ethanol sulfite treated sample had the highest sulfonic group (0.103 mmol/g and weak acid groups (1.85 mmol/g in all sulfite treated samples. In FT-IR spectra, the variation of bands at 1168 and 1190 cm-1 confirmed lignin sulfonation during sulfite pretreatment. The disappearance of the band at 1458 cm-1 implied the methoxyl on lignin had been removed during the sulfite pretreatments. Conclusions It can be concluded that the lignin in the CCR can be degraded and sulfonated during the sulfite pretreatments. The pretreatments improve the hydrophility of the samples because of the increase in sulfonic group and weak acid groups, which enhances the glucose yield of the material. The ethanol sulfite pretreatment is the best method for lignin removal and with the highest glucose yield.

production of bioethanol from rice straw using yeast extracts ...

African Journals Online (AJOL)

user

70% of production cost using less important materials, like agricultural waste ... rice cultivation and wood industries results in the ... method for pretreatment and enzymatic saccharification ... The economic problems consist exclusive of cost.

The relation between xyr1 overexpression in Trichoderma harzianum and sugarcane bagasse saccharification performance.

Science.gov (United States)

da Silva Delabona, Priscila; Rodrigues, Gisele Nunes; Zubieta, Mariane Paludetti; Ramoni, Jonas; Codima, Carla Aloia; Lima, Deise Juliana; Farinas, Cristiane Sanchez; da Cruz Pradella, José Geraldo; Seiboth, Bernhard

2017-03-20

This work investigates the influence of the positive regulator XYR1 of Trichoderma harzianum on the production of cellulolytic enzymes, using sugarcane bagasse as carbon source. Constitutive expression of xyr1 was achieved under the control of the strong Trichoderma reesei pki1 promoter. Five clones with xyr1 overexpression achieved higher xyr1 expression and greater enzymatic productivity when cultivated under submerged fermentation, hence validating the genetic construction for T. harzianum. Clone 5 presented a relative expression of xyr1 26-fold higher than the parent strain and exhibited 66, 37, and 36% higher values for filter paper activity, xylanase activity, and β-glucosidase activity, respectively, during cultivation in a stirred-tank bioreactor. The overexpression of xyr1 in T. harzianum resulted in an enzymatic complex with significantly improved performance in sugarcane bagasse saccharification, with an enhancement of 25% in the first 24h. Our results also show that constitutive overexpression of xyr1 leads to the induction of several important players in biomass degradation at early (24h) and also late (48h) timepoints of inoculation. However, we also observed that the carbon catabolite repressor CRE1 was upregulated in xyr1 overexpression mutants. These findings demonstrate the feasibility of improving cellulase production by modifying regulator expression and suggest an attractive approach for increasing total cellulase productivity in T. harzianum. Copyright © 2017 Elsevier B.V. All rights reserved.

Microwave-Assisted Alkali Pre-Treatment, Densification and Enzymatic Saccharification of Canola Straw and Oat Hull.

Science.gov (United States)

Agu, Obiora S; Tabil, Lope G; Dumonceaux, Tim

2017-03-26

The effects of microwave-assisted alkali pre-treatment on pellets' characteristics and enzymatic saccharification for bioethanol production using lignocellulosic biomass of canola straw and oat hull were investigated. The ground canola straw and oat hull were immersed in distilled water, sodium hydroxide and potassium hydroxide solutions at two concentrations (0.75% and 1.5% w/v) and exposed to microwave radiation at power level 713 W and three residence times (6, 12 and 18 min). Bulk and particle densities of ground biomass samples were determined. Alkaline-microwave pre-treated and untreated samples were subjected to single pelleting test in an Instron universal machine, pre-set to a load of 4000 N. The measured parameters, pellet density, tensile strength and dimensional stability were evaluated and the results showed that the microwave-assisted alkali pre-treated pellets had a significantly higher density and tensile strength compared to samples that were untreated or pre-treated by microwave alone. The chemical composition analysis showed that microwave-assisted alkali pre-treatment was able to disrupt and break down the lignocellulosic structure of the samples, creating an area of cellulose accessible to cellulase reactivity. The best enzymatic saccharification results gave a high glucose yield of 110.05 mg/g dry sample for canola straw ground in a 1.6 mm screen hammer mill and pre-treated with 1.5% NaOH for 18 min, and a 99.10 mg/g dry sample for oat hull ground in a 1.6 mm screen hammer mill and pre-treated with 0.75% NaOH for 18 min microwave-assisted alkali pre-treatments. The effects of pre-treatment results were supported by SEM analysis. Overall, it was found that microwave-assisted alkali pre-treatment of canola straw and oat hull at a short residence time enhanced glucose yield.

Inhibitory effects of phenolic compounds of rice straw formed by saccharification during ethanol fermentation by Pichia stipitis.

Science.gov (United States)

Wang, Xiahui; Tsang, Yiu Fai; Li, Yuhao; Ma, Xiubing; Cui, Shouqing; Zhang, Tian-Ao; Hu, Jiajun; Gao, Min-Tian

2017-11-01

In this study, it was found that the type of phenolic acids derived from rice straw was the major factor affecting ethanol fermentation by Pichia stipitis. The aim of this study was to investigate the inhibitory effect of phenolic acids on ethanol fermentation with rice straw. Different cellulases produced different ratios of free phenolic acids to soluble conjugated phenolic acids, resulting in different fermentation efficiencies. Free phenolic acids exhibited much higher inhibitory effect than conjugated phenolic acids. The flow cytometry results indicated that the damage to cell membranes was the primary mechanism of inhibition of ethanol fermentation by phenolic acids. The removal of free phenolic acids from the hydrolysates increased ethanol productivity by 2.0-fold, indicating that the free phenolic acids would be the major inhibitors formed during saccharification. The integrated process for ethanol and phenolic acids may constitute a new strategy for the production of low-cost ethanol. Copyright © 2017 Elsevier Ltd. All rights reserved.

Efficient production of ethanol from waste paper and the biochemical methane potential of stillage eluted from ethanol fermentation.

Science.gov (United States)

Nishimura, Hiroto; Tan, Li; Sun, Zhao-Yong; Tang, Yue-Qin; Kida, Kenji; Morimura, Shigeru

2016-02-01

Waste paper can serve as a feedstock for ethanol production due to being rich in cellulose and not requiring energy-intensive thermophysical pretreatment. In this study, an efficient process was developed to convert waste paper to ethanol. To accelerate enzymatic saccharification, pH of waste paper slurry was adjusted to 4.5-5.0 with H2SO4. Presaccharification and simultaneous saccharification and fermentation (PSSF) with enzyme loading of 40 FPU/g waste paper achieved an ethanol yield of 91.8% and productivity of 0.53g/(Lh) with an ethanol concentration of 32g/L. Fed-batch PSSF was used to decrease enzyme loading to 13 FPU/g waste paper by feeding two separate batches of waste paper slurry. Feeding with 20% w/w waste paper slurry increased ethanol concentration to 41.8g/L while ethanol yield decreased to 83.8%. To improve the ethanol yield, presaccharification was done prior to feeding and resulted in a higher ethanol concentration of 45.3g/L, a yield of 90.8%, and productivity of 0.54g/(Lh). Ethanol fermentation recovered 33.2% of the energy in waste paper as ethanol. The biochemical methane potential of the stillage eluted from ethanol fermentation was 270.5mL/g VTS and 73.0% of the energy in the stillage was recovered as methane. Integrating ethanol fermentation with methane fermentation, recovered a total of 80.4% of the energy in waste paper as ethanol and methane. Copyright © 2015 Elsevier Ltd. All rights reserved.

OsCESA9 conserved-site mutation leads to largely enhanced plant lodging resistance and biomass enzymatic saccharification by reducing cellulose DP and crystallinity in rice.

Science.gov (United States)

Li, Fengcheng; Xie, Guosheng; Huang, Jiangfeng; Zhang, Ran; Li, Yu; Zhang, Miaomiao; Wang, Yanting; Li, Ao; Li, Xukai; Xia, Tao; Qu, Chengcheng; Hu, Fan; Ragauskas, Arthur J; Peng, Liangcai

2017-09-01

Genetic modification of plant cell walls has been posed to reduce lignocellulose recalcitrance for enhancing biomass saccharification. Since cellulose synthase (CESA) gene was first identified, several dozen CESA mutants have been reported, but almost all mutants exhibit the defective phenotypes in plant growth and development. In this study, the rice (Oryza sativa) Osfc16 mutant with substitutions (W481C, P482S) at P-CR conserved site in CESA9 shows a slightly affected plant growth and higher biomass yield by 25%-41% compared with wild type (Nipponbare, a japonica variety). Chemical and ultrastructural analyses indicate that Osfc16 has a significantly reduced cellulose crystallinity (CrI) and thinner secondary cell walls compared with wild type. CESA co-IP detection, together with implementations of a proteasome inhibitor (MG132) and two distinct cellulose inhibitors (Calcofluor, CGA), shows that CESA9 mutation could affect integrity of CESA4/7/9 complexes, which may lead to rapid CESA proteasome degradation for low-DP cellulose biosynthesis. These may reduce cellulose CrI, which improves plant lodging resistance, a major and integrated agronomic trait on plant growth and grain production, and enhances biomass enzymatic saccharification by up to 2.3-fold and ethanol productivity by 34%-42%. This study has for the first time reported a direct modification for the low-DP cellulose production that has broad applications in biomass industries. © 2017 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

[Optimization of fuel ethanol production from kitchen waste by Plackett-Burman design].

Science.gov (United States)

Ma, Hong-Zhi; Gong, Li-Juan; Wang, Qun-Hui; Zhang, Wen-Yu; Xu, Wen-Long

2008-05-01

Kitchen garbage was chosen to produce ethanol through simultaneous saccharification and fermentation (SSF) by Zymomonas mobilis. Plackett-Burman design was employed to screen affecting parameters during SSF process. The parameters were divided into two parts, enzymes and nutritions. None of the nutritions added showed significant effect during the experiment, which demonstrated that the kitchen garbage could meet the requirement of the microorganism without extra supplementation. Protease and glucoamylase were determined to be affecting factors for ethanol production. Single factor experiment showed that the optimum usage of these two enzymes were both 100 U/g and the corresponding maximum ethanol was determined to be 53 g/L. The ethanol yield could be as high as 44%. The utilization of kitchen garbage to produce ethanol could reduce threaten of waste as well as improve the protein content of the spent. This method could save the ethanol production cost and benefit for the recycle of kitchen garbage.

Production of Bioethanol from Carrot Pomace Using the Thermotolerant Yeast Kluyveromyces marxianus

Energy Technology Data Exchange (ETDEWEB)

Chi-Yang Yu; Bo-Hong Jiang; Kow-Jen Duan [Tatung University, Tapei, Taiwan (China). Department of Bioengineering

2013-03-15

Carrot pomace, a major agricultural waste from the juice industry, was used as a feedstock for bioethanol production by fermentation with the thermotolerant yeast Kluyveromyces marxianus. Treatment of the carrot pomace with Accellerase(TM) 1000 and pectinase at 50 °C for 84 h, resulted in conversion of 42% of its mass to fermentable sugars, mainly glucose, fructose, and sucrose. Simultaneous saccharification and fermentation (SSF) at 42 °C was performed on 10% (w/v) carrot pomace; the concentration of ethanol reached 18 g/L and the yield of ethanol from carrot pomace was 0.18 g/g. The highest ethanol concentration of 37 g/L was observed with an additional charge of 10% supplemented to the original 10% of carrot pomace after 12 h; the corresponding yield was 0.185 g/g. Our results clearly demonstrated the potential of combining a SSF process with thermotolerant yeast for the production of bioethanol using carrot pomace as a feedstock.

Direct ethanol production from cellulosic materials at high temperature using the thermotolerant yeast Kluyveromyces marxianus displaying cellulolytic enzymes

Energy Technology Data Exchange (ETDEWEB)

Yanase, Shuhei; Yamada, Ryosuke; Ogino, Chiaki; Kondo, Akihiko [Kobe Univ. (Japan). Dept. of Chemical Science and Engineering; Hasunuma, Tomohisa; Tanaka, Tsutomu; Fukuda, Hideki [Kobe Univ. (Japan). Organization of Advanced Science and Technology

2010-09-15

To exploit cellulosic materials for fuel ethanol production, a microorganism capable of high temperature and simultaneous saccharification-fermentation has been required. However, a major drawback is the optimum temperature for the saccharification and fermentation. Most ethanol-fermenting microbes have an optimum temperature for ethanol fermentation ranging between 28 C and 37 C, while the activity of cellulolytic enzymes is highest at around 50 C and significantly decreases with a decrease in temperature. Therefore, in the present study, a thermotolerant yeast, Kluyveromyces marxianus, which has high growth and fermentation at elevated temperatures, was used as a producer of ethanol from cellulose. The strain was genetically engineered to display Trichoderma reesei endoglucanase and Aspergillus aculeatus {beta}-glucosidase on the cell surface, which successfully converts a cellulosic {beta}-glucan to ethanol directly at 48 C with a yield of 4.24 g/l from 10 g/l within 12 h. The yield (in grams of ethanol produced per gram of {beta}-glucan consumed) was 0.47 g/g, which corresponds to 92.2% of the theoretical yield. This indicates that high-temperature cellulose fermentation to ethanol can be efficiently accomplished using a recombinant K. marxianus strain displaying thermostable cellulolytic enzymes on the cell surface. (orig.)

ETHANOL ORGANOSOLV PRETREATMENT OF BAMBOO FOR EFFICIENT ENZYMATIC SACCHARIFICATION

Directory of Open Access Journals (Sweden)

Zhiqiang Li,

2012-06-01

Full Text Available Bamboo is a potential lignocellulosic biomass for the production of bioethanol because of its high cellulose and hemicelluloses content. In this research, ethanol organosolv pretreatment with dilute sulfuric acid as the catalyst was studied in order to enhance enzymatic saccharification of moso bamboo. The addition of 2% (w/w bamboo dilute sulfuric acid in 75% ethanol had a particularly strong effect on fractionation of bamboo. It yielded a solids fraction containing 83.4% cellulose in the treated substrate. The cellulose conversion to glucose yield reached 77.1 to 83.4% after enzymatic hydrolysis of the solids fraction for 48 h at an enzyme loading of 15 FPU cellulase/g cellulose and 30 IU β-glucosidase/g cellulose. The enzymatic hydrolysis rate was significantly accelerated as the ethanol organosolv pretreatment time increased, reaching the highest enzymatic glucose yield of 83.4% after 48 h at 50 °C. The concentrations of fermentation inhibitors such as HMF (5-hydroxy-2-methyl furfural and furfural were 0.96 g/L and 4.38 g/L in the spent liquor after the ethanol organosolv pretreatment, which were slightly lower than the concentrations quantified during H2SO4-water treatment. Spent liquor was diluted with water, and more than 87.2% of lignin in raw bamboo was recovered as ethanol organosolv lignin through the filtration process.

Ethanol production from a biomass mixture of furfural residues with green liquor-peroxide saccarified cassava liquid.

Science.gov (United States)

Ji, Li; Zheng, Tianran; Zhao, Pengxiang; Zhang, Weiming; Jiang, Jianxin

2016-06-01

As the most abundant renewable resources, lignocellulosic materials are ideal candidates as alternative feedstock for bioethanol production. Cassava residues (CR) are byproducts of the cassava starch industry which can be mixed with lignocellulosic materials for ethanol production. The presence of lignin in lignocellulosic substrates can inhibit saccharification by reducing the cellulase activity. Simultaneous saccharification and fermentation (SSF) of furfural residues (FR) pretreated with green liquor and hydrogen peroxide (GL-H2O2) with CR saccharification liquid was investigated. The final ethanol concentration, yield, initial rate, number of live yeast cells, and the dead yeast ratio were compared to evaluate the effectiveness of combining delignificated lignocellulosic substrates and starchy substrates for ethanol production. Our results indicate that 42.0 % of FR lignin removal was achieved on FR using of 0.06 g H2O2/g-substrate and 9 mL GL/g-substrate at 80 °C. The highest overall ethanol yield was 93.6 % of the theoretical. When the ratio of 0.06 g/g-H2O2-GL-pretreated FR to CR was 5:1, the ethanol concentration was the same with that ratio of untreated FR to CR of 1:1. Using 0.06 g/g-H2O2-GL-pretreated FR with CR at a ratio of 2:1 resulted in 51.9 g/L ethanol concentration. Moreover, FR pretreated with GL-H2O2 decreased the concentration of byproducts in SSF compared with that obtained in the previous study. The lignin in FR would inhibit enzyme activity and GL-H2O2 is an advantageous pretreatment method to treat FR and high intensity of FR pretreatment increased the final ethanol concentration. The efficiency of ethanol fermentation of was improved when delignification increased. GL-H2O2 is an advantageous pretreatment method to treat FR. As the pretreatment dosage of GL-H2O2 on FR increased, the proportion of lignocellulosic substrates was enhanced in the SSF of the substrate mixture of CR and FR as compared with untreated FR. Moreover, the

Saccharification of Agricultural Lignocellulose Feedstocks and Protein-Level Responses by a Termite Gut-Microbe Bioreactor

International Nuclear Information System (INIS)

Rajarapu, Swapna Priya; Scharf, Michael E.

2017-01-01

This study investigated saccharification and protein-level responses to the candidate biofuel feedstocks corn stover (CS) and soybean residue (SR) by the gut of a lower termite. The focus termite was Reticulitermes flavipes, which is a highly efficient digester of wood lignocellulose that houses a mixture of prokaryotic and eukaryotic microbes in its gut. Our specific objectives were to (i) measure saccharification potential of the CS and SR feedstocks by termite gut protein extracts, (ii) identify specific proteins in the termite gut responding to feeding on CS and SR diets, and (iii) evaluate gut lignocellulase and accessory enzyme activity responses to CS and SR feeding. Cellulose paper was the control diet. Although CS was saccharified at higher levels, termite gut protein extracts saccharified both CS and SR irrespective of feedstock loading. Consumption of the CS and SR feedstocks by termites resulted in surprisingly few differences in gut protein profiles, with the main exception being elevated myosin abundance with SR feeding. Activity of potential lignocellulases and accessory enzymes was generally similar between CS and SR fed guts as well; however, cellobiohydrolase/exoglucanase activity was higher with CS feeding and glutathione peroxidase activity with SR feeding. These findings have significance from two perspectives. First, SR feeding/digestion appears to cause physiological stress in the termite gut that likely would extend to other types of microbial environments including those within industrial bioreactors. Second, because termites can survive on exclusive CS and SR diets and their guts exhibit clear CS and SR saccharification activity, this validates the R. flavipes system as a potential source for CS and SR degrading enzymes; in particular, cellobiohydrolases/exoglucanases and glutathione peroxidases from this system may play roles in CS and SR breakdown.

Saccharification of Agricultural Lignocellulose Feedstocks and Protein-Level Responses by a Termite Gut-Microbe Bioreactor

Energy Technology Data Exchange (ETDEWEB)

Rajarapu, Swapna Priya; Scharf, Michael E., E-mail: mscharf@purdue.edu [Department of Entomology, Purdue University, West Lafayette, IN (United States)

2017-04-07

This study investigated saccharification and protein-level responses to the candidate biofuel feedstocks corn stover (CS) and soybean residue (SR) by the gut of a lower termite. The focus termite was Reticulitermes flavipes, which is a highly efficient digester of wood lignocellulose that houses a mixture of prokaryotic and eukaryotic microbes in its gut. Our specific objectives were to (i) measure saccharification potential of the CS and SR feedstocks by termite gut protein extracts, (ii) identify specific proteins in the termite gut responding to feeding on CS and SR diets, and (iii) evaluate gut lignocellulase and accessory enzyme activity responses to CS and SR feeding. Cellulose paper was the control diet. Although CS was saccharified at higher levels, termite gut protein extracts saccharified both CS and SR irrespective of feedstock loading. Consumption of the CS and SR feedstocks by termites resulted in surprisingly few differences in gut protein profiles, with the main exception being elevated myosin abundance with SR feeding. Activity of potential lignocellulases and accessory enzymes was generally similar between CS and SR fed guts as well; however, cellobiohydrolase/exoglucanase activity was higher with CS feeding and glutathione peroxidase activity with SR feeding. These findings have significance from two perspectives. First, SR feeding/digestion appears to cause physiological stress in the termite gut that likely would extend to other types of microbial environments including those within industrial bioreactors. Second, because termites can survive on exclusive CS and SR diets and their guts exhibit clear CS and SR saccharification activity, this validates the R. flavipes system as a potential source for CS and SR degrading enzymes; in particular, cellobiohydrolases/exoglucanases and glutathione peroxidases from this system may play roles in CS and SR breakdown.

Cost-effective production of cellulose hydrolysing enzymes from Trichoderma sp. RCK65 under SSF and its evaluation in saccharification of cellulosic substrates.

Science.gov (United States)

Chakraborty, Subhojit; Gupta, Rishi; Jain, Kavish Kumar; Kuhad, Ramesh Chander

2016-11-01

Trichoderma sp. is a potential cellulase producing mesophilic fungi which grow under mild acidic condition. In this study, growth and nutritional conditions were manipulated for the maximum and cost-effective production of cellulase using lab strain Trichoderma sp. RCK65 and checked for its efficiency in hydrolysis of Prosopis juliflora (a woody substrate). Preliminary studies suggested that when 48 h old secondary fungal culture (20 % v/w) was inoculated in wheat bran moistened with mineral salt solution (pH 4.5 and 1:3 solid to moisture ratio), incubated at 30 °C and after 72 h, it produced maximum cellulase (CMCase 145 U/gds, FPase 38 U/gds and β-glucosidase 105 U/gds). However, using statistical approach a S:L ratio (1:1) was surprisingly found to be optimum that improved cellulase that is CMCase activity by 6.21 %, FPase activity by 23.68 % and β-glucosidase activity by 37.28 %. The estimated cost of crude enzyme (Rs. 5.311/1000 FPase units) seems to be economically feasible which may be due to high enzyme titre, less cultivation time and low media cost. Moreover, when the crude enzyme was used to saccharify pretreated Prosopis juliflora (a woody substrate), it resulted up to 83 % (w/w) saccharification.

Bioethanol production from the dry powder of Jerusalem artichoke tubers by recombinant Saccharomyces cerevisiae in simultaneous saccharification and fermentation.

Science.gov (United States)

Wang, Yi-Zhou; Zou, Shan-Mei; He, Mei-Lin; Wang, Chang-Hai

2015-04-01

It has been found that recombinant Saccharomyces cerevisiae 6525 can produce high concentration of ethanol in one-step fermentation from the extract of Jerusalem artichoke tubers or inulin. However, the utilization rate of raw materials was low and the fermentation process was costly and complicated. Therefore, in this study, after the optimum processing conditions for ethanol production in fed-batch fermentation were determined in flask, the recombinant S. cerevisiae 6525 was first used to produce ethanol from the dry powder of Jerusalem artichoke tubers in 5-L agitating fermentor. After 72 h of fermentation, around 84.3 g/L ethanol was produced in the fermentation liquids, and the conversion efficiency of inulin-type sugars to ethanol was 0.453, or 88.6 % of the theoretical value of 0.511. This study showed high feasibility of bioethanol industrial production from the Jerusalem artichoke tubers and provided a basis for it in the future.

The production of chemicals from food processing wastes using a novel fermenter separator. Annual progress report, January 1993--March 1994

Energy Technology Data Exchange (ETDEWEB)

Dale, M.C.; Venkatesh, K.V.; Choi, H.; Salicetti-Piazza, L.; Borgos-Rubio, N.; Okos, M.R.; Wankat, P.C.

1994-03-15

The basic objective of this project is to convert waste streams from the food processing industry to usable fuels and chemicals using novel bioreactors. These bioreactors should allow economical utilization of waste (whey, waste sugars, waste starch, bottling wastes, candy wastes, molasses, and cellulosic wastes) by the production of ethanol, acetone/butanol, organic acids (acetic, lactic, and gluconic), yeast diacetyl flavor, and antifungal compounds. Continuous processes incorporating various processing improvements such as simultaneous product separation and immobilized cells are being developed to allow commercial scale utilization of waste stream. The production of ethanol by a continuous reactor-separator is the process closest to commercialization with a 7,500 liter pilot plant presently sited at an Iowa site to convert whey lactose to ethanol. Accomplishments during 1993 include installation and start-up of a 7,500 liter ICRS for ethanol production at an industry site in Iowa; Donation and installation of a 200 liter yeast pilot Plant to the project from Kenyon Enterprises; Modeling and testing of a low energy system for recovery of ethanol from vapor is using a solvent absorption/extractive distillation system; Simultaneous saccharification/fermentation of raw corn grits and starch in a stirred reactor/separator; Testing of the ability of `koji` process to ferment raw corn grits in a `no-cook` process.

Production of bio-sugar and bioethanol from coffee residue (CR) by acid-chlorite pretreatment.

Science.gov (United States)

Kim, Ho Myeong; Choi, Yong-Soo; Lee, Dae-Seok; Kim, Yong-Hwan; Bae, Hyeun-Jong

2017-07-01

Nowadays, coffee residue (CR) after roasting is recognized as one of the most useful resources in the world for producing the biofuel and bio-materials. In this study, we evaluated the potential of bio-sugar and bioethanol production from acid-chlorite treated CR. Notably, CR treated three times with acid-chlorite after organic solvent extraction (OSE-3), showed the high monosaccharide content, and the efficient sugar conversion yield compared to the other pretreatment conditions. The OSE-3 (6% substrate loading, w/v) can produce bio-sugar (0.568g/g OSE-3). Also, simultaneous saccharification and fermentation (SSF) produced ethanol (0.266g/g OSE-3), and showed an ethanol conversion yield of 73.8% after a 72-h reaction period. These results suggest that acid-chlorite pretreatment can improve the bio-sugar and bioethanol production of CR by removing the phenolic and brown compounds. Copyright © 2017 Elsevier Ltd. All rights reserved.

Ethanol production from rape straw by a two-stage pretreatment under mild conditions.

Science.gov (United States)

Romero, Inmaculada; López-Linares, Juan C; Delgado, Yaimé; Cara, Cristóbal; Castro, Eulogio

2015-08-01

The growing interest on rape oil as raw material for biodiesel production has resulted in an increasing availability of rape straw, an agricultural residue that is an attractive renewable source for the production of second-generation bioethanol. Pretreatment is one of the key steps in such a conversion process. In this work, a sequential two-stage pretreatment with dilute sulfuric acid (130 °C, 60 min, 2% w/v H2SO4) followed by H2O2 (1-5% w/v) in alkaline medium (NaOH) at low temperature (60, 90 °C) and at different pretreatment times (30-90 min) was investigated. The first-acid stage allows the solubilisation of hemicellulose fraction into fermentable sugars. The second-alkaline peroxide stage allows the delignification of the solid material whilst the cellulose remaining in rape straw turned highly digestible by cellulases. Simultaneous saccharification and fermentation with 15% (w/v) delignified substrate at 90 °C, 5% H2O2 for 60 min, led to a maximum ethanol production of 53 g/L and a yield of 85% of the theoretical.

Combined Mechanical Destruction and Alkaline Pretreatment of Wheat Straw for Enhanced Enzymatic Saccharification

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Qianqian Wang

2014-09-01

Full Text Available Wheat straw was pretreated by combined mechanical destruction and alkaline pretreatments to enhance enzymatic saccharification. Four strategies were employed to evaluate the potential of wheat straw as a feedstock for fermentable sugar production. The effects of the pretreatments on the substrate morphology, size distribution, chemical composition, and cellulose crystallinity, along with the subsequent enzymatic digestibility, were investigated. Optical microscope images showed that mechanical pretreatment alone resulted in poor fiber defibrillation, wherein samples mostly consisted of rigid fiber bundles, while integrated mechanical destruction and alkaline pretreatment led to relatively good fiber defibrillation. Low temperature NaOH/urea pretreatment can fibrillate the rigid fiber bundles into a relatively loose network and alter the structure of the treated substrate to make cellulose more accessible. The glucan conversion rates were 77% and 95% for integrated mechanical destruction and alkaline pretreatments and mechanical destruction followed by low temperature NaOH/urea and ammonium/urea pretreatments, respectively, after 72 h of enzymatic hydrolysis with enzyme loadings of 10 FPU cellulase per g of oven-dry substrate.

Fermentation Process Modeling with Levenberg-Marquardt Algorithm and Runge-Kutta Method on Ethanol Production by Saccharomyces cerevisiae

Directory of Open Access Journals (Sweden)

Dengfeng Liu

2014-01-01

Full Text Available The core of the Chinese rice wine making is a typical simultaneous saccharification and fermentation (SSF process. In order to control and optimize the SSF process of Chinese rice wine brewing, it is necessary to construct kinetic model and study the influence of temperature on the Chinese rice wine brewing process. An unstructured kinetic model containing 12 kinetics parameters was developed and used to describe the changing of kinetic parameters in Chinese rice wine fermentation at 22, 26, and 30°C. The effects of substrate and product inhibitions were included in the model, and four variable, including biomass, ethanol, sugar and substrate were considered. The R-square values for the model are all above 0.95 revealing that the model prediction values could match experimental data very well. Our model conceivably contributes significantly to the improvement of the industrial process for the production of Chinese rice wine.

Highly efficient production of D-lactic acid from chicory-derived inulin by Lactobacillus bulgaricus.

Science.gov (United States)

Xu, Qianqian; Zang, Ying; Zhou, Jie; Liu, Peng; Li, Xin; Yong, Qiang; Ouyang, Jia

2016-11-01

Inulin is a readily available feedstock for cost-effective production of biochemicals. To date, several studies have explored the production of bioethanol, high-fructose syrup and fructooligosaccharide, but there are no studies regarding the production of D-lactic acid using inulin as a carbon source. In the present study, chicory-derived inulin was used for D-lactic acid biosynthesis by Lactobacillus bulgaricus CGMCC 1.6970. Compared with separate hydrolysis and fermentation processes, simultaneous saccharification and fermentation (SSF) has demonstrated the best performance of D-lactic acid production. Because it prevents fructose inhibition and promotes the complete hydrolysis of inulin, the highest D-lactic acid concentration (123.6 ± 0.9 g/L) with a yield of 97.9 % was obtained from 120 g/L inulin by SSF. Moreover, SSF by L. bulgaricus CGMCC 1.6970 offered another distinct advantage with respect to the higher optical purity of D-lactic acid (>99.9 %) and reduced number of residual sugars. The excellent performance of D-lactic acid production from inulin by SSF represents a high-yield method for D-lactic acid production from non-food grains.

Efficient approach for bioethanol production from red seaweed Gelidium amansii.

Science.gov (United States)

Kim, Ho Myeong; Wi, Seung Gon; Jung, Sera; Song, Younho; Bae, Hyeun-Jong

2015-01-01

Gelidium amansii (GA), a red seaweed species, is a popular source of food and chemicals due to its high galactose and glucose content. In this study, we investigated the potential of bioethanol production from autoclave-treated GA (ATGA). The proposed method involved autoclaving GA for 60min for hydrolysis to glucose. Separate hydrolysis and fermentation processing (SHF) achieved a maximum ethanol concentration of 3.33mg/mL, with a conversion yield of 74.7% after 6h (2% substrate loading, w/v). In contrast, simultaneous saccharification and fermentation (SSF) produced an ethanol concentration of 3.78mg/mL, with an ethanol conversion yield of 84.9% after 12h. We also recorded an ethanol concentration of 25.7mg/mL from SSF processing of 15% (w/v) dry matter from ATGA after 24h. These results indicate that autoclaving can improve the glucose and ethanol conversion yield of GA, and that SSF is superior to SHF for ethanol production. Copyright © 2014 Elsevier Ltd. All rights reserved.

Heterologous production of cellobiose dehydrogenases from the basidiomycete Coprinopsis cinerea and the ascomycete Podospora anserina and their effect on saccharification of wheat straw.

Science.gov (United States)

Turbe-Doan, Annick; Arfi, Yonathan; Record, Eric; Estrada-Alvarado, Isabel; Levasseur, Anthony

2013-06-01

Cellobiose dehydrogenases (CDHs) are extracellular glycosylated haemoflavoenzymes produced by many different wood-degrading and phytopathogenic fungi. Putative cellobiose dehydrogenase genes are recurrently discovered by genome sequencing projects in various phylogenetically distinct fungi. The genomes from the basidiomycete Coprinopsis cinerea and the ascomycete Podospora anserina were screened for candidate cdh genes, and one and three putative gene models were evidenced, respectively. Two putative cdh genes were selected and successfully expressed for the first time in Aspergillus niger. CDH activity was measured for both constructions (CDHcc and CDHpa), and both recombinant CDHs were purified to homogeneity and subsequently characterised. Kinetic constants were determined for several carbohydrates including β-1,4-linked di- and oligosaccharides. Optimal temperature and pH were 60 °C and 5 for CDHcc and 65-70 °C and 6 for CDHpa. Both CDHs showed a broad range of pH stability between 4 and 8. The effect of both CDHs on saccharification of micronized wheat straw by an industrial Trichoderma reesei secretome was determined. The addition of each CDH systematically decreased the release of total reducing sugars, but to different extents and according to the CDH concentration. Analytical methods were carried out to quantify the release of glucose, xylose and gluconic acid. An increase of glucose and xylose was measured at a low CDHcc concentration. At moderated and high CDHcc and CDHpa concentrations, glucose was severely reduced with a concomitant increase of gluconic acid. In conclusion, these results give new insights into the physical and chemical parameters and diversity of basidiomycetous and ascomycetous CDHs. These findings also demonstrated that CDH drastically influenced the saccharification on a natural substrate, and thus, CDH origin, concentration and potential enzymatic partners should be carefully considered in future artificial secretomes for

On energy consumption for size-reduction and yields from subsequent enzymatic saccharification of pretreated lodgepole pine

Science.gov (United States)

W. Zhu; Junyong Zhu; Roland Gleisner; X.J. Pan

2010-01-01

This study investigated the effects of chemical pretreatment and disk-milling conditions on energy consumption for size-reduction and the efficiency of enzymatic cellulose saccharification of a softwood. Lodgepole pine wood chips produced from thinnings of a 100-year-old unmanaged forest were pretreated by hot-water, dilute-acid, and two SPORL processes (Sulfite...

Ethanol production using whole plant biomass of Jerusalem artichoke by Kluyveromyces marxianus CBS1555.

Science.gov (United States)

Kim, Seonghun; Park, Jang Min; Kim, Chul Ho

2013-03-01

Jerusalem artichoke is a low-requirement sugar crop containing cellulose and hemicellulose in the stalk and a high content of inulin in the tuber. However, the lignocellulosic component in Jerusalem artichoke stalk reduces the fermentability of the whole plant for efficient bioethanol production. In this study, Jerusalem artichoke stalk was pretreated sequentially with dilute acid and alkali, and then hydrolyzed enzymatically. During enzymatic hydrolysis, approximately 88 % of the glucan and xylan were converted to glucose and xylose, respectively. Batch and fed-batch simultaneous saccharification and fermentation of both pretreated stalk and tuber by Kluyveromyces marxianus CBS1555 were effectively performed, yielding 29.1 and 70.2 g/L ethanol, respectively. In fed-batch fermentation, ethanol productivity was 0.255 g ethanol per gram of dry Jerusalem artichoke biomass, or 0.361 g ethanol per gram of glucose, with a 0.924 g/L/h ethanol productivity. These results show that combining the tuber and the stalk hydrolysate is a useful strategy for whole biomass utilization in effective bioethanol fermentation from Jerusalem artichoke.

Bioethanol production: an integrated process of low substrate loading hydrolysis-high sugars liquid fermentation and solid state fermentation of enzymatic hydrolysis residue.

Science.gov (United States)

Chu, Qiulu; Li, Xin; Ma, Bin; Xu, Yong; Ouyang, Jia; Zhu, Junjun; Yu, Shiyuan; Yong, Qiang

2012-11-01

An integrated process of enzymatic hydrolysis and fermentation was investigated for high ethanol production. The combination of enzymatic hydrolysis at low substrate loading, liquid fermentation of high sugars concentration and solid state fermentation of enzymatic hydrolysis residue was beneficial for conversion of steam explosion pretreated corn stover to ethanol. The results suggested that low substrate loading hydrolysis caused a high enzymatic hydrolysis yield; the liquid fermentation of about 200g/L glucose by Saccharomyces cerevisiae provided a high ethanol concentration which could significantly decrease cost of the subsequent ethanol distillation. A solid state fermentation of enzymatic hydrolysis residue was combined, which was available to enhance ethanol production and cellulose-to-ethanol conversion. The results of solid state fermentation demonstrated that the solid state fermentation process accompanied by simultaneous saccharification and fermentation. Copyright © 2012 Elsevier Ltd. All rights reserved.

Microbial production of mannitol by Lactobacillus brevis 3-A5 from concentrated extract of Jerusalem artichoke tubers.

Science.gov (United States)

Cao, Hailong; Yue, Min; Liu, Gang; Du, Yuguang; Yin, Heng

2018-05-01

In the present study, the conversion of the extract of Jerusalem artichoke tubers for mannitol production by Lactobacillus brevis 3-A5 was investigated. When the bacterium utilized enzymatic hydrolysates of Jerusalem artichoke extract as the main substrates in batch fermentation, the significant decrease in mannitol productivity was observed when the initial concentration of reducing sugar increased. Then, a strategy of continuous fed-batch fermentation was adopted for improving mannitol production with enzymatic hydrolysates of Jerusalem artichoke extract as main substrates. Although the concentration of mannitol could reach 199.86 g/L at the end of the fermentation, the productivity for the overall process of the fermentation was only 1.67 g/L/H. To improve the mannitol productivity with both higher yield and concentration, the simultaneous enzymatic saccharification and fermentation (SSF) was studied. In SSF, the mannitol production reached 176.50 g/L in 28 H with a productivity of 6.30 g/L/H and a yield of 0.68 g/g total sugar. Our study provides a cost-effective and eco-friendly method for mannitol production from a cheap biomass. © 2017 International Union of Biochemistry and Molecular Biology, Inc.

Increased saccharification yields from aspen biomass upon treatment with enzymatically generated peracetic acid.

Science.gov (United States)

Duncan, Shona; Jing, Qing; Katona, Adrian; Kazlauskas, Romas J; Schilling, Jonathan; Tschirner, Ulrike; Aldajani, Waleed Wafa

2010-03-01

The recalcitrance of lignocellulosic biomass to enzymatic release of sugars (saccharification) currently limits its use as feedstock for biofuels. Enzymatic hydrolysis of untreated aspen wood releases only 21.8% of the available sugars due primarily to the lignin barrier. Nature uses oxidative enzymes to selectively degrade lignin in lignocellulosic biomass, but thus far, natural enzymes have been too slow for industrial use. In this study, oxidative pretreatment with commercial peracetic acid (470 mM) removed 40% of the lignin (from 19.9 to 12.0 wt.% lignin) from aspen and enhanced the sugar yields in subsequent enzymatic hydrolysis to about 90%. Increasing the amount of lignin removed correlated with increasing yields of sugar release. Unfortunately, peracetic acid is expensive, and concentrated forms can be hazardous. To reduce costs and hazards associated with using commercial peracetic acid, we used a hydrolase to catalyze the perhydrolysis of ethyl acetate generating 60-70 mM peracetic acid in situ as a pretreatment to remove lignin from aspen wood. A single pretreatment was insufficient, but multiple cycles (up to eight) removed up to 61.7% of the lignin enabling release of >90% of the sugars during saccharification. This value corresponds to a predicted 581 g of fermentable sugars from 1 kg of aspen wood. Improvements in the enzyme stability are needed before the enzymatically generated peracetic acid is a commercially viable alternative.

Cellulase production by Penicillium funiculosum and its application in the hydrolysis of sugar cane bagasse for second generation ethanol production by fed batch operation.

Science.gov (United States)

Maeda, Roberto Nobuyuki; Barcelos, Carolina Araújo; Santa Anna, Lídia Maria Melo; Pereira, Nei

2013-01-10

This study aimed to produce a cellulase blend and to evaluate its application in a simultaneous saccharification and fermentation (SSF) process for second generation ethanol production from sugar cane bagasse. The sugar cane bagasse was subjected to pretreatments (diluted acid and alkaline), as for disorganizing the ligocellulosic complex, and making the cellulose component more amenable to enzymatic hydrolysis. The residual solid fraction was named sugar cane bagasse partially delignified cellulignin (PDC), and was used for enzyme production and ethanol fermentation. The enzyme production was performed in a bioreactor with two inoculum concentrations (5 and 10% v/v). The fermentation inoculated with higher inoculum size reduced the time for maximum enzyme production (from 72 to 48). The enzyme extract was concentrated using tangential ultrafiltration in hollow fiber membranes, and the produced cellulase blend was evaluated for its stability at 37 °C, operation temperature of the simultaneous SSF process, and at 50 °C, optimum temperature of cellulase blend activity. The cellulolytic preparation was stable for at least 300 h at both 37 °C and 50 °C. The ethanol production was carried out by PDC fed-batch SSF process, using the onsite cellulase blend. The feeding strategy circumvented the classic problems of diffusion limitations by diminishing the presence of a high solid:liquid ratio at any time, resulting in high ethanol concentration at the end of the process (100 g/L), which corresponded to a fermentation efficiency of 78% of the maximum obtainable theoretically. The experimental results led to the ratio of 380 L of ethanol per ton of sugar cane bagasse PDC. Copyright © 2012 Elsevier B.V. All rights reserved.

Different Routes for Conifer- and Sinapaldehyde and Higher Saccharification upon Deficiency in the Dehydrogenase CAD11[OPEN

Science.gov (United States)

Laurans, Françoise; Foster, Cliff; Légée, Frédéric

2017-01-01

In the search for renewable energy sources, genetic engineering is a promising strategy to improve plant cell wall composition for biofuel and bioproducts generation. Lignin is a major factor determining saccharification efficiency and, therefore, is a prime target to engineer. Here, lignin content and composition were modified in poplar (Populus tremula × Populus alba) by specifically down-regulating CINNAMYL ALCOHOL DEHYDROGENASE1 (CAD1) by a hairpin-RNA-mediated silencing approach, which resulted in only 5% residual CAD1 transcript abundance. These transgenic lines showed no biomass penalty despite a 10% reduction in Klason lignin content and severe shifts in lignin composition. Nuclear magnetic resonance spectroscopy and thioacidolysis revealed a strong increase (up to 20-fold) in sinapaldehyde incorporation into lignin, whereas coniferaldehyde was not increased markedly. Accordingly, ultra-high-performance liquid chromatography-mass spectrometry-based phenolic profiling revealed a more than 24,000-fold accumulation of a newly identified compound made from 8-8 coupling of two sinapaldehyde radicals. However, no additional cinnamaldehyde coupling products could be detected in the CAD1-deficient poplars. Instead, the transgenic lines accumulated a range of hydroxycinnamate-derived metabolites, of which the most prominent accumulation (over 8,500-fold) was observed for a compound that was identified by purification and nuclear magnetic resonance as syringyl lactic acid hexoside. Our data suggest that, upon down-regulation of CAD1, coniferaldehyde is converted into ferulic acid and derivatives, whereas sinapaldehyde is either oxidatively coupled into S′(8-8)S′ and lignin or converted to sinapic acid and derivatives. The most prominent sink of the increased flux to hydroxycinnamates is syringyl lactic acid hexoside. Furthermore, low-extent saccharification assays, under different pretreatment conditions, showed strongly increased glucose (up to +81%) and xylose

Bioethanol production from ball milled bagasse using an on-site produced fungal enzyme cocktail and xylose-fermenting Pichia stipitis.

Science.gov (United States)

Buaban, Benchaporn; Inoue, Hiroyuki; Yano, Shinichi; Tanapongpipat, Sutipa; Ruanglek, Vasimon; Champreda, Verawat; Pichyangkura, Rath; Rengpipat, Sirirat; Eurwilaichitr, Lily

2010-07-01

Sugarcane bagasse is one of the most promising agricultural by-products for conversion to biofuels. Here, ethanol fermentation from bagasse has been achieved using an integrated process combining mechanical pretreatment by ball milling, with enzymatic hydrolysis and fermentation. Ball milling for 2 h was sufficient for nearly complete cellulose structural transformation to an accessible amorphous form. The pretreated cellulosic residues were hydrolyzed by a crude enzyme preparation from Penicillium chrysogenum BCC4504 containing cellulase activity combined with Aspergillus flavus BCC7179 preparation containing complementary beta-glucosidase activity. Saccharification yields of 84.0% and 70.4% for glucose and xylose, respectively, were obtained after hydrolysis at 45 degrees C, pH 5 for 72 h, which were slightly higher than those obtained with a commercial enzyme mixture containing Acremonium cellulase and Optimash BG. A high conversion yield of undetoxified pretreated bagasse (5%, w/v) hydrolysate to ethanol was attained by separate hydrolysis and fermentation processes using Pichia stipitis BCC15191, at pH 5.5, 30 degrees C for 24 h resulting in an ethanol concentration of 8.4 g/l, corresponding to a conversion yield of 0.29 g ethanol/g available fermentable sugars. Comparable ethanol conversion efficiency was obtained by a simultaneous saccharification and fermentation process which led to production of 8.0 g/l ethanol after 72 h fermentation under the same conditions. This study thus demonstrated the potential use of a simple integrated process with minimal environmental impact with the use of promising alternative on-site enzymes and yeast for the production of ethanol from this potent lignocellulosic biomass. 2009. Published by Elsevier B.V.

Pretreatment and Fractionation of Wheat Straw for Production of Fuel Ethanol and Value-added Co-products in a Biorefinery

Directory of Open Access Journals (Sweden)

Xiu Zhang

2014-08-01

Full Text Available An integrated process has been developed for a wheat straw biorefinery. In this process, wheat straw was pretreated by soaking in aqueous ammonia (SAA, which extensively removed lignin but preserved high percentages of the carbohydrate fractions for subsequent bioconversion. The pretreatment conditions included 15 wt% NH4OH, 1:10 solid:liquid ratio, 65 oC and 15 hours. Under these conditions, 48% of the original lignin was removed, whereas 98%, 83% and 78% of the original glucan, xylan, and arabinan, respectively, were preserved. The pretreated material was subsequently hydrolyzed with a commercial hemicellulase to produce a solution rich in xylose and low in glucose plus a cellulose-enriched solid residue. The xylose-rich solution then was used for production of value-added products. Xylitol and astaxanthin were selected to demonstrate the fermentability of the xylose-rich hydrolysate. Candida mogii and Phaffia rhodozyma were used for xylitol and astaxanthin fermentation, respectively. The cellulose-enriched residue obtained after the enzymatic hydrolysis of the pretreated straw was used for ethanol production in a fed-batch simultaneous saccharification and fermentation (SSF process. In this process, a commercial cellulase was used for hydrolysis of the glucan in the residue and Saccharomyces cerevisiae, which is the most efficient commercial ethanol-producing organism, was used for ethanol production. Final ethanol concentration of 57 g/l was obtained at 27 wt% total solid loading.

Pretreatment of wheat straw by nonionic surfactant-assisted dilute acid for enhancing enzymatic hydrolysis and ethanol production.

Science.gov (United States)

Qi, Benkun; Chen, Xiangrong; Wan, Yinhua

2010-07-01

Pretreating wheat straw (WS) with combined use of varied sulfuric acid concentration (0-3%, w/v) and Tween 20 concentration (0-1%) was investigated in an attempt to enhance the hydrolysis and fermentability of pretreated WS. Enzymatic hydrolysis yield of glucan and xylan and ethanol production by simultaneous saccharification and fermentation (SSF) of water-insoluble solids (WIS) were significantly affected by the amount of Tween 20 added during acid pretreatment. Any further addition of Tween 20 in either hydrolysis stage or fermentation stage only led to small increase in glucan conversion and ethanol production. Determination of adsorption of cellulases during hydrolysis showed that Tween 20-assisted acid treated straw solution contained more free cellulases than individual acid treated straw solution, indicating that modification of lignin surface by Tween 20 added during pretreatment likely occurred. In addition, the effects of pretreatment conditions on overall recovery of glucose and xylose after pretreatment and enzymatic hydrolysis were also investigated. Copyright 2010 Elsevier Ltd. All rights reserved.

Monosaccharides and Ethanol Production from Superfine Ground Sugarcane Bagasse Using Enzyme Cocktail

Directory of Open Access Journals (Sweden)

Jingbo Li

2014-03-01

Full Text Available In this work, the effect of particle size on the enzymatic hydrolysis of milled and sieved sugarcane bagasse (SCB was studied. The enzymatic hydrolysis and fermentability of superfine ground SCB (SGP400 using an enzyme cocktail strategy were also explored. Particle size reduction improved the enzymatic hydrolysis. The highest glucose yield was 44.75%, which was obtained from SGP400. The enzyme cocktail strategy greatly enhanced the glucose and xylose yield. The maximum glucose and xylose yield was from the enzyme cocktail of cellulase, xylanase, and pectinase. Synergistic action between xylanase and pectinase as well as cellulase and pectinase was quite noticeable. Hydrolysis times affected the degree of synergism. Ethanol production was carried out by employing simultaneous saccharification and fermentation (SSF and semi-SSF using enzymes and their cocktails. Semi-SSF was found to be the better one compared with SSF. Xylanase and pectinase aided the ethanol production in both fermentation modes. Ethanol yield was 7.81 and 7.30 g/L for semi-SSF and SSF, respectively by using an enzyme cocktail of cellulase, β-glucosidase, pectinase, and xylanase.

High-solids loading enzymatic hydrolysis of waste papers for biofuel production

International Nuclear Information System (INIS)

Wang, Lei; Templer, Richard; Murphy, Richard J.

2012-01-01

Highlights: ► Waste papers have great potential as a feedstock for bioethanol production. ► A wet blending step would significantly enhance enzymatic hydrolysis efficiency. ► High-solids loading saccharification was performed successfully on waste papers. ► Saccharification data were from four types of paper and two enzyme alternatives. ► Enzymatic hydrolysis kinetic models were validated by experimental data. -- Abstract: Waste papers (newspaper, office paper, magazines and cardboard in this study) with 50–73% (w/w oven dry weight) carbohydrate contents have considerable potential as raw materials for bioethanol production. A particle size reduction step of wet blending prior to enzymatic hydrolysis of newspaper was found to increase the glucan conversion efficiency by up to 10%. High-solids loading hydrolysis at 15% (w/w) of four types of paper using two enzyme alternatives, Celluclast 1.5L supplemented with Novozyme 188 and Cellic Ctec 1 (Novozymes A/S, Demark), at various enzyme concentrations were successfully performed in a lab-scale overhead-stirred reactor. This work has identified the relative saccharification performance for the four types of paper and shows office paper and cardboard to be more suitable for producing bioethanol than newspaper or magazine paper. The experimental data were also very well described by a modified, simple three parameter glucan and xylan hydrolysis model. These findings provide the possibility for incorporating this validated kinetic model into process designs required for commercial scale bioethanol production from waste paper resources.

Facilitating the enzymatic saccharification of pulped bamboo residues by degrading the remained xylan and lignin-carbohydrates complexes.

Science.gov (United States)

Huang, Caoxing; He, Juan; Li, Xin; Min, Douyong; Yong, Qiang

2015-09-01

Kraft pulping was performed on bamboo residues and its impact on the chemical compositions and the enzymatic digestibility of the samples were investigated. To improve the digestibility of sample by degrading the xylan and lignin-carbohydrates complexes (LCCs), xylanase and α-L-arabinofuranosidase (AF) were supplemented with cellulase. The results showed more carbohydrates were remained in the samples pulped with low effective alkali (EA) charge, compared to conventional kraft pulping. When 120 IU/g xylanase and 15 IU/g AF were supplemented with 20 FPU/g cellulase, the xylan degradation yield of the sample pulped with 12% EA charge increased from 68.20% to 88.35%, resulting in an increased enzymatic saccharification efficiency from 58.98% to 83.23%. The amount of LCCs in this sample decreased from 8.63/100C9 to 2.99/100C9 after saccharification with these enzymes. The results indicated that degrading the remained xylan and LCCs in the pulp could improve its enzymatic digestibility. Copyright © 2015 Elsevier Ltd. All rights reserved.

pH-Induced Lignin Surface Modification to Reduce Nonspecific Cellulase Binding and Enhance Enzymatic Saccharification of Lignocelluloses

Science.gov (United States)

Hongming Lou; J.Y. Zhu; Tian Qing Lan; Huranran Lai; Xueqing Qiu

2013-01-01

We studied the mechanism of the significant enhancement in the enzymatic saccharification of lignocelluloses at an elevated pH of 5.5–6.0. Four lignin residues with different sulfonic acid contents were isolated from enzymatic hydrolysis of lodgepole pine pretreated by either dilute acid (DA) or sulfite pretreatment to overcome recalcitrance of lignocelluloses (SPORL...

Biomimetic cell wall model studies to identify new lignin bioengineering targets for improving biomass susceptibility to pretreatment and enzymatic saccharification

Science.gov (United States)

Increasingly, bioengineering of lignin to contain atypical building blocks from other metabolic pathways is being pursued to custom-design lignin that is easier to remove by chemical pretreatments and less inhibitory toward polysaccharide saccharification. Because plants produce such a diverse array...

Production of bio ethanol from waste potatoes

Science.gov (United States)

Jaber Noufal, Mohamad; Li, Baizhan; Maalla, Zena Ali

2017-03-01

In this research, production of ethanol from waste potatoes fermentation was studied using Saccharomyces cerevisiae. Potato Flour prepared from potato tubers after cooking and drying at 85°C. A homogenous slurry of potato flour prepared in water at solid-liquid ratio 1:10. Liquefaction of potato starch slurry was done with α-amylase at 80°C for 40 min followed by saccharification process which was done with glucoamylase at 65°C for two hr. Fermentation of hydrolysate with Saccharomyces cerevisiae at 35°C for two days resulted in the production of 33 g/l ethanol. The following parameters have been analysed: temperature, time of fermentation and pH. It found that Saccharification process is affected by enzyme Amylase 300 concentration and concentration of 1000μl/100ml gives the efficient effect of the process. The best temperature for fermentation process was found to be about 35°C. Also, it noticed that ethanol production increased as a time of fermentation increased but after 48 hr further growth in fermentation time did not have an appreciable effect. Finally, the optimal value of pH for fermentation process was about 5 to 6.

Production of high concentration of l-lactic acid from oil palm empty fruit bunch by thermophilic Bacillus coagulans JI12.

Science.gov (United States)

Juturu, Veeresh; Wu, Jin Chuan

2018-03-01

Thermophilic Bacillus coagulans JI12 was used to ferment hemicellulose hydrolysate obtained by acid hydrolysis of oil palm empty fruit bunch at 50 °C and pH 6, producing 105.4 g/L of l-lactic acid with a productivity of 9.3 g/L/H by fed-batch fermentation under unsterilized conditions. Simultaneous saccharification and fermentation (SSF) was performed at pH 5.5 and 50 °C to convert both hemicellulose hydrolysate and cellulose-lignin complex in the presence of Cellic Ctec2 cellulases using yeast extract (20 g/L) as the nitrogen source, giving 114.0 g/L of l-lactic acid with a productivity of 5.7 g/L/H. The SSF was also conducted by replacing yeast extract with equal amount of dry Bakers' yeast, achieving 120.0 g/L of l-lactic acid with a productivity of 4.3 g/L/H. To the best of our knowledge, these lactic acid titers and productivities are the highest ever reported from lignocellulose hydrolysates. © 2017 International Union of Biochemistry and Molecular Biology, Inc.

Relationship to reducing sugar production and scanning electron microscope structure to pretreated hemp hurd biomass (Cannabis sativa)

International Nuclear Information System (INIS)

Abraham, Reinu E.; Barrow, Colin J.; Puri, Munish

2013-01-01

Lignocellulosic biomass is a highly rigid and recalcitrant structure which requires pretreatment to loosen chemical bonds to make accessible monomeric sugars for biofuel production. In this study, locally available biomass, that is hemp (Cannabis sativa), a low cost feedstock for ethanol production, has been used for the production of fermentable sugars. Hemp hurd biomass (HHB) was exposed to five different pretreatments which included dilute acid (H 2 SO 4 ), alkaline (NaOH), alkaline peroxide, hot water and one stage dilute acid (H 2 SO 4 ). Different pretreatments resulted in loosening and degradation of HHB structure thus facilitating enzymatic saccharification at optimized parameters (pH–4.8 and 50 °C). The changes in the reactive groups (hydroxyl or acetyl) of the HHB were confirmed by attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy. Scanning electron microscopy (SEM) was employed to characterize the surface morphology of untreated and treated HHB. Finally, enzymatic saccharification demonstrated maximum yield of total sugars (743 mg g −1 ) that are suitable for biofuel production. -- Highlights: • Hemp hurd biomass (HHB) was used for producing fermentable sugars. • Alkaline pretreatment resulted in loosening and degradation of hemp structure. • Pretreated HHB was characterized using FTIR studies. • SEM studies evaluated the opening of fiber bundles in pretreatment, thereby increasing cellulose access to enzymes. • Enzymatic saccharification of pretreated HHB demonstrated maximum yield of reducing sugars

Pretreatment of the macroalgae Chaetomorpha linum for the production of bioethanol - Comparison of five pretreatment technologies

DEFF Research Database (Denmark)

Schultz-Jensen, Nadja; Thygesen, Anders; Thomsen, Sune Tjalfe

2013-01-01

-assisted pretreatment (PAP) and ball milling (BM), to determine effects of the pretreatment methods on the conversion of C. linum into ethanol by simultaneous saccharification and fermentation (SSF). WO and BM showed the highest ethanol yield of 44. g ethanol/100. g glucan, which was close to the theoretical ethanol......A qualified estimate for pretreatment of the macroalgae Chaetomorpha linum for ethanol production was given, based on the experience of pretreatment of land-based biomass. C. linum was subjected to hydrothermal pretreatment (HTT), wet oxidation (WO), steam explosion (STEX), plasma...... yield of 57. g ethanol/100. g glucan. A 64% higher ethanol yield, based on raw material, was reached after pretreatment with WO and BM compared with unpretreated C. linum, however 50% of the biomass was lost during WO. Results indicated that the right combination of pretreatment and marine macroalgae...

Green coconut mesocarp pretreated by an alkaline process as raw material for bioethanol production.

Science.gov (United States)

Soares, Jimmy; Demeke, Mekonnen M; Foulquié-Moreno, Maria R; Van de Velde, Miet; Verplaetse, Alex; Fernandes, Antonio Alberto Ribeiro; Thevelein, Johan M; Fernandes, Patricia Machado Bueno

2016-09-01

Cocos nucifera L., coconut, is a palm of high importance in the food industry, but a considerable part of the biomass is inedible. In this study, the pretreatment and saccharification parameters NaOH solution, pretreatment duration and enzyme load were evaluated for the production of hydrolysates from green coconut mesocarp using 18% (w/v) total solids (TS). Hydrolysates were not detoxified in order to preserve sugars solubilized during the pretreatment. Reduction of enzyme load from 15 to 7.5 filter paper cellulase unit (FPU)/g of biomass has little effect on the final ethanol titer. With optimized pretreatment and saccharification, hydrolysates with more than 7% (w/v) sugars were produced in 48h. Fermentation of the hydrolysate using industrial Saccharomyces cerevisiae strains produced 3.73% (v/v) ethanol. Our results showed a simple pretreatment condition with a high-solid load of biomass followed by saccharification and fermentation of undetoxified coconut mesocarp hydrolysates to produce ethanol with high titer. Copyright © 2016 Elsevier Ltd. All rights reserved.

The effect of delignification process with alkaline peroxide on lactic acid production from furfural residues

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Yong Tang

2012-11-01

Full Text Available Furfural residues produced from the furfural industry were investigated as a substrate for lactic acid production by simultaneous saccharification and fermentation (SSF. Alkaline peroxide was used for delignification of furfural residues to improve the final lactic acid concentration. The residue was treated with 1.3% to 1.7% hydrogen peroxide at 80 °C for 1 h with a substrate concentration of 3.33%. SSF of furfural residues with different delignification degrees were carried out to evaluate the effect of delignification degree on lactic acid production. Using corn hydrolysates/ furfural residues as substrates, SSF with different media were carried out to investigate the effect of lignin on the interaction between enzymes and lactic acid bacteria. Lactic acid bacteria had a negative effect on cellulase, thus resulting in the reduction of enzyme activity. Lignin and nutrients slowed down the decreasing trend of enzyme activity. A higher delignification resulted in a slower fermentation rate and lower yield due to degradation products of lignin and the effect of lignin on the interaction between enzymes and lactic acid bacteria. For the purpose of lactic acid production, a moderate delignification (furfural residues with the lignin content of 14.8% was optimum.

Different Routes for Conifer- and Sinapaldehyde and Higher Saccharification upon Deficiency in the Dehydrogenase CAD1.

Science.gov (United States)

Van Acker, Rebecca; Déjardin, Annabelle; Desmet, Sandrien; Hoengenaert, Lennart; Vanholme, Ruben; Morreel, Kris; Laurans, Françoise; Kim, Hoon; Santoro, Nicholas; Foster, Cliff; Goeminne, Geert; Légée, Frédéric; Lapierre, Catherine; Pilate, Gilles; Ralph, John; Boerjan, Wout

2017-11-01

In the search for renewable energy sources, genetic engineering is a promising strategy to improve plant cell wall composition for biofuel and bioproducts generation. Lignin is a major factor determining saccharification efficiency and, therefore, is a prime target to engineer. Here, lignin content and composition were modified in poplar ( Populus tremula × Populus alba ) by specifically down-regulating CINNAMYL ALCOHOL DEHYDROGENASE1 ( CAD1 ) by a hairpin-RNA-mediated silencing approach, which resulted in only 5% residual CAD1 transcript abundance. These transgenic lines showed no biomass penalty despite a 10% reduction in Klason lignin content and severe shifts in lignin composition. Nuclear magnetic resonance spectroscopy and thioacidolysis revealed a strong increase (up to 20-fold) in sinapaldehyde incorporation into lignin, whereas coniferaldehyde was not increased markedly. Accordingly, ultra-high-performance liquid chromatography-mass spectrometry-based phenolic profiling revealed a more than 24,000-fold accumulation of a newly identified compound made from 8-8 coupling of two sinapaldehyde radicals. However, no additional cinnamaldehyde coupling products could be detected in the CAD1-deficient poplars. Instead, the transgenic lines accumulated a range of hydroxycinnamate-derived metabolites, of which the most prominent accumulation (over 8,500-fold) was observed for a compound that was identified by purification and nuclear magnetic resonance as syringyl lactic acid hexoside. Our data suggest that, upon down-regulation of CAD1 , coniferaldehyde is converted into ferulic acid and derivatives, whereas sinapaldehyde is either oxidatively coupled into S'(8-8)S' and lignin or converted to sinapic acid and derivatives. The most prominent sink of the increased flux to hydroxycinnamates is syringyl lactic acid hexoside. Furthermore, low-extent saccharification assays, under different pretreatment conditions, showed strongly increased glucose (up to +81%) and

Steam pretreatment of dry and ensiled industrial hemp for ethanol production

Energy Technology Data Exchange (ETDEWEB)

Sipos, Balint; Reczey, Kati [Budapest University of Technology and Economics, Department of Applied Biotechnology and Food Science, Szt. Gellert ter 4., H-1111 Budapest (Hungary); Kreuger, Emma; Bjoernsson, Lovisa [Lund University, Department of Biotechnology, P.O. Box 124, SE-221 00 Lund (Sweden); Svensson, Sven-Erik [Swedish University of Agricultural Sciences, Department of Agriculture - Farming Systems, Technology and Product Quality, P.O. Box 104, SE-230 53 Alnarp (Sweden); Zacchi, Guido [Lund University, Department of Chemical Engineering, P.O. Box 124, SE-221 00 Lund (Sweden)

2010-12-15

Biomass can be converted into liquid and gaseous biofuels with good efficiency. In this study, the conversion of industrial hemp (Cannabis sativa L.), a biomass source that can be cultivated with a high biomass yield per hectare, was used. Steam pretreatment of dry and ensiled hemp was investigated prior to ethanol production. The pretreatment efficiency was evaluated in terms of sugar recovery and polysaccharide conversion in the enzymatic hydrolysis step. For both materials, impregnation with 2% SO{sub 2} followed by steam pretreatment at 210 C for 5 min were found to be the optimal conditions leading to the highest overall yield of glucose. Simultaneous saccharification and fermentation experiments carried out with optimised pretreatment conditions resulted in ethanol yields of 163 g kg{sup -1} ensiled hemp (dry matter) (71% of the theoretical maximum) and 171 g kg{sup -1} dry hemp (74%), which corresponds to 206-216 l Mg{sup -1} ethanol based on initial dry material. (author)

Green Biorefinery of Giant Miscanthus for Growing Microalgae and Biofuel Production

Directory of Open Access Journals (Sweden)

Shuangning Xiu

2017-12-01

Full Text Available In this study, an innovative green biorefinery system was successfully developed to process the green biomass into multiple biofuels and bioproducts. In particular, fresh giant miscanthus was separated into a solid stream (press cake and a liquid stream (press juice using a screw press. The juice was used to cultivate microalga Chlorella vulgaris, which was further thermochemically converted via thermogravimetry analysis (TGA and pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS analysis, resulting in an approximately 80% conversion. In addition, the solid cake of miscanthus was pretreated with dilute sulfuric acid and used as the feedstock for bioethanol production. The results showed that the miscanthus juice could be a highly nutritious source for microalgae that are a promising feedstock for biofuels. The highest cell density was observed in the 15% juice medium. Sugars released from the miscanthus cake were efficiently fermented to ethanol using Saccharomyces cerevisiae through a simultaneous saccharification and fermentation (SSF process, with 88.4% of the theoretical yield.

Utilization of microwave and ultrasound pretreatments in the production of bioethanol from corn

Energy Technology Data Exchange (ETDEWEB)

Nikolic, Svetlana; Mojovic, Ljiljana; Rakin, Marica [University of Belgrade, Faculty of Technology and Metallurgy, Belgrade (Serbia); Pejin, Dusanka; Pejin, Jelena [University of Novi Sad, Faculty of Technology, Novi Sad (Serbia)

2011-08-15

Bioethanol production by simultaneous saccharification and fermentation (SSF) of corn meal by Saccharomyces cerevisiae var. ellipsoideus yeast in a batch system with prior ultrasound or microwave treatment was studied. The optimal duration of the pretreatments and the SSF process kinetics were assessed and determined. Also, the effect of ultrasound and microwave pretreatments on ethanol yield and productivity was investigated. An optimal duration of 5 min was determined for both pretreatments. Ultrasonic and microwave pretreatments effectively increased the glucose concentration obtained after liquefaction by 6.82 and 8.48%, respectively, compared to untreated control sample. Also, both pretreatments improved ethanol yield and productivity during the SSF process. Ultrasound and microwave pretreatments increased the maximum ethanol concentration produced in the SSF process by 11.15 and 13.40% (compared to the control sample), respectively. The application of microwave pretreatment resulted in higher glucose release during liquefaction and consequently in higher ethanol concentration, compared to ultrasound pretreatment. A maximum ethanol concentration of 9.91% (w/w) and percentage of theoretical ethanol yield of 92.27% were achieved after 44 h of the SSF process of corn meal with prior microwave treatment. (orig.)

Improved enzymatic saccharification of steam exploded cotton stalk using alkaline extraction and fermentation of cellulosic sugars into ethanol.

Science.gov (United States)

Keshav, Praveen K; Naseeruddin, Shaik; Rao, L Venkateswar

2016-08-01

Cotton stalk, a widely available and cheap agricultural residue lacking economic alternatives, was subjected to steam explosion in the range 170-200°C for 5min. Steam explosion at 200°C and 5min led to significant hemicellulose solubilization (71.90±0.10%). Alkaline extraction of steam exploded cotton stalk (SECOH) using 3% NaOH at room temperature for 6h led to 85.07±1.43% lignin removal with complete hemicellulose solubilization. Besides, this combined pretreatment allowed a high recovery of the cellulosic fraction from the biomass. Enzymatic saccharification was studied between steam exploded cotton stalk (SECS) and SECOH using different cellulase loadings. SECOH gave a maximum of 785.30±8.28mg/g reducing sugars with saccharification efficiency of 82.13±0.72%. Subsequently, fermentation of SECOH hydrolysate containing sugars (68.20±1.16g/L) with Saccharomyces cerevisiae produced 23.17±0.84g/L ethanol with 0.44g/g yield. Copyright © 2016 Elsevier Ltd. All rights reserved.

Different Spectrophotometric Methods for Simultaneous Determination of Trelagliptin and Its Acid Degradation Product

Science.gov (United States)

Hassan, Mostafa A.; Zaghary, Wafaa A.

2018-01-01

New spectrophotometric and chemometric methods were carried out for the simultaneous assay of trelagliptin (TRG) and its acid degradation product (TAD) and applied successfully as a stability indicating assay to recently approved Zafatek® tablets. TAD was monitored using TLC to ensure complete degradation. Furthermore, HPLC was used to confirm dealing with one major acid degradation product. The proposed methods were developed by manipulating zero-order, first-derivative, and ratio spectra of TRG and TAD using simultaneous equation, first-derivative, and mean-centering methods, respectively. Using Spectra Manager II and Minitab v.14 software, the absorbance at 274 nm–260.4 nm, amplitudes at 260.4 nm–274.0 nm, and mean-centered values at 287.6 nm–257.2 nm were measured against methanol as a blank for TRG and TAD, respectively. Linearity and the other validation parameters were acceptable at concentration ranges of 5–50 μg/mL and 2.5–25 μg/mL for TRG and TAD, respectively. Using one-way analysis of variance (ANOVA), the optimized methods were compared and proved to be accurate for the simultaneous assay of TRG and TAD. PMID:29629213

Simultaneous growth and metabolite production by yoghurt starters and probiotics: a metabolomics approach

NARCIS (Netherlands)

Settachaimongkon, S.

2014-01-01

The main objective of this research was to investigate the simultaneous growth and metabolite production by yoghurt starters and different probiotic strains, i.e. Lactobacillus rhamnosus GG, Bifidobacterium animalis subsp. lactis BB12 and Lactobacillus