Improving the performance of enzymes in hydrolysis of high solids paper pulp derived from MSW

Biotechnology for Biofuels

Dhivya J Puri¹  Sonia Heaven¹  Charles J Banks¹ 

[1] Faculty of Engineering and the Environment, University of Southampton, Southampton, SO17 1BJ, United Kingdom

关键词: Bioethanol;    Municipal solid waste (MSW);    High solids hydrolysis;    Enzyme hydrolysis;    Cellulases;    Lignocellulose;   

Others  :  797965
DOI  :  10.1186/1754-6834-6-107

 received in 2013-05-10, accepted in 2013-07-23,  发布年份 2013

【 摘 要 】

Background

The research aimed to improve the overall conversion efficiency of the CTec® family of enzymes by identifying factors that lead to inhibition and seeking methods to overcome these through process modification and manipulation. The starting material was pulp derived from municipal solid waste and processed in an industrial-scale washing plant.

Results

Analysis of the pulp by acid hydrolysis showed a ratio of 55 : 12 : 6 : 24 : 3 of glucan : xylan : araban/galactan/mannan : lignin : ash. At high total solids content (>18.5% TS) single-stage enzyme hydrolysis gave a maximum glucan conversion of 68%. It was found that two-stage hydrolysis could give higher conversion if sugar inhibition was removed by an intermediate fermentation step between hydrolysis stages. This, however, was not as effective as direct removal of the sugar products, including xylose, by washing of the residual pulp at pH 5. This improved the water availability and allowed reactivation of the pulp-bound enzymes. Inhibition of enzyme activity could further be alleviated by replenishment of β-glucosidase which was shown to be removed during the wash step.

Conclusions

The two-stage hydrolysis process developed could give an overall glucan conversion of 88%, with an average glucose concentration close to 8% in 4 days, thus providing an ideal starting point for ethanol fermentation with a likely yield of 4 wt%. This is a significant improvement over a single-step process. This hydrolysis configuration also provides the potential to recover the sugars associated with residual solids which are diluted when washing hydrolysed pulp.

【 授权许可】

   
2013 Puri et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Licht FO: World Ethanol Markets: The outlook to 2015. UK: Tunbridge Wells; 2006.
• [2]Larsson M, Zacchi G: Production of ethanol from dilute glucose solutions A technical-economic evaluation of various refining alternatives. Bioprocess Biosyst Eng 1996, 15:125-132.
• [3]Larsen J, Østergaard Petersen M, Thirup L, Wen Li H, Krogh Iversen F: The IBUS Process – Lignocellulosic Bioethanol Close to a Commercial Reality. Chem Eng Technol 2008, 31:765-772.
• [4]Wang W, Kang L, Wei H, Arora R, Lee Y: Study on the decreased sugar yield in enzymatic hydrolysis of cellulosic substrate at high solid loading. Appl Biochem Biotechnol 2011, 164:1139-1149.
• [5]Kristensen JB, Felby C, Jorgensen H: Yield-determining factors in high-solids enzymatic hydrolysis of lignocellulose. Biotechnology for Biofuels 2009, 2:10. BioMed Central Full Text
• [6]Yu ZY, Jameel H, Chang HM, Philips R, Park S: Evaluation of the factors affecting avicel reactivity using multi-stage enzymatic hydrolysis. Biotechnol Bioeng 2012, 109:1131-1139.
• [7]Roche C, Dibble C, Stickel J: Laboratory-scale method for enzymatic saccharification of lignocellulosic biomass at high-solids loadings. Biotechnology for Biofuels 2009, 2:28. BioMed Central Full Text
• [8]Xiao ZZ, Zhang X, Gregg DJ, Saddler JN: Effects of sugar inhibition on cellulases and beta-glucosidase during enzymatic hydrolysis of softwood substrates. Appl Biochem Biotechnol 2004, 113:1115-1126.
• [9]Yang J, Zhang X, Yong Q, Yu S: Three-stage hydrolysis to enhance enzymatic saccharification of steam-exploded corn stover. Bioresour Technol 2010, 101:4930-4935.
• [10]Roberts KM, Lavenson DM, Tozzi EJ, McCarthy MJ, Jeoh T: The effects of water interactions in cellulose suspensions on mass transfer and saccharification efficiency at high solids loadings. Cellulose 2011, 18:759-773.
• [11]Selig MJ, Hsieh C-WC, Thygesen LG, Himmel ME, Felby C, Decker SR: Considering water availability and the effect of solute concentration on high solids saccharification of lignocellulosic biomass. Biotechnol Prog 2012, 28:1478-1490.
• [12]Rahikainen J, Mikander S, Marjamaa K, Tamminen T, Lappas A, Viikari L, Kruus K: Inhibition of enzymatic hydrolysis by residual lignins from softwood-study of enzyme binding and inactivation on lignin-rich surface. Biotechnol Bioeng 2011, 108:2823-2834.
• [13]Yang MH, Li WL, Liu BB, Li Q, Xing JM: High-concentration sugars production from corn stover based on combined pretreatments and fed-batch process. Bioresour Technol 2010, 101:4884-4888.
• [14]Andric P, Meyer AS, Jensen PA, Dam-Johansen K: Reactor design for minimizing product inhibition during enzymatic lignocellulose hydrolysis: I. Significance and mechanism of cellobiose and glucose inhibition on cellulolytic enzymes. Biotechnol Adv 2010, 28:308-324.
• [15]Olofsson K, Bertilsson M, Liden G: A short review on SSF - an interesting process option for ethanol production from lignocellulosic feedstocks. Biotechnology for Biofuels 2008, 1(1):1-14. BioMed Central Full Text
• [16]Kemppainen K, Ranta L, Sipilä E, Östman A, Vehmaanperä J, Puranen T, Langfelder K, Hannula J, Kallioinen A, Siika-aho M, et al.: Ethanol and biogas production from waste fibre and fibre sludge – the FibreEtOH concept. Biomass and Bioenergy 2012, 46:60-69.
• [17]Watanabe I, Miyata N, Ando A, Shiroma R, Tokuyasu K, Nakamura T: Ethanol production by repeated-batch simultaneous saccharification and fermentation (SSF) of alkali-treated rice straw using immobilized Saccharomyces cerevisiae cells. Bioresour Technol 2012, 123:695-698.
• [18]Viikari L, Vehmaanperä J, Koivula A: Lignocellulosic ethanol: from science to industry. Biomass and Bioenergy 2012, 46:13-24.
• [19]Holtzapple M, Cognata M, Shu Y, Hendrickson C: Inhibition of Trichoderma reesei cellulase by sugars and solvents. Biotechnol Bioeng 1990, 36:275-287.
• [20]Wu Z, Lee YY: Inhibition of the enzymatic hydrolysis of cellulose by ethanol. Biotechnol Lett 1997, 19:977-979.
• [21]Mores W, Knutsen J, Davis R: Cellulase recovery via membrane filtration. Appl Biochem Biotechnol 2001, 91–93:297-309.
• [22]Zhang MJ, Su RX, Li QA, Qi W, He ZM: Enzymatic saccharification of pretreated corn stover in a fed-batch membrane bioreactor. Bioenergy Research 2011, 4:134-140.
• [23]Ramos LP, Breuil C, Saddler JN: The use of enzyme recycling and the influence of sugar accumulation on cellulose hydrolysis by Trichoderma-Cellulases. Enzyme Microb Technol 1993, 15:19-25.
• [24]Van Dyk JS, Pletschke BI: A review of lignocellulose bioconversion using enzymatic hydrolysis and synergistic cooperation between enzymes-Factors affecting enzymes, conversion and synergy. Biotechnol Adv 2012, 30:1458-1480.
• [25]Weiss N, Borjesson J, Pedersen LS, Meyer AS: Enzymatic lignocellulose hydrolysis: Improved cellulase productivity by insoluble solids recycling. Biotechnology for Biofuels 2013, 6:5. BioMed Central Full Text
• [26]Chirico WJ, Brown RD: β-Glucosidase from Trichoderma reesei. Eur J Biochem 1987, 165:343-351.
• [27]Tu MB, Chandra RP, Saddler JN: Recycling cellulases during the hydrolysis of steam exploded and ethanol pretreated lodgepole pine. Biotechnol Prog 2007, 23:1130-1137.
• [28]Varnai A, Viikari L, Marjamaa K, Siika-aho M: Adsorption of monocomponent enzymes in enzyme mixture analyzed quantitatively during hydrolysis of lignocellulose substrates. Bioresour Technol 2011, 102:1220-1227.
• [29]Qing Q, Yang B, Wyman CE: Xylooligomers are strong inhibitors of cellulose hydrolysis by enzymes. Bioresour Technol 2010, 101:9624-9630.
• [30]Pan XJ, Xie D, Gilkes N, Gregg DJ, Saddler JN: Strategies to enhance the enzymatic hydrolysis of pretreated softwood with high residual lignin content. Appl Biochem Biotechnol 2005, 121:1069-1079.
• [31]Taherzadeh MJ, Karimi K: Pretreatment of lignocellulosic wastes to improve ethanol and biogas production: A review. Int J Mol Sci 2008, 9:1621-1651.
• [32]Qi BK, Chen XR, Su Y, Wan YH: Enzyme adsorption and recycling during hydrolysis of wheat straw lignocellulose. Bioresour Technol 2011, 102:2881-2889.
• [33]Zhang JH, Tang M, Viikari L: Xylans inhibit enzymatic hydrolysis of lignocellulosic materials by cellulases. Bioresour Technol 2012, 121:8-12.
• [34]Sewalt VJH, Glasser WG, Beauchemin KA: Lignin Impact on Fiber Degradation. 3. Reversal of Inhibition of Enzymatic Hydrolysis by Chemical Modification of Lignin and by Additives. J Agric Food Chem 1997, 45:1823-1828.
• [35]Yang B, Wyman CE: BSA treatment to enhance enzymatic hydrolysis of cellulose in lignin containing substrates. Biotechnol Bioeng 2006, 94:611-617.
• [36]Zhang J, Lynd LR: Ethanol production from paper sludge by simultaneous saccharification and co-fermentation using recombinant xylose-fermenting microorganisms. Biotechnol Bioeng 2010, 107:235-244.
• [37]Yang J, Zhang X, Yong Q, Yu S: Three-stage enzymatic hydrolysis of steam-exploded corn stover at high substrate concentration. Bioresour Technol 2011, 102:4905-4908.
• [38]Qi BK, Luo JQ, Chen GQ, Chen XR, Wan YH: Application of ultrafiltration and nanofiltration for recycling cellulase and concentrating glucose from enzymatic hydrolyzate of steam exploded wheat straw. Bioresour Technol 2012, 104:466-472.
• [39]Humbird D, Davis R, Tao L, Kinchin C, Hsu D, Aden A, Schoen P, Lukas J, Olthof B, Worley M: Process design and economics for biochemical conversion of lignocellulosic biomass to ethanol. NREL Technical report. In NREL/TP-5100-47764. NREL, Golden: CO, USA; 2011. http://www.nrel.gov/biomass/pdfs/47764.pdf webcite
• [40]Sluiter A, Hames B, Ruiz R, Scarlata C, Sluiter J, Templeton D, Crocker D: Determination of Structural Carbohydrates and Lignin in Biomass. CO, USA: NREL, Golden; 2011. http://www.nrel.gov/biomass/analytical_procedures.html#lap-009 webcite
• [41]Kitcherside MA, Glen EF, Webster AJF: FibreCap: an improved method for the rapid analysis of fibre in feeding stuffs. Anim Feed Sci Technol 2000, 86:125-132.
• [42]Pribowo A, Arantes V, Saddler JN: The adsorption and enzyme activity profiles of specific Trichoderma reesei cellulase/xylanase components when hydrolyzing steam pretreated corn stover. Enzyme Microb Technol 2012, 50:195-203.
• [43]Ioelovich M, Morag E: Study of Enzymatic Hydrolysis of Pretreated Biomass at Increased Solids Loading. Bioresources 2012, 7:4672-4682.
• [44]Rodrigues AC, Leitão AF, Moreira S, Felby C, Gama M: Recycling of cellulases in lignocellulosic hydrolysates using alkaline elution. Bioresour Technol 2012, 110:526-533.
• [45]Wang QQ, Zhu JY, Hunt CG, Zhan HY: Kinetics of adsorption, desorption, and re-adsorption of a commercial endoglucanase in lignocellulosic suspensions. Biotechnol Bioeng 2012, 109:1965-1975.
• [46]Xue Y, Jameel H, Park S: Strategies to Recycle Enzymes and Their Impact on Enzymatic Hydrolysis for Bioethanol Production. Bioresources 2012, 7:602-615.
• [47]Tu MB, Chandra RP, Saddler JN: Evaluating the distribution of cellulases and the recycling of free cellulases during the hydrolysis of lignocellulosic substrates. Biotechnol Prog 2007, 23:398-406.
• [48]Bu LT, Nimlos MR, Shirts MR, Stahlberg J, Himmel ME, Crowley MF, Beckham GT: Product Binding Varies Dramatically between Processive and Nonprocessive Cellulase Enzymes. J Biol Chem 2012, 287:24807-24813.
• [49]Várnai A, Siika-aho M, Viikari L: Restriction of the enzymatic hydrolysis of steam-pretreated spruce by lignin and hemicellulose. Enzyme Microb Technol 2010, 46:185-193.
• [50]Hu JG, Arantes V, Saddler JN: The enhancement of enzymatic hydrolysis of lignocellulosic substrates by the addition of accessory enzymes such as xylanase: is it an additive or synergistic effect? Biotechnology for Biofuels 2011, 4:36. BioMed Central Full Text
• [51]Zhang X, Qin W, Paice MG, Saddler JN: High consistency enzymatic hydrolysis of hardwood substrates. Bioresour Technol 2009, 100:5890-5897.
• [52]Zhang Y, Liu YY, Xu JL, Yuan ZH, Qi W, Zhuang XS, He MC: High Solid and Low Enzyme Loading Based Saccharification of Agricultural Biomass. Bioresources 2012, 7:345-353.
• [53]Xue Y, Jameel H, Phillips R, Chang HM: Split addition cif enzymes in enzymatic hydrolysis at high solids concentration to increase sugar concentration for bioethanol production. J Ind Eng Chem 2012, 18:707-714.
• [54]Connor MR, Liao JC: Microbial production of advanced transportation fuels in non-natural hosts. Curr Opin Biotechnol 2009, 20:307-315.
• [55]Ghose TK: Measurement of Cellulase Activities. Pure Appl Chem 1987, 59:257-268.
• [56]Davis MW: A rapid modified method for compositional carbohydrate analysis of lignocellulosics by high pH anion-exchange chromatography with pulsed amperometric detection (HPAEC/PAD). J Wood Chem Technol 1998, 18:235-252.