Cellobiohydrolase and endoglucanase respond differently to surfactants during the hydrolysis of cellulose

Biotechnology for Biofuels

Chia-wen C Hsieh²  David Cannella²  Henning Jørgensen¹  Claus Felby²  Lisbeth G Thygesen² 

[1] Present address: Center for Bioprocess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Søltofts Plads, Building 229, Kgs. Lyngby, DK-2800, Denmark

[2] Department of Geosciences and Natural Resource Management, Faculty of Science, University of Copenhagen, Rolighedsvej 23, Frederiksberg C, DK-1958, Denmark

关键词: Water constraint;    PASC hydrolysis;    Avicel hydrolysis;    Monocomponent cellulase hydrolysis;    Enzymatic saccharification of cellulose;    Surfactants;    PEG;   

Others  :  1145260
DOI  :  10.1186/s13068-015-0242-y

 received in 2014-09-10, accepted in 2015-03-19,  发布年份 2015

【 摘 要 】

Background

Non-ionic surfactants such as polyethylene glycol (PEG) can increase the glucose yield obtained from enzymatic saccharification of lignocellulosic substrates. Various explanations behind this effect include the ability of PEG to increase the stability of the cellulases, decrease non-productive cellulase adsorption to the substrate, and increase the desorption of enzymes from the substrate. Here, using lignin-free model substrates, we propose that PEG also alters the solvent properties, for example, water, leading the cellulases to increase hydrolysis yields.

Results

The effect of PEG differs for the individual cellulases. During hydrolysis of Avicel and PASC with a processive monocomponent exo-cellulase cellobiohydrolase (CBH) I, the presence of PEG leads to an increase in the final glucose concentration, while PEG caused no change in glucose production with a non-processive endoglucanase (EG). Also, no effect of PEG was seen on the activity of β-glucosidases. While PEG has a small effect on the thermostability of both cellulases, only the activity of CBH I increases with PEG. Using commercial enzyme mixtures, the hydrolysis yields increased with the addition of PEG. In parallel, we observed that the relaxation time of the hydrolysis liquid phase, as measured by LF-NMR, directly correlated with the final glucose yield. PEG was able to boost the glucose production even in highly concentrated solutions of up to 150 g/L of glucose.

Conclusions

The hydrolysis boosting effect of PEG appears to be specific for CBH I. The mechanism could be due to an increase in the apparent activity of the enzyme on the substrate surface. The addition of PEG increases the relaxation time of the liquid-phase water, which from the data presented points towards a mechanism related to PEG-water interactions rather than PEG-protein or PEG-substrate interactions.

【 授权许可】

   
2015 Hsieh et al.; licensee BioMed Central.

【 图 表 】

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【 参考文献 】

• [1]Ooshima H, Sakata M, Harano Y: Enhancement of enzymatic hydrolysis of cellulose by surfactant. Biotechnol Bioeng 1986, 28:1727-34.
• [2]Duff SJB, Murray WD: Bioconversion of forest products industry waste cellulosics to fuel ethanol: a review. Bioresour Technol 1996, 55:1-33.
• [3]Sun Y, Cheng J: Hydrolysis of lignocellulosic materials for ethanol production: a review. Bioresour Technol 2002, 83:1-11.
• [4]Börjesson J, Peterson R, Tjerneld F: Enhanced enzymatic conversion of softwood lignocellulose by poly(ethylene glycol) addition. Enzyme Microb Technol 2007, 40:754-62.
• [5]Kristensen JB, Börjesson J, Bruun MH, Tjerneld F, Jørgensen H: Use of surface active additives in enzymatic hydrolysis of wheat straw lignocellulose. Enzyme Microb Technol 2007, 40:888-95.
• [6]Kaar WE, Holtzapple MT: Benefits from Tween during enzymic hydrolysis of corn stover. Biotechnol Bioeng 1998, 59:419-27.
• [7]Tu M, Zhang X, Paice M, McFarlane P, Saddler JN: Effect of surfactants on separate hydrolysis fermentation and simultaneous saccharification fermentation of pretreated lodgepole pine. Biotechnol Prog 2009, 25:1122-9.
• [8]Cannella D, Jørgensen H: Do new cellulolytic enzyme preparations affect the industrial strategies for high solids lignocellulosic ethanol production? Biotechnol Bioeng 2014, 111:59-68.
• [9]Börjesson J, Engqvist M, Sipos B, Tjerneld F: Effect of poly(ethylene glycol) on enzymatic hydrolysis and adsorption of cellulase enzymes to pretreated lignocellulose. Enzyme Microb Technol 2007, 41:186-95.
• [10]Eriksson T, Börjesson J, Tjerneld F: Mechanism of surfactant effect in enzymatic hydrolysis of lignocellulose. Enzyme Microb Technol 2002, 31:353-64.
• [11]Chylenski P, Felby C, Haven MØ, Gama M, Selig MJ: Precipitation of Trichoderma reesei commercial cellulase preparations under standard enzymatic hydrolysis conditions for lignocelluloses. Biotechnol Lett 2012, 34:1475-82.
• [12]Reese ET: Inactivation of cellulase by shaking and its prevention by surfactants. J Appl Biochem 1980, 2:36-9.
• [13]Park JW, Takahata Y, Kajiuchi T, Akehata T: Effects of nonionic surfactant on enzymatic hydrolysis of used newspaper. Biotechnol Bioeng 1992, 39:117-20.
• [14]Li J, Li S, Fan C, Yan Z: The mechanism of poly(ethylene glycol) 4000 effect on enzymatic hydrolysis of lignocellulose. Colloids Surf B Biointerfaces 2012, 89:203-10.
• [15]Helle SS, Duff SJB, Cooper DG: Effect of surfactants on cellulose hydrolysis. Biotechnol Bioeng 1993, 42:611-7.
• [16]Zhang M, Ouyang J, Liu B, Yu H, Jiang T, Cai C, et al.: Comparison of hydrolysis efficiency and enzyme adsorption of three different cellulosic materials in the presence of poly(ethylene glycol). BioEnergy Res 2013, 6:1252-9.
• [17]Ouyang J, Dong Z, Song X, Lee X, Chen M, Yong Q: Improved enzymatic hydrolysis of microcrystalline cellulose (Avicel PH101) by polyethylene glycol addition. Bioresour Technol 2010, 101:6685-91.
• [18]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, 28:1478-90.
• [19]Hsieh CC, Cannella D, Jørgensen H, Felby C, Thygesen LG: Cellulase inhibition by high concentrations of monosaccharides. J Agric Food Chem 2014, 62:3800-5.
• [20]Chundawat SPS, Lipton MS, Purvine SO, Uppugundla N, Gao D, Balan V, et al.: Proteomics-based Compositional analysis of complex Cellulase–Hemicellulase mixtures. J Proteome Res 2012, 10:4365-72.
• [21]Horn SJ, Sikorski P, Cederkvist JB, Vaaje-Kolstad G, Sørlie M, Synstad B, et al.: Costs and benefits of processivity in enzymatic degradation of recalcitrant polysaccharides. Proc Natl Acad Sci 2006, 103:18089-94.
• [22]Kurašin M, Väljamäe P: Processivity of cellobiohydrolases is limited by the substrate. J Biol Chem 2011, 286:169-77.
• [23]Igarashi K, Uchihashi T, Koivula A, Wada M, Kimura S, Okamoto T, et al.: Traffic jams reduce hydrolytic efficiency of cellulase on cellulose surface. Science 2011, 333:1279-82.
• [24]Várnai 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, 102:1220-7.
• [25]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.
• [26]Bu L, Nimlos MR, Shirts MR, Ståhlberg J, Himmel ME, Crowley MF, et al.: Product binding varies dramatically between processive and nonprocessive cellulase enzymes. J Biol Chem 2012, 287:24807-13.
• [27]Skovgaard PA, Jørgensen H: Influence of high temperature and ethanol on thermostable lignocellulolytic enzymes. J Ind Microbiol Biotechnol 2013, 40:447-56.
• [28]Reese ET, Mandels M: Stability of the cellulase of Trichoderma reesei under use conditions. Biotechnol Bioeng 1980, 22:323-35.
• [29]Merino ST, Cherry J: Progress and challenges in enzyme development for biomass utilization. In Biofuels. Springer Berlin, Heidelberg; 2007:95-120.
• [30]Allen AL, Mortensen RE: Production of cellulase from trichoderma reesei in fed-batch fermentation from soluble carbon sources. Biotechnol Bioeng 1981, 23:2641-5.
• [31]Ghose TK: Measurement of cellulase activities. Pure Appl Chem 1987, 59:257-68.
• [32]Saha BC, Bothast RJ: Production, purification, and characterization of a highly glucose-tolerant novel beta-glucosidase from Candida peltata. Appl Environ Microbiol 1996, 62:3165-70.
• [33]Starcher B: A ninhydrin-based assay to quantitate the total protein content of tissue samples. Anal Biochem 2001, 292:125-9.
• [34]Walseth CS: Occurrence of cellulases in enzyme preparations from microorganisms. Tappi 1952, 35:228-33.
• [35]Westereng B, Agger JW, Horn SJ, Vaaje-Kolstad G, Aachmann FL, Stenstrøm YH, et al.: Efficient separation of oxidized cello-oligosaccharides generated by cellulose degrading lytic polysaccharide monooxygenases. J Chromatogr A 2013, 1271:144-52.
• [36]Provencher SW: CONTIN: a general purpose constrained regularization program for inverting noisy linear algebraic and integral equations. Comput Phys Commun 1982, 27:229-42.