Advertisement

Cellulose

pp 1–17 | Cite as

The effect of hemicellulose on the binding and activity of cellobiohydrolase I, Cel7A, from Trichoderma reesei to cellulose

  • S. Malgas
  • V. M. Kwanya Minghe
  • B. I. PletschkeEmail author
Original Research
  • 39 Downloads

Abstract

Hydrothermal pre-treatments decrease lignocellulose recalcitrance against enzymatic hydrolysis by removing the majority of the hemicellulose, thus increasing cellulase accessibility. However, a small amount of the hemicellulose may remain and become adsorbed to the cellulose, leading to cellulase inhibition. Here, we produced hemicellulose bound cellulose, using glucuronoxylan and galactomannan, to simulate hydrothermally pre-treated hardwoods and softwoods, respectively, and evaluated how this can affect cellulose hydrolysis by Trichoderma reesei derived cellobiohydrolase I (Cel7A). Based on X-ray powder diffraction (XRD), histochemistry, scanning electron microscopy and Simon’s staining, hemicellulose binding onto cellulose affected the physical properties of the biomass, which subsequently affected its hydrolysis rate. As a result of hemicellulose binding onto cellulose, the adsorption of Cel7A was significantly impacted (up to 45%), leading to lowered activities (a 40% reduction), especially for glucuronoxylan. The bound hemicellulose may be released from the cellulose during agitation and hydrolysis. We therefore evaluated the effect of free hemicellulose on Cel7A. Free xylan was more inhibitory to Cel7A than free mannan, demonstrating non-competitive inhibition, while mannan exhibited uncompetitive inhibition. The recalcitrant effect of both bound and free hemicellulose could be relieved by the addition of hemicellulolytic enzymes (i.e. XT6 and Man26A) during cellulose hydrolysis. During the degradation of cellulose in “realistic” woody biomasses by Cel7A, the addition of hemicellulases led to a significant improvement in cellulose hydrolysis. This study showed that hemicellulose remains a critical factor regarding biomass recalcitrance and that the addition of hemicellulolytic activities in commercial enzyme cocktails is required (especially the mannanolytic activities lacking from most commercial enzyme cocktails), in order to realise high sugar yields at low enzyme protein loadings for low-cost biofuel production.

Graphic abstract

Keywords

Cel7A Cellulose hydrolysis Enzyme adsorption Hemicellulose Inhibition 

Notes

Acknowledgments

Financial support from the National Research Foundation (NRF) of South Africa (NRF Grant No. 96004) and Rhodes University (Sandisa Imbewu) is gratefully acknowledged. Any opinion, findings and conclusions or recommendations expressed in this material are those of the author(s) and therefore the NRF and CSIR do not accept any liability in regard thereto.

References

  1. Bååth JA, Abad AM, Berglund J et al (2018) Mannanase hydrolysis of Spruce galactoglucomannan focusing on the influence of acetylation on enzymatic mannan degradation. Biotechnol Biofuels 11:1–15CrossRefGoogle Scholar
  2. Beckham GT, Matthews JF, Bomble YJ et al (2010) Identification of amino acids responsible for processivity in a family 1 carbohydrate-binding module from a fungal cellulase. J Phys Chem B 114:1447–1453PubMedCrossRefPubMedCentralGoogle Scholar
  3. Bernardes A, Pellegrini VOA, Curtolo F et al (2019) Carbohydrate binding modules enhance cellulose enzymatic hydrolysis by increasing access of cellulases to the substrate. Carbohydr Polym 211:57–68PubMedCrossRefPubMedCentralGoogle Scholar
  4. Boraston AB, Bolam DN, Gilbert HJ, Davies GJ (2004) Carbohydrate-binding modules: fine-tuning polysaccharide recognition. Biochem J 382:769–781PubMedPubMedCentralCrossRefGoogle Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254Google Scholar
  6. Chandra RP, Saddler JN (2012) Use of the simons’ staining technique to assess cellulose accessibility in pretreated substrates. Ind Biotechnol 8:230–237CrossRefGoogle Scholar
  7. Chandra R, Ewanick S, Hsieh C, Saddler JN (2008) The characterization of pretreated lignocellulosic substrates prior to enzymatic hydrolysis. Biotechnol Prog 24:1178–1185PubMedCrossRefPubMedCentralGoogle Scholar
  8. Chandra RP, Ewanick SM, Chung PA et al (2009) Comparison of methods to assess the enzyme accessibility and hydrolysis of pretreated lignocellulosic substrates. Biotechnol Lett 31:1217–1222PubMedCrossRefPubMedCentralGoogle Scholar
  9. Esteghlalian AR, Bilodeau M, Mansfield SD, Saddler JN (2001) Do enzymatic hydrolyzability and Simons’ stain reflect the changes in the accessibility of lignocellulosic substrates to cellulase enzymes? Biotechnol Prog 17:1049–1054PubMedCrossRefPubMedCentralGoogle Scholar
  10. Filonova L, Kallas ÅM, Greffe L et al (2007) Analysis of the surfaces of wood tissues and pulp fibers using carbohydrate-binding modules specific for crystalline cellulose and mannan. Biomacromol 8:91–97CrossRefGoogle Scholar
  11. Goldstein MA, Takagi M, Hashida S et al (1993) Characterization of the cellulose-binding domain of the Clostridium cellulovorans cellulose-binding protein A. Society 175:5762–5768Google Scholar
  12. Gourlay K, Hu J, Arantes V et al (2015) The use of carbohydrate binding modules (CBMs) to monitor changes in fragmentation and cellulose fiber surface morphology during cellulase- and swollenin-induced deconstruction of lignocellulosic substrates. J Biol Chem 290:2938–2945PubMedCrossRefPubMedCentralGoogle Scholar
  13. Guo J, Catchmark JM (2013) Binding specificity and thermodynamics of cellulose-binding modules from Trichoderma reesei Cel7A and Cel6A. Biomacromol 14:1268–1277CrossRefGoogle Scholar
  14. Hébert-Ouellet Y, Meddeb-mouelhi F, Khatri V, Cui L et al (2017) Tracking and predicting wood fibers processing with fluorescent carbohydrate binding modules. Green Chem 19:2603–2611CrossRefGoogle Scholar
  15. Hilden L, Daniel G, Johansson G (2003) Use of a fluorescence labelled, carbohydrate-binding module from Phanerochaete chrysosporium Cel7D for studying wood cell wall ultrastructure. Biotechnol Lett 25:553–558PubMedCrossRefPubMedCentralGoogle Scholar
  16. Hu J, Saddler JN (2018) Why does GH10 xylanase have better performance than GH11 xylanase for the deconstruction of pretreated biomass? Biomass Bioenerg 110:13–16CrossRefGoogle Scholar
  17. Hu J, Arantes V, Saddler JN (2011) The enhancement of enzymatic hydrolysis of lignocellulosic substrates by the addition of accessory enzymes such as xylanase: is it an additive or synergistic effect? Biotechnol Biofuels 4:36PubMedPubMedCentralCrossRefGoogle Scholar
  18. Hu J, Chandra R, Arantes V et al (2015) The addition of accessory enzymes enhances the hydrolytic performance of cellulase enzymes at high solid loadings. Bioresour Technol 186:149–153PubMedCrossRefPubMedCentralGoogle Scholar
  19. Jalak J, Valjamae P (2014) Multi-mode binding of cellobiohydrolase Cel7A from Trichoderma reesei to cellulose. PLoS ONE 9:e108181PubMedPubMedCentralCrossRefGoogle Scholar
  20. Kazmi MZH, Karmakar A, Michaelis VK, Williams FJ (2019) Separation of cellulose/hemicellulose from lignin in white pine sawdust using boron trihalide reagents. Tetrahedron 75:1465–1470CrossRefGoogle Scholar
  21. Khatri V, Meddeb-mouelhi F, Beauregard M (2018) New insights into the enzymatic hydrolysis of lignocellulosic polymers by using fluorescent tagged carbohydrate-binding modules. Sustain Energy Fuels 2:479–491CrossRefGoogle Scholar
  22. Kohnke T, Brelid H, Westman G (2009) Adsorption of cationized barley husk xylan on kraft pulp fibres: influence of degree of cationization on adsorption characteristics. Cellulose 16:1109–1121CrossRefGoogle Scholar
  23. Kohnke T, Lund K, Brelid H, Westman G (2010) Kraft pulp hornification: a closer look at the preventive effect gained by glucuronoxylan adsorption. Carbohydr Polym 81:226–233CrossRefGoogle Scholar
  24. Le Costaouec T, Pakarinen A, Varnai A et al (2013) The role of carbohydrate binding module (CBM) at high substrate consistency: comparison of Trichoderma reesei and Themoascus aurantiacus Cel7A (CBHI) and Cel5A (EGII). Bioresour Technol 143:196–203PubMedCrossRefPubMedCentralGoogle Scholar
  25. Linder Å, Bergman R, Bodin A, Gatenholm P (2003) Mechanism of assembly of xylan onto cellulose surfaces. Langmuir 19:5072–5077CrossRefGoogle Scholar
  26. Long L, Tian D, Zhai R et al (2018) Thermostable xylanase-aided two-stage hydrolysis approach enhances sugar release of pretreated lignocellulosic biomass. Bioresour Technol 257:334–338PubMedCrossRefPubMedCentralGoogle Scholar
  27. Maeda H, Ishida N (1967) Specificity of binding of hexopyranosyl polysaccharides with Fluorescent Brightener. J Biochem 62:276–278PubMedCrossRefPubMedCentralGoogle Scholar
  28. Malgas S, van Dyk JS, Pletschke BI (2015) A review of the enzymatic hydrolysis of mannans and synergistic interactions between β-mannanase, β-mannosidase and α-galactosidase. World J Microbiol Biotechnol 31:1167–1175PubMedCrossRefPubMedCentralGoogle Scholar
  29. Malgas S, Susan Van Dyk J, Abboo S, Pletschke BI (2016) The inhibitory effects of various substrate pre-treatment by-products and wash liquors on mannanolytic enzymes. J Mol Catal B Enzym 123:132–140CrossRefGoogle Scholar
  30. Malgas S, Chandra R, Van Dyk JS et al (2017) Formulation of an optimized synergistic enzyme cocktail, HoloMix, for effective degradation of various pre-treated hardwoods. Bioresour Technol 245:52–65PubMedCrossRefPubMedCentralGoogle Scholar
  31. Martínez-abad A, Berglund J, Toriz G et al (2017) Regular motifs in xylan modulate molecular flexibility and interactions with cellulose surfaces. Plant Physiol 175:1579–1592PubMedPubMedCentralCrossRefGoogle Scholar
  32. Mitra PP, Loqué D (2014) Histochemical staining of Arabidopsis thaliana secondary cell wall elements. J Vis Exp 87:1–11Google Scholar
  33. Momeni MH, Ubhayasekera W, Sandgren M et al (2015) Structural insights into the inhibition of cellobiohydrolase Cel7A by xylo-oligosaccharides. FEBS J 282:2167–2177PubMedCrossRefPubMedCentralGoogle Scholar
  34. Park S, Baker JO, Himmel ME et al (2010) Cellulose crystallinity index: measurement techniques and their impact on interpreting cellulase performance. Biotechnol Biofuels 3:10PubMedPubMedCentralCrossRefGoogle Scholar
  35. Pellegrini VOA, Lei N, Kyasaram M et al (2014) Reversibility of substrate adsorption for the cellulases Cel7A, Cel6A, and Cel7B from Hypocrea jecorina. Langmuir 30:12602–12609PubMedCrossRefPubMedCentralGoogle Scholar
  36. Rahikainen JL, Evans JD, Mikander S et al (2013) Cellulase–lignin interactions-the role of carbohydrate-binding module and pH in non-productive binding. Enzyme Microb Technol 53:315–321PubMedCrossRefGoogle Scholar
  37. Rastogi M, Shrivastava S (2017) Recent advances in second generation bioethanol production: an insight to pretreatment, saccharification and fermentation processes. Renew Sustain Energy Rev 80:330–340CrossRefGoogle Scholar
  38. Rodríguez-Zúñiga UF, Cannella D, de Giordano RC et al (2015) Lignocellulose pretreatment technologies affect the level of enzymatic cellulose oxidation by LPMO. Green Chem 17:2896–2903CrossRefGoogle Scholar
  39. Saini JK, Saini R, Tewari L (2014) Lignocellulosic agriculture wastes as biomass feedstocks for second-generation bioethanol production: concepts and recent developments. 3 Biotech 5:337–353PubMedPubMedCentralCrossRefGoogle Scholar
  40. Segal L, Creely J, Martin A, Conrad C (1959) An empirical method for estimating the degree of crystallinity of native cellulose using the x-ray diffractometer. Text Res J 29:786–794CrossRefGoogle Scholar
  41. Selig MJ, Knoshaug EP, Adney WS et al (2008) Synergistic enhancement of cellobiohydrolase performance on pretreated corn stover by addition of xylanase and esterase activities. Bioresour Technol 99:4997–5005PubMedCrossRefPubMedCentralGoogle Scholar
  42. Selig MJ, Adney WS, Himmel ME, Decker SR (2009) The impact of cell wall acetylation on corn stover hydrolysis by cellulolytic and xylanolytic enzymes. Cellulose 16:711–722CrossRefGoogle Scholar
  43. Sluiter JB, Ruiz RO, Scarlata CJ et al (2010) Compositional analysis of lignocellulosic feedstocks. 1. Review and description of methods. J Agric Food Chem 58:9043–9053PubMedPubMedCentralCrossRefGoogle Scholar
  44. Srisodsuk M, Lehtig J, Linder M et al (1997) Trichoderma reesei cellobiohydrolase I with an endoglucanase cellulose-binding domain: action on bacterial microcrystalline cellulose. J Biotechnol 57:49–57PubMedCrossRefPubMedCentralGoogle Scholar
  45. Sun D, Alam A, Tu Y et al (2017) Steam-exploded biomass saccharification is predominantly affected by lignocellulose porosity and largely enhanced by Tween-80 in Miscanthus. Bioresour Technol 239:1–8CrossRefGoogle Scholar
  46. Thygesen A, Oddershede J, Lilholt H et al (2005) On the determination of crystallinity and cellulose content in plant fibres. Cellulose 12:563–576CrossRefGoogle Scholar
  47. Várnai A, Huikko L, Pere J et al (2011) Synergistic action of xylanase and mannanase improves the total hydrolysis of softwood. Bioresour Technol 102:9096–9104PubMedCrossRefPubMedCentralGoogle Scholar
  48. Von Freiesleben P, Spodsberg N, Stenbæk A et al (2018) Boosting of enzymatic softwood saccharification by fungal GH5 and GH26 endomannanases. Biotechnol Biofuels 11:1–14CrossRefGoogle Scholar
  49. Wang X, Li K, Yang M et al (2016a) Hydrolyzability of mannan after adsorption on cellulose. Cellulose 24:35–47CrossRefGoogle Scholar
  50. Wang X, Li K, Yang M, Zhang J (2016b) Hydrolyzability of xylan after adsorption on cellulose: exploration of xylan limitation on enzymatic hydrolysis of cellulose. Carbohydr Polym 148:362–370PubMedCrossRefPubMedCentralGoogle Scholar
  51. Whitney SEC, Brigham JE, Darke AH et al (1998) Structural aspects of the interaction of mannan-based polysaccharides with bacterial cellulose. Carbohydr Res 307:299–309CrossRefGoogle Scholar
  52. Wood PJ (1980) Specificity in the interaction of direct dyes with polysaccharides. Carbohydr Polym 85:271–287CrossRefGoogle Scholar
  53. Xin D, Yang M, Chen X, Zhang J (2017) Recovering activities of inactivated cellulases by the use of mannanase in spruce hydrolysis. ACS Sustain Chem Eng 5:5265–5272CrossRefGoogle Scholar
  54. Yeh T, Chang M, Chang W (2014) Comparison of dilute acid and sulfite pretreatments on Acacia confusa for biofuel application and the influence of its extractives. J Agric Food Chem 62:10768–10775PubMedCrossRefPubMedCentralGoogle Scholar
  55. Yu L, Lyczakowski JJ, Pereira C et al (2018) The patterned structure of galactoglucomannan suggests it may bind to cellulose in seed mucilage. Plant Physiol 178:1011–1026PubMedPubMedCentralCrossRefGoogle Scholar
  56. Zhai R, Hu J, Saddler JN (2016) What are the major components in steam pretreated lignocellulosic biomass that inhibit the efficacy of cellulase enzyme mixtures? ACS Sustain Chem Eng 4:3429–3436CrossRefGoogle Scholar
  57. Zhai R, Hu J, Saddler JN (2018) The inhibition of hemicellulosic sugars on cellulose hydrolysis are highly dependant on the cellulase productive binding, processivity, and substrate surface charges. Bioresour Technol 258:79–87PubMedCrossRefPubMedCentralGoogle Scholar
  58. Zhang Z, Donaldson AA, Ma X (2012) Advancements and future directions in enzyme technology for biomass conversion. Biotechnol Adv 30:913–919PubMedCrossRefPubMedCentralGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Enzyme Science Programme (ESP), Department of Biochemistry and MicrobiologyRhodes UniversityGrahamstownSouth Africa
  2. 2.Department of Chemical EngineeringUniversity of LorraineLorraineFrance

Personalised recommendations