Enzymatic Hydrolysis of Cellulose with Different Crystallinities Studied by Means of SEC-MALLS

›› 2011, Vol. 19 ›› Issue (5): 773-778.

• SELECTED PAPERS FROM THE 4TH NATIONAL CONFERENCE ON MASS TRANSFER AND SEPARATION ENGINEERING AND IN HONOUR OF PROF. K. T. YU (YU GUOCONG) • Previous Articles Next Articles

Enzymatic Hydrolysis of Cellulose with Different Crystallinities Studied by Means of SEC-MALLS

ZHANG Mingjia, SU Rongxin, QI Wei, DU Ruoyu, HE Zhimin

State Key Laboratory of Chemical Engineering, Chemical Engineering Research Center, School of Chemical Engi-neering and Technology, Tianjin University, Tianjin 300072, China

Received:2011-06-10 Revised:2011-07-13 Online:2011-10-28 Published:2011-10-28
Supported by:
Supported by the National Natural Science Foundation of China (20976130 and 20806057);National Science and Technology Pillar Program of China (2007BAD42B02);Program for New Century Excellent Talents in University of Ministry of Education of China (No.NCET-08-0386);the R&D program of Tianjin Binhai New Area (2010-BK17C004)

Enzymatic Hydrolysis of Cellulose with Different Crystallinities Studied by Means of SEC-MALLS

张名佳, 苏荣欣, 齐崴, 杜若愚, 何志敏

State Key Laboratory of Chemical Engineering, Chemical Engineering Research Center, School of Chemical Engi-neering and Technology, Tianjin University, Tianjin 300072, China

通讯作者: HE Zhimin,E-mail:enzyme@tju.edu.cn
基金资助:
Supported by the National Natural Science Foundation of China (20976130 and 20806057);National Science and Technology Pillar Program of China (2007BAD42B02);Program for New Century Excellent Talents in University of Ministry of Education of China (No.NCET-08-0386);the R&D program of Tianjin Binhai New Area (2010-BK17C004)

Abstract

Abstract: The reactions of exo-cellulase (cellobiohydrolase, CBH) and endo-cellulase (endoglucanase, EG) were investigated by analyzing the insoluble residues of microcrystalline cellulose (MCC) and filter paper cellulose (FPC) during enzymatic hydrolysis. Molecular parameters including molecular weight and its distribution, degree of po-lymerization, and radii of gyration were measured by size exclusion chromatography coupled with multi-angle laser light scattering. No significant change in MCC chains was found during the whole reaction period, indicating that CBH digestion follows a layer-by-layer solubilization manner. This reaction mode might be the major reason for slow enzymatic hydrolysis of cellulose. On the other hand, the degree of polymerization of FPC chains decreases rapidly in the initial reaction, indicating that EG digestion follows a random scission manner, which may create new ends for CBH easily. The slopes of the conformation plots for MCC and FPC increase gradually, indicating stronger chain stiffness of cellulose during hydrolysis.

Key words: cellulose, cellobiohydrolase, endoglucanase, relative molecular mass distribution, SEC-MALLS-DRI

摘要： The reactions of exo-cellulase (cellobiohydrolase, CBH) and endo-cellulase (endoglucanase, EG) were investigated by analyzing the insoluble residues of microcrystalline cellulose (MCC) and filter paper cellulose (FPC) during enzymatic hydrolysis. Molecular parameters including molecular weight and its distribution, degree of po-lymerization, and radii of gyration were measured by size exclusion chromatography coupled with multi-angle laser light scattering. No significant change in MCC chains was found during the whole reaction period, indicating that CBH digestion follows a layer-by-layer solubilization manner. This reaction mode might be the major reason for slow enzymatic hydrolysis of cellulose. On the other hand, the degree of polymerization of FPC chains decreases rapidly in the initial reaction, indicating that EG digestion follows a random scission manner, which may create new ends for CBH easily. The slopes of the conformation plots for MCC and FPC increase gradually, indicating stronger chain stiffness of cellulose during hydrolysis.

关键词: cellulose, cellobiohydrolase, endoglucanase, relative molecular mass distribution, SEC-MALLS-DRI

ZHANG Mingjia, SU Rongxin, QI Wei, DU Ruoyu, HE Zhimin. Enzymatic Hydrolysis of Cellulose with Different Crystallinities Studied by Means of SEC-MALLS[J]. , 2011, 19(5): 773-778.

张名佳, 苏荣欣, 齐崴, 杜若愚, 何志敏. Enzymatic Hydrolysis of Cellulose with Different Crystallinities Studied by Means of SEC-MALLS[J]. , 2011, 19(5): 773-778.

References

1 Béguin, P., Aubert, J.P., “The biological degradation of cellulose”, FEMS Microbiology Reviews, 13, 25-58 (1994).
2 Lima, M.M.D., Borsali, R., “Rodlike cellulose microcrystals:Structure, properties, and applications”, Macromol. Rapid Commun., 25, 771-787 (2004).
3 Sjöholm, E., Gustafsson, K., Eriksson, B., Brown, W., Colmsjo, A., “Aggregation of cellulose in lithium chloride/N,N-dimethylacetamide”, Carbohydr. Polym., 41, 153-161 (2000).
4 Zhang, M.J., Su, R.X., Qi, W., He, Z.M., “Enhanced enzymatic hydrolysis of lignocellulose by optimizing enzyme complexes”, Appl. Biochem. Biotechnol., 160, 1407-1414 (2010).
5 Gusakov, A.V., Salanovich, T.N., Antonov, A.I., Ustinov, B.B., Okunev, O.N., Burlingame, R., Emalfarb, M., Baez, M., Sinitsyn, A.P., “Design of highly efficient cellulase mixtures for enzymatic hydrolysis of cellulose”, Biotechnol. Bioeng., 97, 1028-1038 (2007).
6 Eremeeva, T., “Size-exclusion chromatography of enzymatically treated cellulose and related polysaccharides:a review”, J. Biochem. Bioph. Methods, 56, 253-264 (2003).
7 Eremeeva, T., Bikova, T., Eisimonte, M., Viesturs, U., Treimanis, A., “Fractionation and molecular characteristics of cellulose during enzymatic hydrolysis”, Cellulose, 8, 69-79 (2001).
8 Chen, Y., Stipanovic, A.J., Winter, W.T., Wilson, D.B., Kim, Y.J., “Effect of digestion by pure cellulases on crystallinity and average chain length for bacterial and microcrystalline celluloses”, Cellulose, 14, 283-293 (2007).
9 Pala, H., Mota, M., Gama, F.M., “Enzymatic depolymerisation of cellulose”, Carbohydr. Polym., 68, 101-108 (2007).
10 KlemanLeyer, K.M., SiikaAho, M., Teeri, T.T., Kirk, T.K., “The cellulases endoglucanase I and cellobiohydrolase Ⅱ of Trichoderma reesei act synergistically to solubilize native cotton cellulose but not to decrease its molecular size”, Applied and Environmental Microbiology, 62, 2883-2887 (1996).
11 Srisodsuk, M., Kleman-Leyer, K., Keranen, S., Kirk, T.K., Teeri, T.T., “Modes of action on cotton and bacterial cellulose of a homologous endoglucanase-exoglucanase pair from Trichoderma reesei”, Eur. J. Biochem., 251, 885-892 (1998).
12 Gupta, R., Lee, Y.Y., “Mechanism of cellulase reaction on pure cellulosic substrates”, Biotechnol. Bioeng., 102, 1570-81 (2009).
13 Klemanleyer, K.M., Gilkes, N.R., Miller, R.C., Kirk, T.K., “Changes in the molecular-size distribution of insoluble celluloses by the action of recombinant cellulomonas-fimi cellulases”, Biochem. J, 302, 463-469 (1994).
14 Zhang, Y.H.P., Himmel, M.E., Mielenz, J.R., “Outlook for cellulase improvement:Screening and selection strategies”, Biotechnol. Adv., 24, 452-481 (2006).
15 Josefsson, P., Henriksson, G., Wagberg, L., “The physical action of cellulases revealed by a quartz crystal microbalance study using ultrathin cellulose films and pure”, Biomacromolecules, 9, 249-254 (2008).
16 Ottøy, M.H., Vårum, K.M., Christensen, B.E., Anthonsen, M.W., Smidsr d, O., “Preparative and analytical size-exclusion chromatography of chitosans”, Carbohydr. Polym., 31, 253-261 (1996z).
17 Wyatt, P.J., “Light scattering and the absolute characterization of macromolecules”, Anal. Chim. Acta, 272, 1-40 (1993).
18 Andersson, M., Wittgren, B., Wahlund, K.G., “Accuracy in multiangle light scattering measurements for molar mass and radius estimations. Model calculations and experiments”, Anal. Chem., 75, 4279-4291 (2003).
19 Schult, T., Hjerde, T., Inge Optun, O., Kleppe, P.J., Moe, S., “Characterization of cellulose by SEC-MALLS”, Cellulose, 9, 149-158 (2002).
20 Yanagisawa, M., Shibata, I., Isogai, A., “SEC-MALLS analysis of cellulose using LiCl/1,3-dimethyl-2-imidazolidinone as an eluent”, Cellulose, 11, 169-176 (2004).
21 Berggren, R., Berthold, F., Sjoholm, E., Lindstrom, M., “Improved methods for evaluating the molar mass distributions of cellulose in Kraft pulp”, J. Appl. Polym. Sci., 88, 1170-1179 (2003).
22 Yanagisawa, M., Isogai, A., “SEC-MALS-QELS study on the molecular conformation of cellulose in LiCl/amide solutions”, Biomacromolecules, 6, 1258-1265 (2005).
23 Wood, T.M., Bhat, K.M., “Methods for measuring cellulase activities”, Methods Enzymol., 160, 87-112 (1988).
24 Zhang, Y.H.P., Lynd, L.R., “Toward an aggregated understanding of enzymatic hydrolysis of cellulose:Noncomplexed cellulase systems”, Biotechnol. Bioeng., 88, 797-824 (2004).
25 Andersen, N., Johansen, K.S., Michelsen, M., Stenby, E.H., Krogh, K., Olsson, L., “Hydrolysis of cellulose using mono-component enzymes shows synergy during hydrolysis of phosphoric acid swollen cellulose (PASC), but competition on Avicel”, Enzyme Microb. Technol., 42, 362-370 (2008).
26 Mansfield, S.D., Meder, R., “Cellulose hydrolysis-the role of monocomponent cellulases in crystalline cellulose degradation”, Cellulose, 10, 159-169 (2003).
27 van Wyk, J.P.H., “Paper hydrolysis by cellulase from penicillium funiculosum and Trichoderma viride”, Bioresour. Technol., 63, 275-277 (1998).
28 Eriksson, T., Karlsson, J., Tjerneld, F., “A model explaining declining rate in hydrolysis of lignocellulose substrates with cellobiohydrolase I (Cel7A) and endoglucanase I (Cel7B) of Trichoderma reesei”, Appl. Biochem. Biotechnol., 101, 41-60 (2002).
29 Goh, K.K.T., Pinder, D.N., Hall, C.E., Hemar, Y., “Rheological and light scattering properties of flaxseed polysaccharide aqueous solutions”, Biomacromolecules, 7, 3098-3103 (2006).

[1]	Jie Wei, Weiwei Yang, Shuai Jia, Jie Wei, Ziqiang Shao. N, P co-doped porous graphene with high electrochemical properties obtained via the laser induction of cellulose nanofibrils [J]. Chinese Journal of Chemical Engineering, 2022, 47(7): 31-38.
[2]	Shaoxiang Cai, Han Yan, Qiuyi Wang, He Han, Ru Li, Zhichao Lou. Top-down strategy for bamboo lignocellulose-derived carbon heterostructure with enhanced electromagnetic wave dissipation [J]. Chinese Journal of Chemical Engineering, 2022, 43(3): 360-369.
[3]	Zhongxin Lin, Renliang Huang, Jiangjiexing Wu, Anastasia Penkova, Wei Qi, Zhimin He, Rongxin Su. Injectable self-healing nanocellulose hydrogels crosslinked by aluminum: Cellulose nanocrystals vs. cellulose nanofibrils [J]. Chinese Journal of Chemical Engineering, 2022, 50(10): 389-397.
[4]	Xiujuan Li, Le Chen, Dandan Zhu, Song Yang, Zhong Wu, Mingyang He, Zhihui Zhang, Qun Chen. Preparation of hybridizing zeolitic imidazolate frameworks with carboxymethylcellulose for adsorption separation of n-hexane/3-methylpentane [J]. Chinese Journal of Chemical Engineering, 2021, 29(1): 103-109.
[5]	Lan Lan, Yan Shao, Yilai Jiao, Rongxin Zhang, Christopher Hardacre, Xiaolei Fan. Systematic study of H₂ production from catalytic photoreforming of cellulose over Pt catalysts supported on TiO₂ [J]. Chinese Journal of Chemical Engineering, 2020, 28(8): 2084-2091.
[6]	Hairong Yuan, Ruolin Guan, Akiber Chufo Wachemo, Yatian Zhang, Xiaoyu Zuo, Xiujin Li. Improving physicochemical characteristics and anaerobic digestion performance of rice straw via ammonia pretreatment at varying concentrations and moisture levels [J]. Chinese Journal of Chemical Engineering, 2020, 28(2): 541-547.
[7]	Xingyilong Zhang, Houfang Lu, Kejing Wu, Yingying Liu, Changjun Liu, Yingming Zhu, Bin Liang. Hydrolysis of mechanically pre-treated cellulose catalyzed by solid acid SO₄^2--TiO₂ in water-ethanol solvent [J]. Chinese Journal of Chemical Engineering, 2020, 28(1): 136-142.
[8]	Meisam Sharifi, Seyed-Mortaza Robatjazi, Minoo Sadri, Jafar Mohammadian Mosaabadi. Immobilization of organophosphorus hydrolase enzyme by covalent attachment on modified cellulose microfibers using different chemical activation strategies: Characterization and stability studies [J]. Chin.J.Chem.Eng., 2019, 27(1): 191-199.
[9]	Md. Sakinul Islam, Nhol Kao, Sati N. Bhattacharya, Rahul Gupta, Hyoung Jin Choi. Potential aspect of rice husk biomass in Australia for nanocrystalline cellulose production [J]. Chin.J.Chem.Eng., 2018, 26(3): 465-476.
[10]	Chih-Heng Wang, Wen-Hua Chen, Hwai-Shen Liu, Jinn-Tsyy Lai, Cheng-Che Hsu, Ben-Zu Wan. Process development for producing a food-grade glucose solution from rice straws [J]. Chin.J.Chem.Eng., 2018, 26(2): 386-392.
[11]	Dongsheng Xue, Xuhao Zeng, Chunjie Gong, Dongqiang Lin, Shanjing Yao. A cold adapt and ethanol tolerant endoglucanase from a marine Bacillus subtilis [J]. Chin.J.Chem.Eng., 2018, 26(12): 2601-2606.
[12]	Kibrom Alebel Gebru, Chandan Das. Effects of solubility parameter differences among PEG,PVP and CA on the preparation of ultrafiltration membranes:Impacts of solvents and additives on morphology,permeability and fouling performances [J]. , 2017, 25(7): 911-923.
[13]	Changdong Li, Carlos Amador, Yulong Ding. Effect of carboxymethyl cellulose on dissolution kinetics of carboxymethyl cellulose-sodium carbonate two-component tablet [J]. , 2017, 25(10): 1545-1550.
[14]	Changdong Li, Carlos Amador, Yulong Ding. Effect of carboxymethyl cellulose on dissolution kinetics of carboxymethyl cellulose-sodium carbonate two-component tablet [J]. , 2017, 25(10): 1545-1550.
[15]	Yan Li, Hongxian Fan, Xueqing Yu, Songmei Zhang, Gang Li. Hemicellulose in corn straw: Extracted from alkali solution and produced 5-hydroxymethyl furfural in HCOOH/HCOONa buffer solution [J]. , 2016, 24(12): 1786-1792.

Enzymatic Hydrolysis of Cellulose with Different Crystallinities Studied by Means of SEC-MALLS

Enzymatic Hydrolysis of Cellulose with Different Crystallinities Studied by Means of SEC-MALLS

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments