A new strategy for biological enzyme bleaching: combined effects of laccase, xylanase, and mannanase in the bleaching of softwood kraft pulp – a synergistic effect of enzymes
Source abstract
Abstract
Abstract Although highly efficient, conventional chemical bleaching is criticized for its high cost and environmental impact. In contrast, enzyme bleaching offers a sustainable and eco-friendly alternative, especially in elemental chlorine-free (ECF) and total chlorine-free (TCF) processes. This study investigates the synergy of laccase, xylanase, and mannanase in the ECF bleaching of softwood kraft pulp, focusing on their effects on fiber morphology and bleaching performance. The results demonstrate that laccase effectively oxidizes lignin, enhancing pulp brightness, while xylanase and mannanase improve bleaching efficiency by breaking down hemicellulose. The addition of mannanase during the enzymatic treatment increased the pulp brightness by 1.58 %ISO, with almost no negative impact on physical properties. The synergistic effect of the three enzymes increased the pulp brightness by 5.37 %ISO, reaching a maximum brightness of 92.35 %ISO. Furthermore, mannanase treatment significantly reduces the consumption of chemicals, cutting laccase and violuric acid usage by 38 % or decreasing total chlorine dioxide use by 13.26 %, while maintaining the same brightness level. These findings highlight the potential of using laccase, xylanase, and mannanase for efficient and eco-friendly pulp bleaching, contributing to resource conservation and environmental benefits. This study provides valuable insights into the industrial application of enzyme-based bleaching for sustainable paper production.
Source: crossref · Quality: baseline
Research context
OpenAlex topics: Enzyme-mediated dye degradation · Lignin and Wood Chemistry · Biochemical and biochemical processes
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
State Key Laboratory of Biobased Fiber Manufacturing Technology , 66345 Tianjin University of Science and Technology , Tianjin , 300457 , China
Tianjin Key Laboratory of Pulp and Paper , Tianjin University of Science and Technology , Tianjin , 300457 , China
Funding
the National Key Research and Development Program of China
References (44)
Show source-provided references
1. Afsahi, G., Rojalin, T., and Vuorinen, T. (2019) Chemical characteristics and stability of eucalyptus kraft pulps bleached with tertiary amine catalyzed hypochlorous acid. Cellulose 26:2047-2054, https://doi.org/10.1007/s10570-018-2172-x.10.1007/s10570-018-2172-x ↗
2. Almeida, N., Meyer, V., Burnet, A., Boucher, J., Talens-Perales, D., Pereira, S., Ihalainen, P., Levée, T., Polaina, J., Petit-Conil, M., et al.. (2022). Use of a novel extremophilic xylanase for an environmentally friendly industrial bleaching of kraft pulps. Int. J. Mol. Sci. 23: 13423, https://doi.org/10.3390/ijms232113423.10.3390/ijms232113423 ↗
3. Anand, G., Leibman-Markus, M., Elkabetz, D., and Bar, M. (2021). Method for the production and purification of plant immuno-active xylanase from trichoderma. Int. J. Mol. Sci. 22: 4214, https://doi.org/10.3390/ijms22084214.10.3390/ijms22084214 ↗
4. Angural, S., Rana, M., Sharma, A., Warmoota, R., Puri, N., and Gupta, N. (2020). Combinatorial biobleaching of mixedwood pulp with lignolytic and hemicellulolytic enzymes for paper making. Indian J. Microbiol. 60: 383–387, https://doi.org/10.1007/s12088-020-00867-6.10.1007/s12088-020-00867-6 ↗
5. Angural, S., Bala, I., Kumar, A., Kumar, D., Jassal, S., and Gupta, N. (2021). Bleach enhancement of mixed wood pulp by mixture of thermo-alkali-stable xylanase and mannanase derived through co-culturing of Alkalophilic bacillus sp. NG-27 and Bacillus nealsonii PN-11. Heliyon 7: e05673, https://doi.org/10.1016/j.heliyon.2020.e05673.10.1016/j.heliyon.2020.e05673 ↗
6. Benetti, F., Lemos, C.A.A., de Oliveira Gallinari, M., Terayama, A.M., Briso, A.L.F., de Castilho Jacinto, R., Sivieri-Araújo, G., and Cintra, L.T.A. (2018). Influence of different types of light on the response of the pulp tissue in dental bleaching: a systematic review. Clin. Oral Invest. 22: 1825–1837, https://doi.org/10.1007/s00784-017-2278-9.10.1007/s00784-017-2278-9 ↗
7. Briganti, L., Manzine, L.R., de Mello Capetti, C.C., de Araújo, E.A., de Oliveira Arnoldi Pellegrini, V., Guimaraes, F.E.G., de Oliveira Neto, M., and Polikarpov, I. (2024). Unravelling biochemical and structural features of Bacillus licheniformis GH5 mannanase using site-directed mutagenesis and high-resolution protein crystallography studies. Int. J. Biol. Macromol. 274: 133182, https://doi.org/10.1016/j.ijbiomac.2024.133182.10.1016/j.ijbiomac.2024.133182 ↗
8. Dhawan, S. and Kaur, J. (2007). Microbial mannanases: an overview of production and applications. Crit. Rev. Biotechnol. 27: 197–216, https://doi.org/10.1080/07388550701775919.10.1080/07388550701775919 ↗
9. Gonçalves, I., Herrero-Yniesta, V., Perales Arce, I., Escrigas Castañeda, M., Cavaco-Paulo, A., and Silva, C. (2014). Ultrasonic pilot-scale reactor for enzymatic bleaching of cotton fabrics. Ultrason. Sonochem. 21: 1535–1543, https://doi.org/10.1016/j.ultsonch.2014.02.009.10.1016/j.ultsonch.2014.02.009 ↗
10. Handoko, O.T., Hasanudin, U., Suroso, E., Dermiyati, D., Yuwono, S.D., Ginting, S.B., Sugiharto, R., Indraningtyas, L., Amelia, J.R., and Iryani, D.A. (2024). Economic and environmental analysis of spent bleaching earth reactivation. IOP Conf. Ser. Earth Environ. Sci. 1308: 012061, https://doi.org/10.1088/1755-1315/1308/1/012061.10.1088/1755-1315/1308/1/012061 ↗
11. Hutterer, C., Kliba, G., Punz, M., Fackler, K., and Potthast, A. (2017). Enzymatic pulp upgrade for producing high-value cellulose out of a kraft paper pulp. Enzyme Microb. Technol. 102: 67–73, https://doi.org/10.1016/j.enzmictec.2017.03.014.10.1016/j.enzmictec.2017.03.014 ↗
12. Janusz, G., Pawlik, A., Świderska-Burek, U., Polak, J., Sulej, J., Jarosz-Wilkołazka, A., and Paszczyński, A. (2020). Laccase properties, physiological functions, and evolution. Int. J. Mol. Sci. 21: 966, https://doi.org/10.3390/ijms21030966.10.3390/ijms21030966 ↗
13. Kanwal, N., Rashid, N., Irfan Majeed, M., Nawaz, H., Amber, A., Zohaib, M., Bano, A., Albekairi, N.A., Alshammari, A., Shahzadi, A., et al.. (2025). Surface-enhanced Raman spectroscopy for the characterization of xylanases enzyme. Spectrochim. Acta Mol. Biomol. Spectrosc. 325: 125065, https://doi.org/10.1016/j.saa.2024.125065.10.1016/j.saa.2024.125065 ↗
14. Kapoor, M., Kapoor, R.K., and Kuhad, R.C. (2007). Differential and synergistic effects of xylanase and laccase mediator system (LMS) in bleaching of soda and waste pulps. J. Appl. Microbiol. 103: 305–317, https://doi.org/10.1111/j.1365-2672.2006.03251.x.10.1111/j.1365-2672.2006.03251.x ↗
15. Kaur, D., Bhardwaj, N.K., and Lohchab, R.K. (2018). A study on pulping of rice straw and impact of incorporation of chlorine dioxide during bleaching on pulp properties and effluents characteristics. J. Clean. Prod. 170: 174–182, https://doi.org/10.1016/j.jclepro.2017.09.111.10.1016/j.jclepro.2017.09.111 ↗
16. Kaur, R., Salwan, R., and Sharma, V. (2022). Structural properties, genomic distribution of laccases from streptomyces and their potential applications. Process Biochem. 118: 133–144, https://doi.org/10.1016/j.procbio.2022.04.015.10.1016/j.procbio.2022.04.015 ↗
17. Kim, J.K., Lim, S.H., and Kang, H.W. (2013). Cloning and characterization of a novel laccase gene, fvlac7, based on the genomic sequence of flammulina velutipes. Mycobiology 41: 37–41, https://doi.org/10.5941/myco.2013.41.1.37.10.5941/MYCO.2013.41.1.37 ↗
18. Ko, C.H., Chen, F.J., Lee, J.J., and Tzou, D.L.M. (2011). Effects of fiber physical and chemical characteristics on the interaction between endoglucanase and eucalypt fibers. Cellulose 18: 1043–1054, https://doi.org/10.1007/s10570-011-9534-y.10.1007/s10570-011-9534-y ↗
19. Liu, J., Deng, J., Tan, X., Li, Y., Li, H., Cheng, W., and Jiang, Z. (2025). Co-integration of laccase and xylanase from bacillus pumilus into mini-cellulosome facilitates softwood sulfite pulp biobleaching and reduces hydrogen peroxide consumption. Int. J. Biol. Macromol. 290: 139048, https://doi.org/10.1016/j.ijbiomac.2024.139048.10.1016/j.ijbiomac.2024.139048 ↗
20. Loi, M., Glazunova, O., Fedorova, T., Logrieco, A.F., and Mulè, G. (2021). Fungal laccases: the forefront of enzymes for sustainability. J. Fungi 7: 1048, https://doi.org/10.3390/jof7121048.10.3390/jof7121048 ↗
21. Martín-Sampedro, R., Rodríguez, A., Ferrer, A., García-Fuentevilla, L., and Eugenio, M. (2012). Biobleaching of pulp from oil palm empty fruit bunches with laccase and xylanase. Bioresour. Technol. 110: 371–378, https://doi.org/10.1016/j.biortech.2012.01.111.10.1016/j.biortech.2012.01.111 ↗
22. Meesupthong, R., Yingkamhaeng, N., Nimchua, T., Pinmanee, P., Mussatto, S.I., Li, B., and Sukyai, P. (2021). Xylanase pretreatment of energy cane enables facile cellulose nanocrystal isolation. Cellulose 28: 799–812, https://doi.org/10.1007/s10570-020-03559-y.10.1007/s10570-020-03559-y ↗
23. Monje, P.G., González-García, S., Moldes, D., Vidal, T., Romero, J., Moreira, M., and Feijoo, G. (2010). Biodegradability of kraft mill TCF biobleaching effluents: application of enzymatic laccase-mediator system. Water Res. 44: 2211–2220, https://doi.org/10.1016/j.watres.2009.12.047.10.1016/j.watres.2009.12.047 ↗
24. Nagpal, R., Bhardwaj, N.K., and Mahajan, R. (2020). Synergistic approach using ultrafiltered xylano-pectinolytic enzymes for reducing bleaching chemical dose in manufacturing rice straw paper. Environ. Sci. Pollut. Res. 27: 44637–44646, https://doi.org/10.1007/s11356-020-11104-4.10.1007/s11356-020-11104-4 ↗
25. Nie, S., Liu, X., Wu, Z., Zhan, L., Yin, G., Yao, S., Song, H., and Wang, S. (2014). Kinetics study of oxidation of the lignin model compounds by chlorine dioxide. Chem. Eng. J. 241: 410–417, https://doi.org/10.1016/j.cej.2013.10.068.10.1016/j.cej.2013.10.068 ↗
26. Parab, P. and Khandeparker, R. (2021). Xylanolytic enzyme consortia from bacillus sp. NIORKP76 for improved biobleaching of kraft pulp. Bioprocess Biosyst. Eng. 44: 2513–2524, https://doi.org/10.1007/s00449-021-02623-6.10.1007/s00449-021-02623-6 ↗
27. Patel, K., Vaghamshi, N., Shah, K., Duggirala, S.M., Ghelani, A., Dudhagara, P., and Shyu, D.J.H. (2024). Synergistic use of thermostable laccase and xylanase in optimizing the pre-bleaching of kraft pulp. Catalysts 14: 1, https://doi.org/10.3390/catal14010001.10.3390/catal14010001 ↗
28. Quintana, E., Valls, C., Vidal, T., and Blanca Roncero, M. (2013). An enzyme-catalyzed bleaching treatment to meet dissolving pulp characteristics for cellulose derivatives applications. Bioresour. Technol. 148: 1–8, https://doi.org/10.1016/j.biortech.2013.08.104.10.1016/j.biortech.2013.08.104 ↗
29. Quintana, E., Valls, C., Barneto, A.G., Vidal, T., Ariza, J., and Roncero, M.B. (2015). Studying the effects of laccase treatment in a softwood dissolving pulp: cellulose reactivity and crystallinity. Carbohydr. Polym. 119: 53–61, https://doi.org/10.1016/j.carbpol.2014.11.019.10.1016/j.carbpol.2014.11.019 ↗
30. Ravalason, H., Bertaud, F., Herpoël-Gimbert, I., Meyer, V., Ruel, K., Joseleau, J.P., Grisel, S., Olivé, C., Sigoillot, J.C., and Petit-Conil, M. (2012). Laccase/HBT and laccase-CBM/HBT treatment of softwood kraft pulp: impact on pulp bleachability and physical properties. Bioresour. Technol. 121: 68–75, https://doi.org/10.1016/j.biortech.2012.06.077.10.1016/j.biortech.2012.06.077 ↗
31. Sharma, N., Bhardwaj, N.K., and Singh, R.B.P. (2020). Environmental issues of pulp bleaching and prospects of peracetic acid pulp bleaching: a review. J. Clean. Prod. 256: 120338, https://doi.org/10.1016/j.jclepro.2020.120338.10.1016/j.jclepro.2020.120338 ↗
32. Sharma, D., Nagpal, R., Agrawal, S., Bhardwaj, N., and Mahajan, R. (2022). Eco-friendly bleaching of agrowaste wheat straw using crude alkalo-thermotolerant cellulase-free xylano-pectinolytic enzymes. Appl. Biochem. Biotechnol. 194: 620–634, https://doi.org/10.1007/s12010-021-03641-6.10.1007/s12010-021-03641-6 ↗
33. Singh, R., Kumar, M., Mittal, A., and Mehta, P.K. (2016). Microbial enzymes: industrial progress in 21st century. 3 Biotech 6: 174, https://doi.org/10.1007/s13205-016-0485-8.10.1007/s13205-016-0485-8 ↗
34. Song, H.T., Gao, Y., Yang, Y.M., Xiao, W.J., Liu, S.H., Xia, W.C., Liu, Z.L., Yi, L., and Jiang, Z.B. (2016). Synergistic effect of cellulase and xylanase during hydrolysis of natural lignocellulosic substrates. Bioresour. Technol. 219: 710–715, https://doi.org/10.1016/j.biortech.2016.08.035.10.1016/j.biortech.2016.08.035 ↗
35. Surma-Ślusarska, B. and Leks-Stępień, J. (2001). TCF bleaching of kraft pulps with laccase and xylanaseB. J. Wood Chem. Technol. 21: 361–370.10.1081/WCT-100108331 ↗
36. Uğur, Ş.S., Karaboyacı, M., and Boguniewicz-Zablocka, J. (2023). Influence of cotton bleaching methods on the parameters of generated textile industrial wastewater. Sustainability 15: 15592, https://doi.org/10.3390/su152115592.10.3390/su152115592 ↗
37. Valls, C. and Roncero, M.B. (2009). Using both xylanase and laccase enzymes for pulp bleaching. Bioresour. Technol. 100: 2032–2039, https://doi.org/10.1016/j.biortech.2008.10.009.10.1016/j.biortech.2008.10.009 ↗
38. Valls, C., Cadena, E.M., and Blanca Roncero, M. (2013). Obtaining biobleached eucalyptus cellulose fibres by using various enzyme combinations. Carbohydr. Polym. 92: 276–282, https://doi.org/10.1016/j.carbpol.2012.08.083.10.1016/j.carbpol.2012.08.083 ↗
39. Valls, C., Cusola, O., Vidal, T., Torres, A.L., and Roncero, M.B. (2019). A straightforward bioprocess for a cleaner paper decolorization. J. Clean. Prod. 236: 117702, https://doi.org/10.1016/j.jclepro.2019.117702.10.1016/j.jclepro.2019.117702 ↗
40. Walia, A., Guleria, S., Mehta, P., Chauhan, A., and Parkash, J. (2017). Microbial xylanases and their industrial application in pulp and paper biobleaching: a review. 3 Biotech 7: 11, https://doi.org/10.1007/s13205-016-0584-6.10.1007/s13205-016-0584-6 ↗
41. Witayakran, S. and Ragauskas, A.J. (2009). Modification of high-lignin softwood kraft pulp with laccase and amino acids. Enzyme Microb. Technol. 44: 176–181, https://doi.org/10.1016/j.enzmictec.2008.10.011.10.1016/j.enzmictec.2008.10.011 ↗
42. Ye, Z.-H. and Zhong, R. (2022). Outstanding questions on xylan biosynthesis. Plant Sci. 325: 111476, https://doi.org/10.1016/j.plantsci.2022.111476.10.1016/j.plantsci.2022.111476 ↗
43. Zhang, D., Li, X., Wang, M., Ye, Y., Du, J., Lu, X., and Zhao, J. (2016). Xylanase treatment suppresses light- and heat-induced yellowing of pulp. Sci. Rep. 6: 38374, https://doi.org/10.1038/srep38374.10.1038/srep38374 ↗
44. Zhi Fu, G., Chan, A., and Minns, D. (2005). Preliminary assessment of the environmental benefits of enzyme bleaching for pulp and paper making (7 pp). Int. J. Life Cycle Assess. 10: 136–142, https://doi.org/10.1065/lca2004.06.162.10.1065/lca2004.06.162 ↗