Comparative assessment of acids and alkali based pretreatment on sawdust for enhanced saccharification with thermophilic cellulases

  • I. Haq
  • Z. Mustafa
  • A. Nawaz
  • H. Mukhtar
  • X. Zhou
  • Y. Xu
Keywords: Green energy, Renewable energy, Biofuel, Catalysis, Bioconversion

Abstract

The present study focuses on the use of thermophilic recombinant cellulases to produce fermentable sugars for conversion to bioethanol which is an important renewable energy resource to be worked upon due to climatic change, energy insecurity and nonrenewable nature of fossil fuels. Therefore, sawdust of pinus wood was subjected to pretreatment using different acids such as phosphoric, nitric, acetic and alkalis i.e. sodium hydroxide, calcium hydroxide and ammonia in various concentrations. Maximum delignification was observed using 8% nitric acid as it resulted in to 74.02% delignification and increased the cellulose availability to 81.8%. Subsequently, pretreated biomass was assessed for improvement in hydrolysis to less complex sugars employing thermophilic recombinant cellulases cloned from Thermotoga petrophila. Saccharification reaction parameters such as Incubation time, temperature, biomass and enzyme concentrations were optimized. The optimized conditions were revealed as 3 h incubation time, 65℃ temperature 0.1 % (w/v) substrate, 250, 2550 and 70140 U of Endo-1,4-β-glucanase, Exo-1,4-β-glucanase and β-1,4-Glucosidase, respectively. This optimization study resulted in 34.61% saccharification yield which is 1.82 folds increase compared to saccharification yield of untreated biomass. This study is unique in providing insight to pretreatment using Nitric acid as in literature use of nitric acid as a pretreatment agent is not well investigated.

References

Aguado, R., Lourenço, A.F., Ferreira, P.J., Moral, A. and Tijero, A. (2019). The relevance of the pretreatment on the chemical modification of cellulosic fibers. Cellulose. 2019 May 20:1-2. https://link.springer.com/article/10.1007/s10570-019-02517-7
Akhtar, N., Goyal, D. and Goyal, A. (2017). Characterization of microwave-alkali-acid pre-treated rice straw for optimization of ethanol production via simultaneous saccharification and fermentation (SSF). Energy Conversion and Management. Jun 1;141:133-44. https://www.sciencedirect.com/science/article/pii/S0196890416305702
Alrumman, S,A. (2016) Enzymatic saccharification and fermentation of cellulosic date palm wastes to glucose and lactic acid. Braz J Micrbiol. 47, no. 1: 110-119. https://www.sciencedirect.com/science/article/pii/S1517838215000167
Altavilla, G., Marinoni, S., Pancino, E., Galleti, S., Ragaini, S., Bellazzini, M., Cocozza, G., Bragaglia, A., Carrasco, J.M., Castro, A. and Di Fabrizio, L. (2015). The Gaia spectrophotometric standard stars survey: II. Instrumental effects of six ground‐based observing campaigns. Astronomische Nachrichten. Jul;336(6):515-29. https://onlinelibrary.wiley.com/doi/abs/10.1002/asna.201512176
Annamalai, N., Rajeswari, M.V. and Sivakumar, N. (2016). Cellobiohydrolases: role, mechanism, and recent developments. InMicrobial Enzymes in Bioconversions of Biomass (pp. 29-35). Springer, Cham. https://link.springer.com/chapter/10.1007/978-3-319-43679-1_2
Auxenfans, T.S., Buchoux, D., Larcher, G., Husson, E., Husson. and Sarazin, C. (2014) Enzymatic saccharification and structural properties of industrial wood sawdust: recycled ionic liquids pretreatments. Energy Convers. Manag. 88: 1094-1103. https://www.cabdirect.org/cabdirect/abstract/20153012959
Balat, M., Balat, H. and Öz, C. (2008) Progress in bioethanol processing. Prog. Energy Combust. Sci. 34,no.5:551-573. https://www.lth.se/fileadmin/bioteknik/Utbildning/Kurser/KKKA05/ProgressETOHprocess.pdf
Behera, S., Arora, R., Nandhagopal, N. and Kumar, S., 2014. Importance of chemical pretreatment for bioconversion of lignocellulosic biomass. Renewable and sustainable energy reviews, 36, pp.91-106. https://doi.org/10.1016/j.rser.2014.04.047
Bibi, R., Ahmad, Z., Imran, M., Hussain, S., Ditta, A., Mahmood, S. and Khalid, A. (2017). Algal bioethanol production technology: a trend towards sustainable development. Renewable and Sustainable Energy Reviews. May 1;71:976-85. https://www.sciencedirect.com/science/article/pii/S1364032116311819
Cara, C., Ruiz, E., Oliva, J.M., Sáez, F. and Castro, E., 2008. Conversion of olive tree biomass into fermentable sugars by dilute acid pretreatment and enzymatic saccharification. Bioresource technology, 99(6), pp.1869-1876. https://doi.org/10.1016/j.biortech.2007.03.037
Castoldi, R., Bracht, A., de Morais, G.R., Baesso, M.L., Correa, R.C., Peralta, R.A., Moreira, R.D., de Moraes, M.D., de Souza, C.G. and Peralta, R.M. (2014). Biological pretreatment of Eucalyptus grandis sawdust with white-rot fungi: study of degradation patterns and saccharification kinetics. Chemical Engineering Journal. Dec 15;258:240-6. https://www.sciencedirect.com/science/article/pii/S1385894714009851
Chen, H. and Jin, S. (2006). Effect of ethanol and yeast on cellulase activity and hydrolysis of crystalline cellulose. Enz Microb Technol. 39, no. 7: 1430-1432. https://www.researchgate.net/publication/238299730_Effect_of_ethanol_and_yeast_on_cellulase_activity_and_hydrolysis_of_crystalline_cellulose
Chen, H., Liu, J., Chang, X., Chen, D., Xue, Y., Liu, P., Lin, H. and Han, S. (2017). A review on the pretreatment of lignocellulose for high-value chemicals. Fuel Processing Technology.Jun 1;160:196-206. https://www.sciencedirect.com/science/article/pii/S0378382016311766
Colussi, F., Da Silva, V.M., Miller, I., Cota, J., de Oliveira, L.C., de Oliveira., Neto, M., Squina, F. M. and Garcia, W. (2015). Oligomeric state and structural stability of two hyperthermophilic β-glucosidases from Thermotoga petrophila. Amino acids. May 1;47(5):937-48. https://link.springer.com/article/10.1007/s00726-015-1923-3
Coughlan, M.P. and Mayer, F. (1992) The cellulase decomposing bacteria and their enzyme systems, In: Balowes A, Trurer H, Dworkin M, Harder W, Schleifer KH, (Eds.). The Prokaryotes, 2nd edn. Vol.-I. Springer Verlag: 460-516. https://www.researchgate.net/publication/226522753_Lignocellulose-Decomposing_Bacteria_and_Their_Enzyme_Systems
Duncan, E.G., O’Sullivan, C.A., Roper, M.M., Biggs, J.S. and Peoples, M.B. (2018). Influence of co-application of nitrogen with phosphorus, potassium and sulphur on the apparent efficiency of nitrogen fertiliser use, grain yield and protein content of wheat. Field Crops Research. Sep 1;226:56-65. https://www.sciencedirect.com/science/article/pii/S0378429018305987
Eklund, R., Galbe, M. and Zacchi. (1990). Optimization of temperature and enzyme concentration in the enzymatic saccharification of steam-pretreated willow, Enzyme. Microb. Tech. 12, no. 3: 225-228. https://www.sciencedirect.com/science/article/pii/014102299090043P
Guo, H., Chang, Y. and Lee, D.J., 2018. Enzymatic saccharification of lignocellulosic biorefinery: research focuses. Bioresource technology, 252, pp.198-215. https://doi.org/10.1016/j.biortech.2017.12.062
Haq, I., Arshad, Y., Nawaz, A., Aftab, M., Rehman, A., Mukhtar, H., Mansoor, Z. and Syed, Q. (2018). Removal of phenolic compounds through overliming for enhanced saccharification of wheat straw. J. Chem. Tech. Biotech. 93, no. 10: 3011-3017. https://onlinelibrary.wiley.com/doi/abs/10.1002/jctb.5659?af=R
Heck, R.H., Tabata, L. and Thomas, S.L. (2013). Multilevel and longitudinal modeling with IBM SPSS; Routledge. https://www.crcpress.com/Multilevel-and-Longitudinal-Modeling-with-IBM-SPSS/Heck-Thomas-Tabata/p/book/9780415817110
Hreggvidsson, G.O., Kaiste, E., Holst, O., Eggertsson, G., Palsdottir, A. and Kristjansson, J.K. (1996). An extremely thermostable cellulase from the thermophilic eubacterium Rhodothermus marinus. App.Environ.Microbial. 62,no.8:3047-3049. https://aem.asm.org/content/62/8/3047
Hsu, T.A. (2018). Pretreatment of biomass. InHandbook on bioethanol May 2 (pp. 179-212). Routledge. https://www.taylorfrancis.com/books/e/9780203752456/chapters/10.1201/9780203752456-10
Imran, M., Anwar, Z., Irshad, M., Asad, M.J. and Ashfaq, H. (2016). Cellulase production from species of fungi and bacteria from agricultural wastes and its utilization in industry: A review. Advances in Enzyme Research.;4(02):44. https://file.scirp.org/pdf/AER_2016052714523811.pdf
Khare, S.K., Pandey, A. and Larroche, C., 2015. Current perspectives in enzymatic saccharification of lignocellulosic biomass. Biochemical Engineering Journal, 102, pp.38-44. https://doi.org/10.1016/j.bej.2015.02.033
Kim, I., Lee, B., Park, J.Y., Choi, S.A. and Han, J.I. (2014). Effect of nitric acid on pretreatment and fermentation for enhancing ethanol production of rice straw. Carbohydrate polymers. 99: 563-567. https://www.ncbi.nlm.nih.gov/pubmed/24274544
Kim, J.K., Oh, B.R., Shin, H.J., Eom, C.Y. and Kim, S.W. (2008). Statistical optimization of enzymatic saccharification and ethanol fermentation using food waste, Process. Biochem. 43, no. 11: 1308-1312. https://www.sciencedirect.com/science/article/pii/S135951130800216X
Kim, N.J., Li, H., Jiang, M., Kang, J.W. and Chang, H.N. (2009). Simultaneous saccharification and fermentation of lignocellulosic residues pretreated with phosphoric acid–acetone for bioethanolproduction. Bioresour.Tech.100,no.13:3245-3251. https://www.ncbi.nlm.nih.gov/pubmed/19289273
Kim, S.B. and Lee, Y.Y. (1996). Fractionation of herbaceous biomass by ammonia-hydrogen peroxide percolation treatment. Appl. Biochem. Biotechnol. 57, no. 1: 147-156. https://link.springer.com/article/10.1007/BF02941695
Kim, T.H., Choi, C.H. and Oh, K.K. (2013). Bioconversion of sawdust into ethanol using dilute sulfuric acid-assisted continuous twin screw-driven reactor pretreatment and fed-batch simultaneous saccharification and fermentation. Bioresour. Technol. 130: 306-313. https://www.ncbi.nlm.nih.gov/pubmed/23306134
Kocher, G.S. and Kalra, K.L. (2013). Optimization of pretreatment, enzymatic saccharification and fermentation conditions for bioethanol production from rice straw. Ind. J. Appl. Res. 3, no. 5:62-64. https://www.researchgate.net/publication/285622182_Optimization_of_Pretreatment_Enzymatic_Saccharification_and_Fermentation_Conditions_for_Bioethanol_Production_from_Rice_Straw
Kumar, A.K. and Sharma, S. (2017). Recent updates on different methods of pretreatment of lignocellulosic feedstocks: a review. Bioresour. Bioprocess. 4. no.1:7. https://bioresourcesbioprocessing.springeropen.com/articles/10.1186/s40643-017-0137-9
Kumar, P., Barrett, D.M., Delwiche, M.J. and Stroeve, P. (2009). Methods for pretreatment of lignocellulosic biomass for efficient hydrolysis and biofuel production. Ind. Eng. Chem. Res. 48, no. 8: 3713-3729. https://pubs.acs.org/doi/10.1021/ie801542g
Kuusk, S. and Väljamäe, P. (2017). When substrate inhibits and inhibitor activates: implications of β-glucosidases. Biotechnology for biofuels. Dec;10(1):7. https://biotechnologyforbiofuels.biomedcentral.com/articles/10.1186/s13068-016-0690-z
Li, Y., Ruan, R., Chen, P.L., Liu, Z., Pan, X., Lin, X., Liu, Y., Mok, C.K. and Yang, T. (2004). Enzymatic hydrolysis of corn stover pretreated by combined dilute alkaline treatment and homogenization. Trans ASAE. 47, no. 3: 821. http://agris.fao.org/agris-search/search.do?recordID=US201300942034
Lim, W.S. and Lee, J.W. (2013). Influence of pretreatment condition on the fermentable sugar production and enzymatic hydrolysis of dilute acid-pretreated mixed softwood. Bioresour.Technol.140:306-311. https://www.ncbi.nlm.nih.gov/pubmed/23708848
Miller, G.L. (1959). Miller, Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem; 31(3): 426-428. https://pubs.acs.org/doi/10.1021/ac60147a030
Naidu, D.S., Hlangothi, S.P. and John, M.J. (2018). Bio-based products from xylan: A review. Carbohydrate polymers. 2018 Jan 1;179:28-41. https://www.sciencedirect.com/science/article/pii/S0144861717310974
Parisutham, V., Chandran, S.P., Mukhopadhyay, A., Lee, S.K. and Keasling, J.D. (2017). Intracellular cellobiose metabolism and its applications in lignocellulose-based biorefineries. Bioresource technology. Sep 1;239:496-506. https://www.sciencedirect.com/science/article/pii/S0960852417306399
Rastogi, M. and Shrivastava, S. (2017). Recent advances in second generation bioethanol production: An insight to pretreatment, saccharification and fermentation processes. Renewable and Sustainable Energy Reviews. Dec 1;80:330-40. https://www.sciencedirect.com/science/article/pii/S1364032117308651
Ravikumar, C., Kumar, P.S., Subhashni, S.K., Tejaswini, P.V. and Varshini, V. (2017). Microwave assisted fast pyrolysis of corn cob, corn stover, saw dust and rice straw: Experimental investigation on bio-oil yield and high heating values. Sustainable materials and technologies. Apr 1;11:19-27. https://www.sciencedirect.com/science/article/pii/S2214993716300951
Saqib, A.A., Hassan, M., Khan, N.F. and Baig, S. (2010). Thermostability of crude endoglucanase from Aspergillus fumigatus grown under solid state fermentation (SSF) and submerged fermentation (SmF). Process. Biochem. 45, no. 5:641-646. https://www.researchgate.net/publication/222238443_Thermostability_of_crude_endoglucanase_from_Aspergillus_fumigatus_grown_under_solid_state_fermentation_SSF_and_submerged_fermentation_SmF
Selig, M.J., Viamajala, S., Decker, S.R., Tucker, M.P., Himmel, M.E. and Vinzant, T.B. (2007). Deposition of lignin droplets produced during dilute acid pretreatment of maize stems retards enzymatic hydrolysis of cellulose. Biotechnol. Prog. 23, no. 6: 1333-1339. https://www.ncbi.nlm.nih.gov/pubmed/17973399
Taniguchi, M., Suzuki, H., Watanbe, D., Sakai, K., Hoshino, K. and Tanaka, T. (2005). Evaluation of pretreatment with Pleurotus ostreatus for enzymatic hydrolysis of rice straw. J. Biosci. Bioeng. 100, no. 6: 637-643. https://www.ncbi.nlm.nih.gov/pubmed/16473773
TAPPI Standard T236 cm-85 (1993) Kappa Number of Pulp. Tappi Journal, USA. https://research.cnr.ncsu.edu/wpsanalytical/documents/T236.PDF
Venu, H. and Madhavan, V. (2017). Effect of diethyl ether and Al2O3 nano additives in diesel-biodiesel-ethanol blends: Performance, combustion and emission characteristics. Journal of Mechanical Science and Technology. Jan 1;31(1):409-20. https://link.springer.com/article/10.1007/s12206-016-1243-x
Viikari, L., Alapuranen, M., Puranen, T., Vehmaanperä, J. and Siika-Aho, M. (2007). Thermostable enzymes in lignocellulose hydrolysis. In Biofuels, pp. 121-145. Springer, Berlin, Heidelberg. https://www.ncbi.nlm.nih.gov/pubmed/17589813
Viikari, L., Vehmaanperä, J. and Koivula, A. (2012). Lignocellulosic ethanol: from science to industry, Biomass.Bioenergy.46:13-24. https://www.researchgate.net/publication/257421146_Lignocellulosic_ethanol_From_science_to_industry
Xiang, Z., Chen, Y., Liu, Q. and Lu, F. (2018). A highly recyclable dip-catalyst produced from palladium nanoparticle-embedded bacterial cellulose and plant fibers. Green chemistry. 2018;20(5):1085-94. https://pubs.rsc.org/en/content/articlelanding/2018/gc/c7gc02835k/unauth#!divAbstract
Xiao, Z.Z., Zhang, X., Gregg, D. and Saddler, J. (2004). Effects of sugar inhibition on cellulases and b-glucosidase during enzymatic hydrolysis of softwood substrates, Appl. Biochem. Biotechnol. Breckenridge, CO, pp. 1115-1126. Humana Press, Totowa, NJ, 2004. https://link.springer.com/chapter/10.1007/978-1-59259-837-3_90
Xio, Z.Z., Zhang, X., Gregg, D. and Saddler, J. (2004). Effects of sugar inhibition on cellulases and b-glucosidase during enzymatic hydrolysis of softwood substrates, Appl. Biochem. Biotechnol, In Proceedings of the Twenty-Fifth Symposium on Biotechnology for Fuels and Chemicals Held May 4–7, in Breckenridge, CO, pp. 1115-1126. Humana Press, Totowa, NJ, 2004. https://link.springer.com/chapter/10.1007/978-1-59259-837-3_90
Yu, M., Li, J., Chang, S., Zhang, L., Mao, Y., Cui, T., Yan, Z., Luo, C. and Li, S. (2016). Bioethanol production using the sodium hydroxide pretreated sweet sorghum bagasse without washing. Fuel. Jul 1;175:20-5. https://www.sciencedirect.com/science/article/pii/S0016236116001332
Zhang, K.D., Li, W., Wang, Y.F., Zheng, Y.L., Tan, F.C., Ma, X.Q., Yao, L.S., Bayer, E.A, Wang, L.S. and Li, F.L. Processive degradation of crystalline cellulose by a multimodular endoglucanase via a wirewalking mode. Biomacromolecules. 2018 Apr 4;19(5):1686-96. https://pubs.acs.org/doi/abs/10.1021/acs.biomac.8b00340
Published
2020-07-23
How to Cite
Haq, I., Mustafa, Z., Nawaz, A., Mukhtar, H., Zhou, X., & Xu, Y. (2020). Comparative assessment of acids and alkali based pretreatment on sawdust for enhanced saccharification with thermophilic cellulases. Revista Mexicana De Ingeniería Química, 19(Sup. 1), 305-314. https://doi.org/10.24275/rmiq/Bio1702
Section
Biotechnology

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