Skip to main content
SpringerLink
Log in

Sequential pretreatment of sugarcane bagasse by alkali and organosolv for improved delignification and cellulose saccharification by chimera and cellobiohydrolase for bioethanol production

  • Original Article
  • Published:
3 Biotech Aims and scope Submit manuscript

Abstract

Sequential pretreatments for sugarcane bagasse (scb) by NaOH followed by organosolv under mild conditions were evaluated for cellulose recovery and dilignification. The best-optimized sequential pretreatment of scb was obtained at 10% (w/v) of raw scb loading at 1% (w/v) NaOH (50 °C, 2 h) followed by treatment with organosolv (85%, v/v phosphoric acid, 50 °C, 1 h) with chilled acetone. This sequentially pretreated scb showed cellulose recovery, 66.1% (w/w) and delignification, 83.2% (w/w). NaOH or organosolv pretreated scb showed lower cellulose recovery 47.4% (w/w) or 54.5% (w/w) with lower delignification, 61% (w/w) or 56% (w/w), respectively. Pretreated solid residue of sequentially pretreated scb was enzymatically saccharified by chimera (β-glucosidase and endoglucanase, CtGH1-L1-CtGH5-F194A) and cellobiohydrolase (CtCBH5A) cloned from Clostridium thermocellum. Enzymatic hydrolysate of best sequentially pretreated scb gave total reducing sugar (TRS) yield, 230 mg/g and glucose yield, 137 mg/g pretreated scb. Only organosolv pretreated scb gave TRS yield, 112.5 mg/g and glucose yield, 72 mg/g of pretreated scb. Thus, sequentially pretreated scb resulted in 37% higher enzymatic digestibility than only orgnaosolv pretreated scb. Higher enzymatic digestibility was supported by higher crystallinity index CrI (45%) than those obtained with only organosolv pretreated (38%) or raw scb (25%). Field Emission Scanning Electron Microscope (FESEM) and Fourier-transform infrared (FT-IR) analyses showed enhanced cellulose exposure in sequentially pretreated scb. Preliminary investigation of bioethanol production at small scale by separate hydrolysis and fermentation (SHF) of enzymatic hydrolysate from best sequentially pretreated scb by Saccharomyces cerevisiae gave maximum ethanol yield of 0.42 g/g of glucose.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Asadi N, Zilouei H (2017) Optimization of organosolv pretreatment of rice straw for enhanced biohydrogen production using Enterobacter aerogenes. Bioresour Technol 227:335–344

    Article  CAS  PubMed  Google Scholar 

  • Ascencio JJ, Chandel AK, Philippini RR, da Silva SS (2019) Comparative study of cellulosic sugars production from sugarcane bagasse after dilute nitric acid, dilute sodium hydroxide and sequential nitric acid-sodium hydroxide pretreatment. Biomass Convers Biorefin 10:813–822

    Article  Google Scholar 

  • Asgher M, Ahmad Z, Iqbal HMN (2013) Alkali and enzymatic delignification of sugarcane bagasse to expose cellulose polymers for saccharification and bio-ethanol production. Ind Crop Prod 44:488–495

    Article  CAS  Google Scholar 

  • Bak JS, Ko JK, Han YH, Lee BC, Choi IG, Kim KH (2009) Improved enzymatic hydrolysis yield of rice straw using electron beam irradiation pretreatment. Bioresour Technol 100:1285–1290

    Article  CAS  PubMed  Google Scholar 

  • Bekiaris G, Lindedam J, Peltre C, Decker SR, Turner GB, Magid J, Bruun S (2015) Rapid estimation of sugar release from winter wheat straw during bioethanol production using FTIR-photoacoustic spectroscopy. Biotechnol Biofuels 8:85

    Article  PubMed  PubMed Central  Google Scholar 

  • Bharadwaja STP, Singh S, Moholkar VS (2015) Design and optimization of a sono-hybrid process for bioethanol production from Parthenium hysterophorus. J Taiwan Inst Chem Eng 51:71–78

    Article  Google Scholar 

  • Bi S, Peng L, Chen K, Zhu Z (2016) Enhanced enzymatic saccharification of sugarcane bagasse pretreated by combining O2 and NaOH. Bioresour Technol 214:692–699

    Article  CAS  PubMed  Google Scholar 

  • Binod P, Satyanagalakshmi K, Sindhu R, Janu KU, Sukumaran RK, Pandey A (2012) Short duration microwave assisted pretreatment enhances the enzymatic saccharification and fermentable sugar yield from sugarcane bagasse. Renew Energy 37:109–116

    Article  CAS  Google Scholar 

  • Borah AJ, Singh S, Goyal A, Moholkar VS (2016) An assessment of the potential of invasive weeds as multiple feedstocks for biofuel production. RSC Adv 6:47151–47163

    Article  CAS  Google Scholar 

  • 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 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Cao S, Aita GM (2013) Enzymatic hydrolysis and ethanol yields of combined surfactant and dilute ammonia treated sugarcane bagasse. Bioresour Technol 131:357–364

    Article  CAS  PubMed  Google Scholar 

  • Chaikumpollert O, Methacanon P, Suchiva K (2004) Structural elucidation of hemicelluloses from Vetiver grass. Carbohydr Polym 57:191–196

    Article  CAS  Google Scholar 

  • Gupta A, Rajulapati V, Das D, Goyal A (2017) Comparative analysis of bioethanol production involving saccharification by mixed recombinant clostridial enzymes using sugarcane leaves and kans grass as sustainable feed stocks from North-east India. Indian J Biotechnol 16:199–210

    CAS  Google Scholar 

  • Gupta R, Aswal VK, Saini JK (2020) Sequential dilute acid and alkali deconstruction of sugarcane bagasse for improved hydrolysis: insight from small angle neutron scattering (SANS). Renew Energy 147:2091–2101

    Article  Google Scholar 

  • Hatzis C, Riley C, Philippidis G (1996) Detailed material balance and ethanol yield calculations for the biomass-to-ethanol conversion process. In: Seventeenth symposium on biotechnology for fuels and chemicals, Humana Press, Totowa, NJ, pp 443–459

  • Kim JS, Lee YY, Kim TH (2016) A review on alkaline pretreatment technology for bioconversion of lignocellulosic biomass. Bioresour Technol 199:42–48

    Article  CAS  PubMed  Google Scholar 

  • Ko JK, Bak JS, Jung MW, Lee HJ, Choi IG, Kim TH, Kim KH (2009) Ethanol production from rice straw using optimized aqueous-ammonia soaking pretreatment and simultaneous saccharification and fermentation processes. Bioresour Technol 100:4374–4380

    Article  CAS  PubMed  Google Scholar 

  • Lee HL, Chang CK, Teng KH, Liang PH (2011) Construction and characterization of different fusion proteins between cellulases and β-glucosidase to improve glucose production and thermostability. Bioresour Technol 102:3973–3976

    Article  CAS  PubMed  Google Scholar 

  • Liu Y, Zhang Y, Xu J, Sun Y, Yuan Z, Xie J (2015) Consolidated bioprocess for bioethanol production with alkali-pretreated sugarcane bagasse. Appl Energy 157:517–522

    Article  CAS  Google Scholar 

  • Liu CG, Xiao Y, Xia XX, Zhao XQ, Peng L, Srinophakun P, Bai FW (2019) Cellulosic ethanol production: progress, challenges and strategies for solutions. Biotechnol Adv 37:133–158

    Article  Google Scholar 

  • Marques NP, de Cassia PJ, Gomes E, da Silva R, Araujo AR, Ferreira H, Bocchini DA (2018) Cellulases and xylanases production by endophytic fungi by solid state fermentation using lignocellulosic substrates and enzymatic saccharification of pretreated sugarcane bagasse. Ind Crops Prod 122:66–75

    Article  CAS  Google Scholar 

  • McKee LS, Pena MJ, Rogowski A, Jackson A, Lewis RJ, York WS, Marles-Wright J (2012) Introducing endo-xylanase activity into an exo-acting arabinofuranosidase that targets side chains. Proc Natl Acad Sci USA 109:6537–6542

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mesa L, Gonzalez E, Cara C, Gonzalez M, Castro E, Mussatto SI (2011) The effect of organosolv pretreatment variables on enzymatic hydrolysis of sugarcane bagasse. Chem Eng J 168:1157–1162

    Article  CAS  Google Scholar 

  • Moura HO, Campos LM, da Silva VL, de Andrade JC, de Assumpçao SM, Pontes LA, de Carvalho LS (2018) Investigating acid/peroxide-alkali pretreatment of sugarcane bagasse to isolate high accessibility cellulose applied in acetylation reactions. Cellulose 25:5669–5685

    Article  CAS  Google Scholar 

  • Nath P, Dhillon A, Kumar K, Sharma K, Jamaldheen SB, Moholkar VS, Goyal A (2019) Development of bi-functional chimeric enzyme (CtGH1-L1-CtGH5-F194A) from endoglucanase (CtGH5) mutant F194A and β-1, 4-glucosidase (CtGH1) from Clostridium thermocellum with enhanced activity and structural integrity. Bioresour Technol 282:494–501

    Article  CAS  PubMed  Google Scholar 

  • Nedumaran M, Singh S, Jamaldheen SB, Nath P, Moholkar VS, Goyal A (2020) Assessment of combination of pretreatment of Sorghum durra stalk and production of chimeric enzyme (β-glucosidase and endo β-1, 4 glucanase, CtGH1-L1-CtGH5-F194A) and cellobiohydrolase (CtCBH5A) for saccharification to produce bioethanol. Prep Biochem Biotechnol 50:883–896

    Article  CAS  PubMed  Google Scholar 

  • Nelson N (1944) A photometric adaptation of the Somogyi method for the determination of glucose. Int J Biol Chem 153:375–380

    Article  CAS  Google Scholar 

  • Panagiotopoulos IA, Chandra RP, Saddler JN (2013) A two-stage pretreatment approach to maximise sugar yield and enhance reactive lignin recovery from poplar wood chips. Bioresour Technol 130:570–577

    Article  CAS  PubMed  Google Scholar 

  • Pandey A, Soccol CR, Nigam P, Soccol VT (2000) Biotechnological potential of agro-industrial residues. I: sugarcane bagasse. Bioresour Technol 74:69–80

    Article  CAS  Google Scholar 

  • Pathak KK, Das S (2020) Impact of bioenergy on environmental sustainability. Biomass valorization to bioenergy. Springer, Singapore, pp 133–158

    Chapter  Google Scholar 

  • Qiu Z, Aita GM, Walker MS (2012) Effect of ionic liquid pretreatment on the chemical composition, structure and enzymatic hydrolysis of energy cane bagasse. Bioresour Technol 117:251–256

    Article  CAS  PubMed  Google Scholar 

  • Raabo BE, Terkildsen TC (1960) On the enzymatic determination of blood glucose. Scand J Clin Lab Investig 12:402–407

    Article  CAS  Google Scholar 

  • Rabelo SC, Fonseca NA, Andrade RR, Maciel Filho R, Costa AC (2011) Ethanol production from enzymatic hydrolysis of sugarcane bagasse pretreated with lime and alkaline hydrogen peroxide. Biomass Bioenerg 35:2600–2607

    Article  CAS  Google Scholar 

  • Rezende CA, de Lima MA, Maziero P, deAzevedo ER, Garcia W, Polikarpov I (2011) Chemical and morphological characterization of sugarcane bagasse submitted to a delignification process for enhanced enzymatic digestibility. Biotechnol Biofuels 4:54

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rocha GJM, Andrade LP, Martin C, Araujo GT, Mouchrek Filho VE, Curvelo AAS (2020) Simultaneous obtaining of oxidized lignin and cellulosic pulp from steam-exploded sugarcane bagasse. Ind Crops Prod 147:112227–112232

    Article  CAS  Google Scholar 

  • Sant’Anna C, Souza W (2012) Microscopy as a tool to follow deconstruction of lignocellulosic biomass. In: Mendez-Vilas A (ed) Current microscopy contributions to advances in science and technology. Formatex Research Center, Badajoz, pp 639–645

    Google Scholar 

  • Satari B, Karimi K, Kumar R (2019) Cellulose solvent-based pretreatment for enhanced second-generation biofuel production: a review. Sustain Energy Fuels 3:11–62

    Article  CAS  Google Scholar 

  • Segal LGJMA, Creely JJ, Martin AE Jr, Conrad CM (1959) An empirical method for estimating the degree of crystallinity of native cellulose using the X-ray diffractometer. Text Res J 29:786–794

    Article  CAS  Google Scholar 

  • Sindhu R, Binod P, Janu KU, Sukumaran RK, Pandey A (2012) Organosolvent pretreatment and enzymatic hydrolysis of rice straw for the production of bioethanol. World J Microbiol Biotechnol 28:473–483

    Article  CAS  PubMed  Google Scholar 

  • Singh S, Gaikwad KK, Park SI, Lee YS (2017) Microwave-assisted step reduced extraction of seaweed (Gelidiella aceroso) cellulose nanocrystals. Int J Biol Macromol 99:506–510

    Article  CAS  PubMed  Google Scholar 

  • Somogyi M (1945) A new reagent for the determination of sugars. Int J Biol Chem 160:61–68

    Article  CAS  Google Scholar 

  • Sritrakul N, Nitisinprasert S, Keawsompong S (2017) Evaluation of dilute acid pretreatment for bioethanol fermentation from sugarcane bagasse pith. Agric Nat Resour 51:512–519

    Google Scholar 

  • Talha Z, Ding W, Mehryar E, Hassan M, Bi J (2016) Alkaline pretreatment of sugarcane bagasse and filter mud codigested to improve biomethane production. Biomed Res Int 2016:1–10

    Article  Google Scholar 

  • TAPPI (1992) Technical Association of Pulp and Paper Industry Atlanta Georgia, USA

  • Wang Q, Wang W, Tan X, Chen X, Guo Y, Yu Q, Zhuang X (2019) Low-temperature sodium hydroxide pretreatment for ethanol production from sugarcane bagasse without washing process. Bioresour Technol 291:121844–121850

    Article  CAS  PubMed  Google Scholar 

  • Xu F, Yu J, Tesso T, Dowell F, Wang D (2013) Qualitative and quantitative analysis of lignocellulosic biomass using infrared techniques: a mini-review. Appl Energy 104:801–809

    Article  CAS  Google Scholar 

  • Yan X, Cheng JR, Wang YT, Zhu MJ (2020) Enhanced lignin removal and enzymolysis efficiency of grass waste by hydrogen peroxide synergized dilute alkali pretreatment. Bioresour Technol 301:122756–1222764

    Article  CAS  PubMed  Google Scholar 

  • Zhang YHP, Cui J, Lynd LR, Kuang LR (2006) A transition from cellulose swelling to cellulose dissolution by o-phosphoric acid: evidence from enzymatic hydrolysis and supramolecular structure. Biomacromol 7:644–648

    Article  CAS  Google Scholar 

  • Zhang K, Pei Z, Wang D (2016) Organic solvent pretreatment of lignocellulosic biomass for biofuels and biochemicals: a review. Bioresour Technol 199:21–33

    Article  CAS  PubMed  Google Scholar 

  • Zhang H, Zhang J, Xie J, Qin Y (2020) Effects of NaOH-catalyzed organosolv pretreatment and surfactant on the sugar production from sugarcane bagasse. Bioresour Technol 312:123601–123607

    Article  CAS  PubMed  Google Scholar 

  • Zhao X, Cheng K, Liu D (2009) Organosolv pretreatment of lignocellulosic biomass for enzymatic hydrolysis. Appl Microbiol 82:815

    CAS  Google Scholar 

Download references

Acknowledgements

Authors acknowledge the financial support provided by DBT-Pan-IIT Grant (BT/EB/PAN-IIT 2012), Centre for Bioenergy from Department of Biotechnology, Ministry of Science and Technology, New Delhi, India to Prof. Arun Goyal. The fellowship to Priyanka Nath was supported by funding from DST Inspire fellowship from Department of Science and Technology, New Delhi, India.

Author information

Author notes

  1. Priyanka Nath and Premeshworii Devi Maibam have equally contributed to this work.

Authors and Affiliations

  1. Carbohydrate Enzyme Biotechnology Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India

    Priyanka Nath, Shweta Singh, Vikky Rajulapati & Arun Goyal

  2. Centre for Energy, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India

    Premeshworii Devi Maibam & Arun Goyal

  3. DBT PAN-IIT Center for Bioenergy, Indian Institute of Technology Guwahati, Guwahati, Assam, India

    Priyanka Nath, Shweta Singh & Arun Goyal

Authors
  1. Priyanka Nath
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Premeshworii Devi Maibam
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shweta Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vikky Rajulapati
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Arun Goyal
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

AG conceived the idea and designed the objectives. PN, MPD, and SS performed the experiments. VR performed FT-IR analysis. AG, PN, MPD, and SS wrote the paper.

Corresponding author

Correspondence to Arun Goyal.

Ethics declarations

Conflict of interest

There are no conflicts of interest to declare.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 91 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Nath, P., Maibam, P.D., Singh, S. et al. Sequential pretreatment of sugarcane bagasse by alkali and organosolv for improved delignification and cellulose saccharification by chimera and cellobiohydrolase for bioethanol production. 3 Biotech 11, 59 (2021). https://doi.org/10.1007/s13205-020-02600-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s13205-020-02600-y

Keywords

Search

Navigation