A novel approach to produce glucose from the supernatant obtained upon the dilute acid pre-treatment of rice straw and synergistic action of hydrolytic enzymes producing microbes

  • Manisha Chownk
  • Rajender Singh Sangwan
  • Sudesh Kumar YadavEmail author
Biotechnology and Industrial Microbiology - Research Paper


The present work refers to a process involving the use of dilute nitric acid pretreatment and enzymatic hydrolysis for the transformation of rice straw into simple sugars. Acid pre-treated rice straw was separated into the pulp and supernatant through centrifugation and filtration. The two fractions are then converted into simple sugars by combined action of microbes producing cellulase and laccase enzymes. These microbes were isolated from soil samples which were collected from different locations with varying altitudes, expected to harbour microbes with high-hydrolysing activity. The nitric acid pretreatment was carried out at 30 °C, 200 rpm for 72 h. After 72 h, the culture supernatants were analysed for the presence of glucose with the help of HPLC. The supernatant fraction separated after the acid pre-treated rice straw produced highest amount of glucose (205 mg/g of rice straw) upon subsequent hydrolysis with synergistic action of cellulase and laccase-producing microbes.


Rice straw Cellulose Laccase Glucose Biomass transformation 



Authors are thankful to Mr. Umesh Singh, STA, CIAB, for his help in analysis.

Funding information

Authors are thankful to the Department of Biotechnology (DBT), GOI for the financial support to conduct this research.

Supplementary material

42770_2018_13_MOESM1_ESM.docx (119 kb)
ESM 1 (DOCX 118 kb)


  1. 1.
    Sangakkara UR (2005) Rice almanac–source book for the most important economic activity on Earth, eds Maclean JL. Dawe DC. Hardy B. Hettel GP. Wallingford: CABI. The Journal of Agricultural Science 143(1):110Google Scholar
  2. 2.
    Adhikari A, Sekhon MK (2014) Export of Basmati rice from India: performance and trade direction. J Agric Dev Policy 24(1):1–13 ISSN: 2322-0457Google Scholar
  3. 3.
    Rahnama N, Foo HL, Abdul Rahman NA, Ariff A, Md Shah UK (2014) Saccharification of rice straw by cellulase from a local Trichoderma harzianum SNRS3 for biobutanol production. BMC Biotechnol 14(1):103. CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Gadde B, Bonnet S, Menke C, Garivait S (2009) Air pollutant emissions from rice straw open field burning in India, Thailand and the Philippines. Environ. Pollution 157(5):1554–1558. CrossRefGoogle Scholar
  5. 5.
    Diep NQ, Sakanishi K (2011) Potential of bioethanol production from agricultural residues in the Mekong delta, Vietnam. Int Energy J 12:145–154. CrossRefGoogle Scholar
  6. 6.
    Hadar Y (2013) Sources for lignocellulosic raw materials for the production of ethanol. In Lignocellulose Conversion: 21-38, Springer Berlin Heidelberg doi: CrossRefGoogle Scholar
  7. 7.
    Kopke M, Noack S, Dürre P (2011) The Past, present, and future of biofuels – biobutanol as promising alternative, biofuel production-recent developments and prospects. In: Bernards MADS (ed) Biofuel production-recent developments and prospects. InTech, London, pp 451–486. CrossRefGoogle Scholar
  8. 8.
    Brodeur G, Yau E, Badal K, Collier J, Ramachandran KB, Ramakrishnan S (2011) Chemical and physicochemical pretreatment of lignocellulosic biomass: a review. Enzym Res 787532:1–17. CrossRefGoogle Scholar
  9. 9.
    Mosier N, Wyman C, Dale B, Elander R, Lee YY, Holtzapple M, Ladisch M (2005) Features of promising technologies for pretreatment of lignocellulosic biomass. Bioresour Technol 96(6):673–686. CrossRefGoogle Scholar
  10. 10.
    Holladay J, Bozell J, White J, Johnson D (2007) Top value-added chemicals from biomass. DOE Report PNNL-16983.
  11. 11.
    Kumar V, Satyanarayana T (2011) Applicability of thermo-alkali-stable and cellulase-free xylanase from a novel thermo-halo-alkaliphilic Bacillus halodurans in producing xylooligosaccharides. Biotechnol Lett 33(11):2279–2285. CrossRefPubMedGoogle Scholar
  12. 12.
    Wi SG, Choi IS, Kim KH, Kim HM, Bae HJ (2013) Bioethanol production from rice straw by popping pretreatment. Biotechnol Biofuels 6(1):166. CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Hashem M, Ali EH, Abdel-Basset R (2013) Recycling rice straw into biofuel “ethanol” by Saccharomyces cerevisiae and Pichia guilliermondii. J Agri Sci Technol 15(4):709–721 ISSN:16807073Google Scholar
  14. 14.
    Song HT, Gao Y, Yang YM, Xiao WJ, Liu SH, Xia WC, Liu ZL, Yi L, Jiang ZB (2016) Synergistic effect of cellulase and xylanase during hydrolysis of natural lignocellulosic substrates. Biores Technol 219:710–715. CrossRefGoogle Scholar
  15. 15.
    Nakatani Y, Yamada R, Ogino C, Kondo A (2013) Synergetic effect of yeast cell-surface expression of cellulase and expansin-like protein on direct ethanol production from cellulose. Microb Cell Factories 12(1):66. CrossRefGoogle Scholar
  16. 16.
    Nirmala P, Sindhu A (2011) Production of endoglucanase by optimizing the environmental conditions of Bacillus circulans on submerged fermentation. Int J Appl Eng Res Dindigul 2:472–481 ISSN - 0976-4259Google Scholar
  17. 17.
    Sheikhi F, Ardakani MR, Enayatizamir N, Rodriguez-Couto S (2012) The determination of assay for laccase of Bacillus subtilis WPI with two classes of chemical compounds as substrates. Indian J Microbiol 52(4):701–707. CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    O’Sullivan C, Christopher, Busby BB, Mizrachi IK (2017) Managing Sequence Data. In: Keith JM (ed) Bioinformatics: volume I: data, sequence analysis, and evolution. Springer Publishing, New York, pp 79–106Google Scholar
  19. 19.
    McGinnis S, Madden TL (2004) BLAST: at the core of a powerful and diverse set of sequence analysis tools. Nucleic Acids Res 32(Web Server):W20–W25. CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Benson DA, Cavanaugh M, Clark K, Karsch-Mizrachi I, Lipman DJ, Ostell J, Sayers EW (2013) GenBank. Nucleic Acids Res 41(Database issue):D36–D42. CrossRefPubMedGoogle Scholar
  21. 21.
    Harmsen PFH, Huijgen W, Bermudez L, Bakker R (2010) Literature review of physical and chemical pretreatment processes for lignocellulosic biomass. Wageningen UR Food & Biobased Research, Wageningen, p 1184Google Scholar
  22. 22.
    Binod P, Sindhu R, Singhania RR, Vikram S, Devi L, Nagalakshmi S, Kurien N, Sukumaran RK, Pandey A (2010) Bioethanol production from rice straw: an overview. Bioresour Technol 101(13):4767–4774. CrossRefPubMedGoogle Scholar
  23. 23.
    Aderemi BO, Abu E, Highina BK (2008) The kinetics of glucose production from rice straw by Aspergillus niger. Afr J Biotechnol 7(11):1745–1752. CrossRefGoogle Scholar
  24. 24.
    Kshirsagar SD, Waghmare PR, Chandrakant Loni P, Patil SA, Govindwar SP (2015) Dilute acid pretreatment of rice straw, structural characterization and optimization of enzymatic hydrolysis conditions by response surface methodology. RSC Adv 5(58):46525–46533. CrossRefGoogle Scholar
  25. 25.
    Lu X, Zhang Y, Angelidaki I (2009) Optimization of H2SO4-catalyzed hydrothermal pretreatment of rapeseed straw for bioconversion to ethanol: focusing on pretreatment at high solids content. Bioresour Technol 100(12):3048–3053. CrossRefPubMedGoogle Scholar
  26. 26.
    Drapcho CM, Nhuan NP, Walker TH (2008) Biofuels engineering process technology. McGraw-Hill, New YorkGoogle Scholar
  27. 27.
    Ogunyewo OA, Olajuyigbe FM (2016) Unravelling the interactions between hydrolytic and oxidative enzymes in degradation of lignocellulosic biomass by Sporothrix carnis under various fermentation conditions. Biochem Res Int 2016:1–8. CrossRefGoogle Scholar
  28. 28.
    Saritha M, Arora A, Lata (2012) Biological pretreatment of lignocellulosic substrates for enhanced delignification and enzymatic digestibility. Indian J Microbiol 52(2):122–130. CrossRefPubMedGoogle Scholar
  29. 29.
    Farone WA, Cuzens JE (1996) Method of producing sugars using strong acid hydrolysis of cellulosic and hemicellulosic materials. U.S. Patent 5562777Google Scholar
  30. 30.
    Yoon KP (2010) Method for pretreating biomass to produce bioethanol. U.S patent 2010131829 A1Google Scholar
  31. 31.
    Varanasi S, Schall CA, Dadi AP, et al (2011) Biomass pretreatment. U.S. Patent 8030030Google Scholar
  32. 32.
    Gordon RE, Haynes WC, Pang CHN (1973) The genus Bacillus. In: O’Leary WM (ed) Practical handbook of microbiology. CRC Press, Boca Raton, pp 109–126Google Scholar
  33. 33.
    Wang H, Xiang T, Wang Y, Song J, Zhai Y, Chen X, Li Y, Zhao B, Zhao B, Ruan Z (2014) Microbacterium petrolearium sp. nov., isolated from an oil-contaminated water sample. Int J Syst Evol Microbiol 64(Pt 12):4168–4172. CrossRefPubMedGoogle Scholar
  34. 34.
    Niu K, Chen P, Zhang X, Tan W-S (2009) Enhanced enzymatic hydrolysis of rice straw pretreated by alkali assisted with photocatalysis technology. J Chem Technol Biotechnol 84(8):1240–1245. CrossRefGoogle Scholar
  35. 35.
    Poornejad N, Karimi K, Behzad T (2014) Ionic liquid pretreatment of rice straw to enhance saccharification and bioethanol production. J Biomass Biofuel.

Copyright information

© Sociedade Brasileira de Microbiologia 2018

Authors and Affiliations

  • Manisha Chownk
    • 1
  • Rajender Singh Sangwan
    • 1
  • Sudesh Kumar Yadav
    • 1
    Email author
  1. 1.Biotechnology and Synthetic BiologyCenter of Innovative and Applied Bioprocessing (CIAB)MohaliIndia

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