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Characterization of magnetic nanoparticle–immobilized cellulases for enzymatic saccharification of rice straw

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Abstract

Cellulases convert lignocellulosic biomass into fermentable sugars which further act as substrate for ethanol production. Our previous research was focused on enzymatic hydrolysis of rice straw by free cellulases for production of fermentable sugars. Immobilization is a powerful tool to increase the stability and reusability of cellulases besides improving the economy of ethanol production process from rice straw. In the present study, cellulase produced from Aspergillus fumigatus was immobilized on magnetic nanoparticles by using glutaraldehyde cross linker with a binding efficiency of 65.55%. The electron microscopy and spectroscopy tools confirmed the enzyme immobilization process on magnetic nanoparticles. The immobilized cellulase exhibited filter paper, carboxymethyl cellulase and cellobiase activities of 11.82, 21.36 and 10.81 IU, respectively. The free and immobilized cellulase exhibited identical pH optima (pH 5.0) while different temperature optima of 50 °C and 60 °C, respectively. The immobilized enzyme retained 56.87% of its maximal activity after 6 h of pre-incubation at 60 °C. Km (Michaelis constant) and Vmax (maximum velocity) of immobilized enzyme were 11.76 mM and 1.17 μmol min−1 ml−1, respectively. The immobilized cellulase hydrolysed pre-treated rice straw with saccharification efficiency of 52.67%. Further, it could be reutilized for up to four saccharification cycles with retention of 50.34% activity. Therefore, the improved properties of magnetic nanoparticle-immobilized cellulase and its reusability benefits offer a promising potential for industrial production of fermentable sugars and ethanol from rice straw.

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References

  1. Wyman CE, Cai CM, Kumar R (2019) Bioethanol from lignocellulosic biomass. In: Kaltschmitt M (ed) Energy from organic materials (biomass) a volume in the encyclopedia of sustainability science and technology, 2nd edn. Springer, New York, pp 997–1022

    Google Scholar 

  2. Zhang Z, Liu B, Zhao ZK (2012) Efficient acid catalyzed hydrolysis of cellulose in organic electrolyte solutions. Polym Degrad Stab 97:573–577

    Article  Google Scholar 

  3. Kuila A, Sharma V, Garlapati VK, Singh A, Roy L, Banerjee R (2016) Present statu s on enzymatic hydrolysis of lignocellulosic biomass for bioethanol production. Adv Biofeedstocks Biofuels 1:85

    Google Scholar 

  4. Brummer V, Jurena T, Hlavacek V, Omelkova J, Bebar L, Gabriel P, Stehlik P (2014) Enzymatic hydrolysis of pretreated waste paper–source of raw material for production of liquid biofuels. Bioresour Technol 152:543–547

    Article  Google Scholar 

  5. Madadi M, Tu Y, Abbas A (2017) Recent status on enzymatic saccharification of lignocellulosic biomass for bioethanol production. Electron J Biol 13:135–143

    Google Scholar 

  6. Ahamed A, Vermette P (2008) Culture based strategies to enhance cellulase enzyme production from Trichoderma reesei RUT-30 in bioreactor culture conditions. Biochem Eng 140:399–407

    Article  Google Scholar 

  7. Zhang Q, Han X, Tang B (2013) Preparation of a magnetically recoverable biocatalyst support on monodisperse Fe3O4 nanoparticles. RSC Adv 3:9924–9931

    Article  Google Scholar 

  8. Chundawat S, Sousa LDC, Cheh AM, Balan V, Dale B (2017) Methods for producing extracted and digested products from pretreated lignocellulosic biomass. U.S. patent, 9650657

  9. Mitchell DT, Lee SB, Trofin L, Li N, Nevanen TK, Soderlund H, Martin CR (2002) Smart nanotubes for bioseparations and biocatalysis. J Am Chem Soc 124:11864–11865

    Article  Google Scholar 

  10. Eldin MSM (2016) Enzyme immobilization: nanopolymers for enzyme immobilization applications. Energy 16:18

    Google Scholar 

  11. Huang WC, Wang W, Xue C, Mao X (2018) Effective enzyme immobilization onto a magnetic chitin nanofiber composite. ACS Sustain Chem Eng 6:8118–8124

    Article  Google Scholar 

  12. Ahmad R, Khare SK (2018) Immobilization of Aspergillus niger cellulase on multiwall carbon nanotubes for cellulose hydrolysis. Bioresour Technol 252:72–75

    Article  Google Scholar 

  13. Otari SV, Patel SK, Kim SY, Haw JR, Kalia VC, Kim IW, Lee JK (2019) Copper ferrite magnetic nanoparticles for the immobilization of enzyme. Indian J Microbiol 59:105–108

    Article  Google Scholar 

  14. Abbaszadeh M, Hejazi P (2019) Metal affinity immobilization of cellulase on Fe3O4 nanoparticles with copper as ligand for biocatalytic applications. Food Chem 290:47–55

    Article  Google Scholar 

  15. Verma ML, Barrow CJ, Puri M (2013) Nanobiotechnology as a novel paradigm for enzyme immobilisation and stabilisation with potential applications in biodiesel production. Appl Microbiol Biotechnol 97:23–39

    Article  Google Scholar 

  16. Kim K, Lee OK, Lee E (2018) Nano-immobilized biocatalysts for biodiesel production from renewable and sustainable resources. J Catal 8:68

    Google Scholar 

  17. García PF, Brammen M, Wolf M, Reinlein S, von Roman MF, Berensmeier S (2015) High-gradient magnetic separation for technical scale protein recovery using low cost magnetic nanoparticles. Sep Purif Technol 150:29–36

    Article  Google Scholar 

  18. Kumar A, Singh S, Nain L (2018) Magnetic nanoparticle immobilized cellulase enzyme for saccharification of paddy straw. Int J Curr Microbiol App Sci 7:881–893

    Article  Google Scholar 

  19. Abraham RE, Verma ML, Barrow CJ, Puri M (2014) Suitability of magnetic nanoparticle immobilised cellulases in enhancing enzymatic saccharification of pretreated hemp biomass. Biotechnol Biofuels 7:90

    Article  Google Scholar 

  20. Selvam K, Govarthanan M, Senbagam D, Kamala-Kannan S, Senthilkumar B, Selvankumar T (2016) Activity and stability of bacterial cellulase immobilized on magnetic nanoparticles. Chin J Catal 37:1891–1898

    Article  Google Scholar 

  21. **ao A, Xu C, Lin Y, Ni H, Zhu Y, Cai H (2016) Preparation and characterization of κ-carrageenase immobilized onto magnetic iron oxide nanoparticles. Electron J Biotechnol 19:1–7

    Article  Google Scholar 

  22. Han J, Luo P, Wang Y, Wang L, Li C, Zhang W, Dong J, Ni L (2018) The development of nanobiocatalysis via the immobilization of cellulase on composite magnetic nanomaterial for enhanced loading capacity and catalytic activity. Int J Biol Macromol 119:692–700

    Article  Google Scholar 

  23. Dong RJ, Zheng DF, Yang DJ, Qiu XQ (2019) pH-responsive lignin-based magnetic nanoparticles for recovery of cellulase. Bioresour Technol 294:122133

    Article  Google Scholar 

  24. Kumar A, Singh S, Tiwari R, Goel R, Nain L (2017) Immobilization of indigenous holocellulase on iron oxide (Fe2O3) nanoparticles enhanced hydrolysis of alkali pretreated paddy straw. Int J Biol Macromol 96:538–549

    Article  Google Scholar 

  25. Zhang Q, Kang J, Yang B, Zhao L, Hou Z, Tang B (2016) Immobilized cellulase on Fe3O4 nanoparticles as a magnetically recoverable biocatalyst for the decomposition of corncob. Chin J Catal 37:389–397

    Article  Google Scholar 

  26. Manasa P, Saroj P, Korrapati N (2017) Immobilization of cellulase enzyme on zinc ferrite nanoparticles in increasing enzymatic hydrolysis on ultrasound assisted alkaline pretreated Crotalaria juncea biomass. Indian J Sci Technol 10:1–7

    Article  Google Scholar 

  27. Poorakbar E, Shafiee A, Saboury AA, Rad BL, Khoshnevisan K, Ma'mani L, Derakhshankhah H, Ganjali MR, Hosseini M (2018) Synthesis of magnetic gold mesoporous silica nanoparticles core shell for cellulase enzyme immobilization: improvement of enzymatic activity and thermal stability. Process Biochem 71:92–100

    Article  Google Scholar 

  28. 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 101:4767–4774

    Article  Google Scholar 

  29. Raj T, Kapoor M, Gaur R, Christopher J, Lamba BY, Tuli DK, Kumar R (2015) Physical and chemical characterization of various Indian agriculture residues for biofuels production. Energ Fuels 29:3111–3118

    Article  Google Scholar 

  30. Colonia BSO, Junior AFC (2014) Screening and detection of extracellular cellulases (endo-and exo-glucanases) secreted by filamentous fungi isolated from soils using rapid tests with chromogenic dyes. Afr J Biotechnol 13:4694–4701

    Article  Google Scholar 

  31. Meyyappan A, Shakila BA, Kurian GA (2015) One step synthesis of iron oxide nanoparticles via chemical and green route- an effective comparison. Int J Pharm Pharm Sci 7:70–74

    Google Scholar 

  32. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Article  Google Scholar 

  33. Mandels M, Andreotti R, Roche C (1976) Measurement of saccharifying cellulase. Biotechnol Bioeng Symp. Pp 21-33. US Army Natick development center, Nauck, MA

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

    Article  Google Scholar 

  35. Wood TM, Bhat KM (1988) Methods for measuring cellulase activities. Methods Enzymol 160:87–112

    Article  Google Scholar 

  36. Toyama M, Ogawa K (1977) Cellulase production of Trichoderma viride in solid and submerged culture methods. In: Ghose TK (ed) Proc bioconversion of cellulosic substances into energy, chemicals and microbial protein, vol 1. IIT, New Delhi, pp 305–327

    Google Scholar 

  37. Xu J, Huo S, Yuan Z, Zhang Y, Xu H, Guo Y, Liang C, Zhuang X (2011) Characterization of direct cellulase immobilization with superparamagnetic nanoparticles. Biocatal Biotransform 29:71–76

    Article  Google Scholar 

  38. Lu PJ, Fu WE, Huang SC, Lin CY, Ho ML, Chen YP, Cheng HF (2018) Methodology for sample preparation and size measurement of commercial ZnO nanoparticles. J Food Drug Anal 26:628–636

    Article  Google Scholar 

  39. Predoi D (2007) A study on iron oxide nanoparticles coated with dextrin obtained by coprecipitation. Dig J Nanomater Bios 2:169–173

    Google Scholar 

  40. Burgula Y, Khali D, Kim S, Krishnan SS, Cousin MA, Gore JP, Reuhs BL, Mauer LJ (2006) Detection of Escherichia coli O157:H7 and Salmonella typhimurium using filtration followed by Fourier-transform infrared spectroscopy. J Food Prot 69:1777–1784

    Article  Google Scholar 

  41. Kaur P, Kocher GS, Taggar MS, Sooch SS, Kumar V (2017) Assessment of thermophilic fungal strains for cellulase production using chemical pretreated rice straw. Chem Sci Rev Lett 6:629–634

    Google Scholar 

  42. Crompton EW, Maynard LA (1938) The relation of cellulose and lignin content to the nutritive value of animal feeds. J Anim Nut 15:391–392

    Google Scholar 

  43. Goering HK, Van Soest PJ (1970) Forage fibre analysis. Pp. 1-20. Agricultural research services, Washington DC, U.S.

  44. AOAC (2000) Official methods of analysis, 17th edn. Association of Official Analytical Chemists, Washington, D.C.

    Google Scholar 

  45. Singh A, Tuteja S, Singh N, Bishnoi NR (2011) Enhanced saccharification of rice straw and hull by microwave-alkali pre-treatment and lignocellulolytic enzyme production. Bioresour Technol 102:1773–1782

    Article  Google Scholar 

  46. Jia J, Zhang W, Yang Z, Yang X, Wang N, Yu X (2017) Novel magnetic cross-linked cellulase aggregates with a potential application in lignocellulosic biomass bioconversion. Molecules 22:269

    Article  Google Scholar 

  47. Mohamed SA, Al-Harbi MH, Almulaiky YQ, Ibrahim IH, El-Shishtawy RM (2017) Immobilization of horseradish peroxidase on Fe3O4 magnetic nanoparticles. Electron J Biotechnol 27:84–90

    Article  Google Scholar 

  48. Kouassi GK, Irudayaraj J, McCarty G (2005) Examination of cholesterol oxidase attachment to magnetic nanoparticles. J Nanobiotechnol 3:1

    Article  Google Scholar 

  49. Can HK, Kavlak S, Parvizi Khosroshahi S, Güner A (2018) Preparation, characterization and dynamical mechanical properties of dextran-coated iron oxide nanoparticles (DIONPs). Artif Cells Nanomed Biotechnol 46:421–431

    Article  Google Scholar 

  50. Chittur KK (1998) FTIR/ATR for protein adsorption to biomaterial surfaces. Biomater 19:357–369

    Article  Google Scholar 

  51. Herzberg G, Crawford BL Jr (1946) Infrared and Raman spectra of polyatomic molecules. J Phys Chem 50:288

    Article  Google Scholar 

  52. Jordan J, Kumar CS, Theegala C (2011) Preparation and characterization of cellulase-bound magnetite nanoparticles. J Mol Catal B Enzym 68:139–146

    Article  Google Scholar 

  53. Khoshnevisan K, Bordbar AK, Zare D, Davoodi D, Noruzi M, Barkhi M, Tabatabaei M (2011) Immobilization of cellulase enzyme on superparamagnetic nanoparticles and determination of its activity and stability. Chem Eng J 171:669–673

    Article  Google Scholar 

  54. Tao QL, Li Y, Shi Y, Liu RJ, Zhang YW, Guo J (2016) Application of molecular imprinted magnetic Fe3O4@SiO2 nanoparticles for selective immobilization of cellulase. J Nanosci Nanotechnol 16:6055–6060

    Article  Google Scholar 

  55. Tu M, Zhang X, Kurabi A, Gilkes N, Mabee W, Saddler J (2006) Immobilization of β-glucosidase on Eupergit C for lignocellulose hydrolysis. Biotechnol Lett 28:151–156

    Article  Google Scholar 

  56. Weetall HH (1993) Preparation of immobilized proteins covalently coupled through silane coupling agents to inorganic supports. Appl Biochem Biotechnol 41:157–188

    Article  Google Scholar 

  57. Brodeur G, Yau E, Badal K, Collier J, Ramachandran KB, Ramakrishnan S (2011) Chemical and physiochemical pretreatment of lignocellulosic biomass: a review. Enzym Res 2011:782532

    Article  Google Scholar 

  58. Hendriks ATWM, Zeeman G (2009) Pretreatments to enhance the digestibility of lignocellulosic biomass. Bioresour Technol 100:10–18

    Article  Google Scholar 

  59. Weerasai K, Suriyachai N, Poonsrisawat A, Arnthong J, Unrean P, Laosiripojana N, Champreda V (2014) Sequential acid and alkaline pretreatment of rice straw for bioethanol fermentation. BioResources 9:5988–6001

    Article  Google Scholar 

  60. Syazwanee MGMF, Shaziera AN, Izzati MZNA, Azwady AN, Muskhazli A (2018) Improvement of delignification, desilication and cellulosic content availability in paddy straw via physico-chemical pretreatments. Annu Res Rev Biol 6:1–11

    Article  Google Scholar 

  61. Huang PJ, Chang KL, Hsieh JF, Chen ST (2015) Catalysis of rice straw hydrolysis by the combination of immobilized cellulase from Aspergillus niger on β-cyclodextrin-Fe3O4 nanoparticles and ionic liquid. Biomed Res Int 2015:409103

    Google Scholar 

  62. Baskar G, Kumar RN, Melvin XH, Aiswarya R, Soumya S (2016) Sesbania aculeate biomass hydrolysis using magnetic nanobiocomposite of cellulase for bioethanol production. Renew Energy 98:23–28

    Article  Google Scholar 

  63. Periyasamy K, Santhalembi L, Mortha G, Aurousseau M, Boyer A, Subramanian S (2018) Bioconversion of lignocellulosic biomass to fermentable sugars by immobilized magnetic cellulolytic enzyme cocktails. Langmuir 34:6546–6555

    Article  Google Scholar 

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Acknowledgements

Authors are thankful to Department of Renewable Energy Engineering, Punjab Agricultural University, Ludhiana, Punjab, India, for providing the necessary facilities to undertake this study. Authors are thankful to Sophisticated Analytical Instrumentation Facility, Panjab University, Chandigarh, Punjab, India, for providing instrumentation for analysis of sample by SEM, EDS and XRD.

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  1. Department of Biochemistry, Punjab Agricultural University, Ludhiana, 141004, India

    Prabhpreet Kaur

  2. Department of Renewable Energy Engineering, Punjab Agricultural University, Ludhiana, 141004, India

    Monica Sachdeva Taggar

  3. Electron Microscopy and Nanoscience Laboratory, Department of Soil Science, Punjab Agricultural University, Ludhiana, 141004, India

    Anu Kalia

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  1. Prabhpreet Kaur
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Kaur, P., Taggar, M.S. & Kalia, A. Characterization of magnetic nanoparticle–immobilized cellulases for enzymatic saccharification of rice straw. Biomass Conv. Bioref. 11, 955–969 (2021). https://doi.org/10.1007/s13399-020-00628-x

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