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3 Biotech

, 9:23 | Cite as

Benchmarking hydrolytic potential of cellulase cocktail obtained from mutant strain of Talaromyces verruculosus IIPC 324 with commercial biofuel enzymes

  • Lavika Jain
  • Akhilesh Kumar Kurmi
  • Deepti AgrawalEmail author
Original Article
  • 37 Downloads

Abstract

In the present study, an attempt was made to benchmark the hydrolytic potential of cellulase cocktail obtained from stable mutant UV-8 of Talaromyces verruculosus IIPC 324 (NFCCI 4117) with three commercially available cellulases. With two experimental approaches, acid-pretreated sugarcane bagasse was subjected to hydrolysis for 72 h, where all the enzymes were dosed on the basis of common protein or common cellulase activity /g cellulose content. Concentrated fungal enzyme (CFE) of mutant UV-8 resulted in ~ 59% and 55% saccharification of acid-pretreated sugarcane bagasse after 72 h at 55 °C and pH 4.5 with respect to reducing sugar release, when dosed at 25 mg protein/g and 500 IU CMC’ase/g cellulose, respectively. On the other hand, at similar dosages, the performance of Cellic CTec2 was best resulting in 77% and 66% saccharification, respectively. When enzyme desorption studies were undertaken by carrying out cellulase activities in saccharified broth after 72 h CFE of UV-8 emerged as the best cellulase cocktail. A minimum of 90% endoglucanase and 60% cellobiohydrolase I was successfully desorbed from residual biomass, thereby increasing the probability of enzyme recycle and reuse for next round of hydrolysis.

Keywords

Talaromyces verruculosus IIPC 324 Enzymatic saccharification Carboxy methyl cellulase (EG or CMC’ase) Reducing sugars (RS) CFE (concentrated fungal enzyme) Cellobiohydrolase (CBH) 

Notes

Acknowledgements

Authors are grateful to Dr Anjan Ray, Director CSIR-Indian Institute of Petroleum for providing necessary facilities to complete this work and constant encouragement. This research was funded by CSIR-IIP as in-house project under OLP- 350919. We would like to thank Dr Debashish Ghosh, Scientist Biofuel Division for kindly providing acid pretreated sugarcane bagasse for the entire study. Senior research fellowship awarded to Ms Lavika Jain by Council of Scientific and Industrial Research, New Delhi, India is greatly acknowledged.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

13205_2018_1547_MOESM1_ESM.docx (11 kb)
Supplementary material 1 (DOCX 11 KB)

References

  1. Application Sheet of Novozymes (2010) Cellic® CTec2 and HTec2—enzymes for hydrolysis of lignocellulosic materials. http://www.shinshu-u.ac.jp/faculty/engineering/chair/chem010/manual/Ctec2.pdf. Accessed on 4 July 2018
  2. Agrawal R, Gaur R, Mathur A, Kumar R, Gupta RP, Tuli DK, Satlewal A (2015) Improved saccharification of pilot-scale acid pretreated wheat straw by exploiting the synergistic behavior of lignocellulose degrading enzymes. RSC Adv 5:71462–71471CrossRefGoogle Scholar
  3. Anasontzis GE, Thuy NT, Hang DTM, Huong HT, Thanh DT, Hien DD, Thanh VN, Olsson L (2017) Rice straw hydrolysis using secretomes from novel fungal isolates from Vietnam. Biomass Bioenerg 99:11–20CrossRefGoogle Scholar
  4. Arantes V, Saddler JN (2011) Cellulose accessibility limits the effectiveness of minimum cellulase loading on the efficient hydrolysis of pretreated lignocellulosic substrates. Biotechnol Biofuels 4:3CrossRefGoogle Scholar
  5. Bissaro B, Røhr AK, Müller G, Chylenski P, Skaugen M, Forsberg Z, Horn SJ, Vaaje-Kolstad G, Eijsink VGH (2017) Oxidative cleavage of polysaccharides by monocopper enzymes depends on H2O2. Nat Chem Biol 13(10):1123–1128CrossRefGoogle Scholar
  6. 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 72:248–254CrossRefGoogle Scholar
  7. Cannella D, Hsieh CC, Felby C, Jørgensen H (2012) Production and effect of aldonic acids during enzymatic hydrolysis of lignocellulose at high dry matter content. Biotechnol Biofuels 5:26CrossRefGoogle Scholar
  8. De Castro AM, Carvalho MLDAD, Leite SGF, Pereira N Jr (2010) Cellulases from Penicillium funiculosum: production, properties and application to cellulose hydrolysis. J Indus Microbiol Biotechnol 37:151–158CrossRefGoogle Scholar
  9. Despande MV, Eriksson K, Pettersson LG (1984) An assay for selective determination of exo-1,4,-β-glucanases in a mixture of celluloytic enzymes. Anal Biochem 138:481–487CrossRefGoogle Scholar
  10. Ghosh TK (1987) Measurement of cellulase activities. Pure Appl Chem 59:257–268CrossRefGoogle Scholar
  11. Ghosh D, Dasgupta D, Agrawal D, Kaul S, Adhikari DK, Kurmi Ak, Arya PK, Bangwal D, Negi MS (2015) Fuels and chemicals from lignocellulosic biomass: an integrated biorefinery approach. Energ Fuel 29(5):3149–3157CrossRefGoogle Scholar
  12. Girard DJ, Conversa AO (1993) Recovery of cellulase from lignaceous hydrolysis residue. Appl Biochem Biotechnol 39–40:521–533CrossRefGoogle Scholar
  13. Harris PV, Welner D, McFarland KC, Re E, Polusen JN, Brown K, Salbo R, Ding H, Vlasenko E, Merino S, Xu F, Cherry J, Larsen S, Leggio LL (2010) Stimulation of lignocellulosic biomass hydrolysis by proteins of glycoside hydrolyase family 61: structure and function of a large, enigmatic family. Biochem 49:3305–3316CrossRefGoogle Scholar
  14. Jain L, Agrawal D (2018a) Rational approach for mutant selection of Talaromyces verruculosus IIPC 324 secreting biofuel cellulases-assessing saccharification potential. Ind Crops Prod 114:93–97.  https://doi.org/10.1016/j.indcrop.2018.01.078 CrossRefGoogle Scholar
  15. Jain L, Agrawal D (2018b) Performance evaluation of fungal cellulases with dilute acid pretreated sugarcane bagasse: A robust bioprospecting strategy for biofuel enzymes. Renewe Energ 115:978–988.  https://doi.org/10.1016/j.renene.2017.09.021 CrossRefGoogle Scholar
  16. Jérôme F, Chatel G, Vigier KDO (2016) Depolymerization of cellulose to processable glucans by non-thermal technologies. Green Chem 18: 3903–3391CrossRefGoogle Scholar
  17. Jung S, Song Y, Kim HM, Bae H (2015) Enhanced lignocellulosic biomass hydrolysis by oxidative lytic polysaccharide monooxygenase (LPMOs) GH61 from Gleophyllum trabeum. Enzyme Micro Technol 77:38–45CrossRefGoogle Scholar
  18. Klein-Marcuschamer D, Oleskowicz-Popiel P, Simmons BA, Blanch HW (2012) The challenge of enzyme cost in the production of lignocellulosic biofuels. Biotechnol Bioengg 109(4):1083–1087CrossRefGoogle Scholar
  19. Kovacs K, Macrelli S, Szakacs G, Zacchi G (2009) Enzymatic hydrolysis of steam-pretreated lignocellulosic materials with Trichoderma atroviride enzymes produced in-house. Biotechnol Biofuels 2:14CrossRefGoogle Scholar
  20. Kumar R, Wyman CE (2009) Effect of cellulase and xylanase enzymes on the deconstruction of solids from pretreatment of poplar by leading technologies. Biotechnol Prog 25(2):302–314CrossRefGoogle Scholar
  21. Li YL, Sun Z, Ge X, Zhang J (2016) Effect of lignin and surfactant on adsorption and hydrolysis of cellulases on cellulose. Biotechnol Biofuels 9:20CrossRefGoogle Scholar
  22. Lu X, Zheng X, Li X, Zhao J (2016) Adsorption and mechanism of cellulase enzymes onto lignin isolated from corn stover pretreated with liquid hot water. Biotechnol Biofuels 9:118CrossRefGoogle Scholar
  23. Mc Millan JD, Jenning EW, Mohagheghi A, Zuccarello M (2011) Comparative performance of pre-commercial cellulases hydrolysing pretreated corn stover. Biotechnol Biofuels 4:29CrossRefGoogle Scholar
  24. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31(3):426–428CrossRefGoogle Scholar
  25. Mohapatra S, Pattathil S, Thatoi H (2017) Structural and functional characterization of two Pennisetum sp. biomass during ultrasono-assisted alkali pretreatment and enzymatic hydrolysis for understanding the mechanism of targeted delignification and enhanced saccharification. ACS Sustain Chem Eng 5(8):6486–6497CrossRefGoogle Scholar
  26. Müller G, Várnai A, Johansen KS, Eijsink VGH, Horn SJ (2015) Harnessing the potential of LPMO-containing cellulase cocktails poses new demands on processing conditions. Biotechnol Biofuels 8:187CrossRefGoogle Scholar
  27. Pensupa N, Jin M, Kokolski M, Archer DB, Du C (2013) A solid state fungal fermentation-based strategy for the hydrolysis of wheat straw. Bioresour Technol 149:261–267CrossRefGoogle Scholar
  28. Pryor SW, Nahar N (2010) Deficiency of cellulase activity measurements for enzyme evaluation. Appl Biochem Biotechnol 162:1737–1750CrossRefGoogle Scholar
  29. Qi B, Chen X, Su Y, Wan Y (2011) Enzyme adsorption and recycling during hydrolysis of wheat straw lignocellulose. Bioresour Technol 102:2881–2889CrossRefGoogle Scholar
  30. Ramos LP, Silva L, Ballem AC, Pitarelo AP, Chiarello LM, Silveira MHL (2015) Enzymatic hydrolysis of steam-exploded sugarcane bagasse using high total solids and low enzyme loadings. Bioresour Technol 175:195–202CrossRefGoogle Scholar
  31. Reis L, Fontana RC, Delabona PS, Lima DJS, Camassola M, Pradella JGC, Dillon AJP (2013) Increased production of cellulases and xylanases by Penicillium echinulatum S1M29 in batch and fed-batch culture. Bioresour Technol 146:597–603CrossRefGoogle Scholar
  32. Rocha-Martín J, Martinez-Bernal C, Pérez-Cobas Y, Reyes-Sosa FM, García BD (2017) Additives enhancing enzymatic hydrolysis of lignocellulosic biomass. Bioresour Technol 244:48–56CrossRefGoogle Scholar
  33. Rodrigues AC, Haven MO, Lindedam J, Felby C, Gama M (2015) Celluclast and cellic® CTec2: saccharification/fermentation of wheat straw, solid–liquid partition and potential of enzyme recycling by alkaline washing. Enzyme Microb Technol 79–80:70–77CrossRefGoogle Scholar
  34. Sluiter A, Hames B, Ruiz R, Scarlata C, Sluiter J, Templeton D (2008) Determination of ash in biomass. Technical report. NREL/TP-510-42622Google Scholar
  35. Sluiter A, Hames B, Ruiz R, Scarlata C, Sluiter J, Templeton D, Crocker D (2012) Determination of structural carbohydrates and lignin in biomass laboratory. Technical report. NREL/TP-510-42618Google Scholar
  36. Sun FF, Hong J, Hu J, Saddler JN, Fang X, Zhang Z, Shen S (2015) Accessory enzymes influence cellulase hydrolysis of the model substrate and the realistic lignocellulosic biomass. Enzyme Microb Technol 79–80:42–48CrossRefGoogle Scholar
  37. Westereng B, Cannella D, Agger JW, Jørgensen H, Anderson ML, Eijsink VGH, Felby C (2015) Enzymatic cellulose oxidation is linked to lignin by long-range electron transfer. Sci Rep 5:18561.  https://doi.org/10.1038/srep18561 CrossRefPubMedPubMedCentralGoogle Scholar
  38. Yarbrough JM, Mittal A, Mansfield E, Taylor IILE, Hobdey SE, Sammond DW, Bomble YJ, Crowley MF, Decker SR, Himmel ME, Vinzant TB (2015) New perspective on glycoside hydrolase binding to lignin from pretreated corn stover. Biotechnol Biofuels 8:214CrossRefGoogle Scholar
  39. Zhu Z, Sathitsuksanoh N, Zhang PYH (2009) Direct quantitative determination of adsorbed cellulase on lignocellulosic biomass with its application to study cellulase desorption for potential recycling. Analyst 134:2267–2272CrossRefGoogle Scholar

Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  1. 1.Biotechnology Conversion Area, Biofuels DivisionCSIR-Indian Institute of PetroleumDehradunIndia
  2. 2.Academy of Scientific and Innovative Research (AcSIR), CSIR-HRDC CampusGhaziabadIndia

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