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

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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.

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References

  • 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

  • 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 5:71462–71471

    Article  CAS  Google Scholar 

  • 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 99:11–20

    Article  CAS  Google Scholar 

  • Arantes V, Saddler JN (2011) Cellulose accessibility limits the effectiveness of minimum cellulase loading on the efficient hydrolysis of pretreated lignocellulosic substrates. Biotechnol Biofuels 4:3

    Article  CAS  Google Scholar 

  • 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):1123–1128

    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  Google Scholar 

  • 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 5:26

    Article  CAS  Google Scholar 

  • 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 37:151–158

    Article  CAS  Google Scholar 

  • 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 138:481–487

    Article  Google Scholar 

  • Ghosh TK (1987) Measurement of cellulase activities. Pure Appl Chem 59:257–268

    Article  Google Scholar 

  • 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):3149–3157

    Article  CAS  Google Scholar 

  • Girard DJ, Conversa AO (1993) Recovery of cellulase from lignaceous hydrolysis residue. Appl Biochem Biotechnol 39–40:521–533

    Article  Google Scholar 

  • 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 49:3305–3316

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • Jérôme F, Chatel G, Vigier KDO (2016) Depolymerization of cellulose to processable glucans by non-thermal technologies. Green Chem 18: 3903–3391

    Article  Google Scholar 

  • 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 77:38–45

    Article  CAS  Google Scholar 

  • 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):1083–1087

    Article  CAS  Google Scholar 

  • 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 2:14

    Article  Google Scholar 

  • 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):302–314

    Article  CAS  Google Scholar 

  • Li YL, Sun Z, Ge X, Zhang J (2016) Effect of lignin and surfactant on adsorption and hydrolysis of cellulases on cellulose. Biotechnol Biofuels 9:20

    Article  Google Scholar 

  • 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 9:118

    Article  Google Scholar 

  • Mc Millan JD, Jenning EW, Mohagheghi A, Zuccarello M (2011) Comparative performance of pre-commercial cellulases hydrolysing pretreated corn stover. Biotechnol Biofuels 4:29

    Article  CAS  Google Scholar 

  • Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31(3):426–428

    Article  CAS  Google Scholar 

  • 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):6486–6497

    Article  CAS  Google Scholar 

  • 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 8:187

    Article  Google Scholar 

  • 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 149:261–267

    Article  CAS  Google Scholar 

  • Pryor SW, Nahar N (2010) Deficiency of cellulase activity measurements for enzyme evaluation. Appl Biochem Biotechnol 162:1737–1750

    Article  CAS  Google Scholar 

  • Qi B, Chen X, Su Y, Wan Y (2011) Enzyme adsorption and recycling during hydrolysis of wheat straw lignocellulose. Bioresour Technol 102:2881–2889

    Article  CAS  Google Scholar 

  • 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 175:195–202

    Article  CAS  Google Scholar 

  • 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 146:597–603

    Article  Google Scholar 

  • Rocha-Martín J, Martinez-Bernal C, Pérez-Cobas Y, Reyes-Sosa FM, García BD (2017) Additives enhancing enzymatic hydrolysis of lignocellulosic biomass. Bioresour Technol 244:48–56

    Article  Google Scholar 

  • 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 79–80:70–77

    Article  Google Scholar 

  • Sluiter A, Hames B, Ruiz R, Scarlata C, Sluiter J, Templeton D (2008) Determination of ash in biomass. Technical report. NREL/TP-510-42622

  • 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-510-42618

  • 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 79–80:42–48

    Article  Google Scholar 

  • 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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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 8:214

    Article  Google Scholar 

  • 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 134:2267–2272

    Article  CAS  Google Scholar 

Download references

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.

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Correspondence to Deepti Agrawal.

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Jain, L., Kurmi, A.K. & Agrawal, D. Benchmarking hydrolytic potential of cellulase cocktail obtained from mutant strain of Talaromyces verruculosus IIPC 324 with commercial biofuel enzymes. 3 Biotech 9, 23 (2019). https://doi.org/10.1007/s13205-018-1547-x

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