Effective Substrate Loading for Saccharification of Corn Cob and Concurrent Production of Lignocellulolytic Enzymes by Fusarium oxysporum and Sporothrix carnis

Author(s): Folasade M. Olajuyigbe*, Cornelius O. Fatokun, Oluwatosin I. Oni

Journal Name: Current Biotechnology

Volume 8 , Issue 2 , 2019

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Graphical Abstract:


Background: One of the critical challenges of cost-effective bioethanol production from lignocellulosic biomass is the decreasing yield of reducing sugars caused by increasing substrate loading. Hence, it is crucial to determine the best substrate concentration for efficient saccharification of lignocellulosic wastes.

Objective: This paper reports the saccharification of corn cob by two lignocellulolytic fungi (Fusarium oxysporum and Sporothrix carnis) and concurrent production of lignocellulolytic enzymes at varying substrate concentrations.

Methods: F. oxysporum and S. carnis were cultivated on corn cob based media at 30°C and 160 rpm for 144 h. The lignocellulosic composition of corn cob was determined. Saccharification of varying concentrations of substrate was determined by evaluating the release of reducing sugar while the production of cellulase and xylanase was monitored.

Results: Cellulose, hemicellulose and lignin contents of corn cob were 37.8±1.56%, 42.2±1.68% and 12.7±1.23%, respectively. Yields of reducing sugar by F. oxysporum and S. carnis were 5.03 µmol/mL and 6.16 µmol/mL; and 6.26 µmol/mL and 6.58 μmol/mL at 10.0 and 25.0% substrate concentration, respectively. The production of cellulase and xylanase was exponential as corn cob concentration increased from 0.5% to 10.0% yielding 586.93 U/mL and 1559.18 U/mL from F. oxysporum, with 590.7 U/mL and 1573.95 U/mL from S. carnis, respectively.

Conclusion: The study shows that the most efficient saccharification of corn cob by F. oxysporum and S. carnis was achieved at 10.0% substrate concentration. This suggests that two separate saccharification processes at this concentration will result in higher yields of enzyme and reducing sugars than a single process involving higher concentration.

Keywords: Saccharification, lignocellulosic wastes, corn cob, reducing sugar, Fusarium oxysporum, Sporothrix carnis.

Olajuyigbe FM, Ogunyewo OA. Comparative evaluation of neglected biomass for efficient and economically viable production of lignocellulolytic enzymes from selected white and soft rot fungi. Curr Biotechnol 2016; 5: 71-80.
Ramasamy S, Balakrishna HS, Selvaraj U, Uppuluri KB. Production and statistical optimization of oxytetracycline from Streptomyces rimosus NCIM 2213 using a new cellulosic substrate, Prosopis juliflora. BioResources 2014; 9: 7209-21.
Jampala P, Murugan P, Ramanujam S, Uppuluri KB. Investigation on the effect of carbon and nitrogen sources for the production of cellulosome by Trichoderma Reesei NCIM 1186 using saturated placket burman design. Biosci Biotechnol Res Asia 2015; 12: 1577-86.
He J, Wu AM, Chen D, et al. Cost-effective lignocellulolytic enzyme production by Trichoderma reesei on a cane molasses medium. Biotechnol Biofuels 2014; 7: 43.
[http://dx.doi.org/10.1186/1754-6834-7-43] [PMID: 24655817]
Sheng T, Zhao L, Gao LF, et al. Lignocellulosic saccharification by a newly isolated bacterium, Ruminiclostridium thermocellum M3 and cellular cellulase activities for high ratio of glucose to cellobiose. Biotechnol Biofuels 2016; 9: 172.
[http://dx.doi.org/10.1186/s13068-016-0585-z] [PMID: 27525041]
Srivastava N, Srivastava M, Mishra PK, et al. Applications of fungal cellulases in biofuel production: Advances and limitations. Renew Sustain Energy Rev 2018; 82: 2379-86.
Kumar SA, Pushpa A. Saccharification by fungi and ethanol production by bacteria using lignocellulosic materials. IRJP 2012; 3: 411-4.
Yoon LW, Ngoh GC, Chua ASM. Simultaneous production of cellulase and reducing sugar through modification of compositional and structural characteristic of sugarcane bagasse. Enzyme Microb Technol 2013; 53(4): 250-6.
[http://dx.doi.org/10.1016/j.enzmictec.2013.05.005] [PMID: 23931690]
Balan V. Current challenges in commercially producing biofuels from lignocellulosic biomass. ISRN Biotechnol 2014. 2014463074
[http://dx.doi.org/10.1155/2014/463074] [PMID: 25937989]
Ogunyewo OA, Olajuyigbe FM. Unravelling the interactions between hydrolytic and oxidative enzymes in degradation of lignocellulosic biomass by Sporothrix carnis under various fermentation conditions. Biochem Res Int 2016. 20161614370
[http://dx.doi.org/10.1155/2016/1614370] [PMID: 26881077]
Olajuyigbe FM, Ogunyewo OA. Enhanced production and physicochemical properties of thermostable crude cellulase from Sporothrix carnis grown on corncob. Biocatal Agric Biotechnol 2016; 7: 110-7.
Olajuyigbe FM, Nlekerem CM, Ogunyewo OA. Production and characterization of highly thermostable β-glucosidase during the biodegradation of methyl cellulose by Fusarium oxysporum. Biochem Res Int 2016; 20163978124
[http://dx.doi.org/10.1155/2016/3978124] [PMID: 26977320]
Olajuyigbe FM. Bioconversion of cellulose and simultaneous production of thermoactive exo- and endoglucanases by Fusarium oxysporum. Cellulose 2017; 24: 4325-36.
Olajuyigbe FM, Fatokun CO. Biochemical characterization of an extremely stable pH-versatile laccase from Sporothrix carnis CPF- 05. Int J Biol Macromol 2017; 94(Pt A): 535-43.
[http://dx.doi.org/10.1016/j.ijbiomac.2016.10.037] [PMID: 27765568]
Van Soest PJ, Robertson JB, Lewis BA. Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J Dairy Sci 1991; 74(10): 3583-97.
[http://dx.doi.org/10.3168/jds.S0022-0302(91)78551-2] [PMID: 1660498]
Wood TM, Bhat KM. Methods in Enzymology: Cellulose and HemicelluloseMethod for measuring cellulase activities. New York: Academic Press 1998; pp. 87-112.
Saha BC. Production, purification and properties of a newly isolated xylanase from Fusarium proliferatum. Process Biochem 2002; 37: 1279-84.
Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 1959; 31: 426-8.
Bajpai P. Pretreatment of lignocellulosic biomass for biofuel production. SpringerBriefs in Green Chemistry for Sustainability 2016.
Sun Y, Cheng J. Hydrolysis of lignocellulosic materials for ethanol production: a review. Bioresour Technol 2002; 83(1): 1-11.
[http://dx.doi.org/10.1016/S0960-8524(01)00212-7] [PMID: 12058826]
Wang L, Yang M, Fan X, Zhu X, Xu T, Yuan Q. An environmentally friendly and efficient method for xylitol bioconversion with high-temperature-steaming corn cob hydrolysate by adapted Candida tropicalis. Process Biochem 2011; 46: 1619-26.
Lu J, Chen W. Product yields and characteristics of corncob waste under various torrefaction atmospheres. Energies 2014; 7: 13-27.
Pointner M, Kuttner P, Obrlik T, Jager A, Kahr H. Composition of corncobs as a substrate for fermentation of biofuels. Agron Res (Tartu) 2014; 12: 391-6.
Chandra RP, Bura R, Mabee WE, Berlin A, Pan X, Saddler JN. Substrate pretreatment: the key to effective enzymatic hydrolysis of lignocellulosics? Adv Biochem Eng Biotechnol 2007; 108: 67-93.
[http://dx.doi.org/10.1007/10_2007_064] [PMID: 17530205]
Carere CR, Sparling R, Cicek N, Levin DB. Third generation biofuels via direct cellulose fermentation. Int J Mol Sci 2008; 9(7): 1342-60.
[http://dx.doi.org/10.3390/ijms9071342] [PMID: 19325807]
Kristensen JB, Felby C, Jørgensen H. Yield-determining factors in high-solids enzymatic hydrolysis of lignocellulose. Biotechnol Biofuels 2009; 2(1): 11.
[http://dx.doi.org/10.1186/1754-6834-2-11] [PMID: 19505292]
Roche CM, Dibble CJ, Stickel JJ. Laboratory-scale method for enzymatic saccharification of lignocellulosic biomass at high-solids loadings. Biotechnol Biofuels 2009; 2(1): 28.
[http://dx.doi.org/10.1186/1754-6834-2-28] [PMID: 19889202]
Qui W. High consistency enzymatic hydrolysis of lignocelluloses. MSc thesis. Vancouver: The University of British Columbia 2010.
Sørensen HR, Pedersen S, Meyer AS. Optimization of reaction conditions for enzymatic viscosity reduction and hydrolysis of wheat arabinoxylan in an industrial ethanol fermentation residue. Biotechnol Prog 2006; 22(2): 505-13.
[http://dx.doi.org/10.1021/bp050396o] [PMID: 16599569]
Cara C, Moya M, Ballesteros I, Negro MJ, Gonzalez A, Ruiz E. Influence of solid loading on enzymatic hydrolysis of steam exploded or liquid hot water pretreated olive tree biomass. Process Biochem 2007; 42: 1003-9.
Rosgaard L, Andric P, Dam-Johansen K, Pedersen S, Meyer AS. Effects of substrate loading on enzymatic hydrolysis and viscosity of pretreated barley straw. Appl Biochem Biotechnol 2007; 143(1): 27-40.
[http://dx.doi.org/10.1007/s12010-007-0028-1] [PMID: 18025594]
Hodge DB, Karim MN, Schell DJ, McMillan JD. Soluble and insoluble solids contributions to high-solids enzymatic hydrolysis of lignocellulose. Bioresour Technol 2008; 99(18): 8940-8.
[http://dx.doi.org/10.1016/j.biortech.2008.05.015] [PMID: 18585030]
Di Risio S, Hu CS, Saville BA, Liao D, Lortie J. Large-scale, high-solids enzymatic hydrolysis of steam-exploded poplar. Biofuels Bioprod Biorefin 2011; 5: 609-20.
Modenbach AA, Nokes SE. The use of high-solids loadings in biomass pretreatment--a review. Biotechnol Bioeng 2012; 109(6): 1430-42.
[http://dx.doi.org/10.1002/bit.24464] [PMID: 22359283]
Kobakhidze A, Asatiani M, Kachlishvili E, Elisashvili V. Induction and catabolite repression of cellulase and xylanase synthesis in the selected white-rot basidiomycetes. Ann Agrar Sci 2016; 14: 169-76.
Fatokun EN, Nwodo UU, Okoh AI. Classical optimization of cellulase and xylanase production by a marine Streptomyces species. Appl Sci (Basel) 2016; 6: 286.
Prakasham RS, Rao ChS, Sarma PN. Green gram husk--an inexpensive substrate for alkaline protease production by Bacillus sp. in solid-state fermentation. Bioresour Technol 2006; 97(13): 1449-54.
[http://dx.doi.org/10.1016/j.biortech.2005.07.015] [PMID: 16140528]
Addela IR, Gujjula R, Surender M, Nyapati S, Rudravaram R, Bhagam R. Production and optimization of xylanase from Penicillium species in solid-state fermentation. Int J Rec Biotech 2015; 3: 15-21.
Abo-State MAM, Hammad AI, Swelin M, Gannam RB. Enhanced production of cellulases by Aspergillus spp. isolated from agriculture wastes by solid-state fermentation. Am Eurasium J Agric Environ Sci 2010; 8: 402-10.
Gautam SP, Budela PS, Pandey AK, Jamaluddin AMK, Sarsaiya S. Optimization of the medium for the production of cellulase by the Trichoderma viride using submerged fermentation. Int J Environ Sci 2010; 4: 656-65.
Jampala P, Tadikamalla S, Preethi M, Ramanujam S, Uppuluri KB. Concurrent production of cellulase and xylanase from Trichoderma reesei NCIM 1186: enhancement of production by desirabilitybased multi-objective method. 3 Biotech 2017; 7(1): 14.
[http://dx.doi.org/10.1007/s13205-017-0607-y] [PMID: 28391478]

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Article Details

Year: 2019
Published on: 20 January, 2020
Page: [109 - 115]
Pages: 7
DOI: 10.2174/2211550108666191008154658

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