Abstract
Experimental investigations were carried out to develop economic production process of cellulase using coconut mesocarp as an inexpensive lignocellulosic inducer while replacing commercial cellulose. Cellulase production was initially investigated from commercial cellulose in different submerged conditions using Trichoderma reesei (MTCC 164). Maximum enzyme production was achieved 6.3 g/l with activity level 37 FPU/ml in the condition where cellulose to water content ratio was maintained at 5:35 (W/V). To achieve similar maximum production of cellulase from coconut mesocarp, response surface methodology was implemented to optimize most influencing parameters. Most influencing nutritional parameters such as coconut mesocarp, glucose and peptone were optimized in the concentration ranges of 35 g/l, 35 g/l and 25 g/l, respectively. Selecting optimized parameter values, fermentations were conducted inside the fermenter with 2 L operating volume to ensure high concentration and activity profiles of enzyme. Enzyme concentration was achieved 7.20 g/l after 96 h of batch fermentation with specific activity levels of 42 FPU/ ml and CMCase 75 U/ml. Enzyme concentration was further improved to 9.58 g/l with activity levels of 54 FPU/ml and CMCase 93 U/ml by adopting sequential feeding of coconut mesocarp in fed-batch fermentation mode. The presence of pure cellulase in the sample was confirmed by FTIR analysis.
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References
Ahamed A, Vermette P (2010) Effect of mechanical agitation on the production of cellulases by Trichoderma reesei RUT-C30 in a draft-tube airlift bioreactor. Biochem Eng J 49:379–387. https://doi.org/10.1016/J.BEJ.2010.01.014
Avelino F, Alves G, Ruiz HA et al (2015) Bioethanol production from coconuts and cactus pretreated by autohydrolysis. Ind Crops Prod 77:1–12. https://doi.org/10.1016/j.indcrop.2015.06.041
Ballesteros I, Negro MJ, Oliva JM et al (2006) Ethanol production from steam-explosion pretreated wheat straw. Appl Biochem Biotechnol 129–132:496–508
Belghith H, Ellouz-Chaabouni S, Gargouri A (2001) Biostoning of denims by Penicillium occitanis (Pol6) cellulases. J Biotechnol 89:257–262
Bendig C, Weuster-Botz D (2013) Reaction engineering analysis of cellulase production with Trichoderma reesei RUT-C30 with intermittent substrate supply. Bioprocess Biosyst Eng 36:893–900. https://doi.org/10.1007/s00449-012-0822-1
Bohara RA, Thorat ND, Pawar SH (2016) Immobilization of cellulase on functionalized cobalt ferrite nanoparticles. Korean J Chem Eng 33:216–222. https://doi.org/10.1007/s11814-015-0120-0
Cabral MMS, Abud de AKS, Silva de CEF et al (2016) Bioethanol production from coconut husk fiber. Ciência Rural 46:1872–1877. https://doi.org/10.1590/0103-8478cr20151331
Cen P, Xia L (1999) Production of cellulase by solid-state fermentation. Springer, Berlin, pp 69–92
Cunha FM, Esperança MN, Zangirolami TC et al (2012) Sequential solid-state and submerged cultivation of Aspergillus niger on sugarcane bagasse for the production of cellulase. Bioresour Technol 112:270–274. https://doi.org/10.1016/J.BIORTECH.2012.02.082
De Cassia Pereira J, Marques NP, Rodrigues A et al (2015) Thermophilic fungi as new sources for production of cellulases and xylanases with potential use in sugarcane bagasse saccharification. J Appl Microbiol. https://doi.org/10.1111/jam.12757
Dey P, Rangarajan V (2017) Improved fed-batch production of high-purity PHB (poly-3 hydroxy butyrate) by Cupriavidus necator (MTCC 1472) from sucrose-based cheap substrates under response surface-optimized conditions. 3 Biotech. https://doi.org/10.1007/s13205-017-0948-6
Domingues FC, Queiroz JA, Cabral JMS, Fonseca LP (2001) Production of cellulases in batch culture using a mutant strain of Trichoderma reesei growing on soluble carbon source. Biotechnol Lett 23:771–775. https://doi.org/10.1023/A:1010329731010
Ebrahimi M, Caparanga AR, Ordono EE, Villaflores OB (2017) Evaluation of organosolv pretreatment on the enzymatic digestibility of coconut coir fibers and bioethanol production via simultaneous saccharification and fermentation. Renew Energy 109:41–48. https://doi.org/10.1016/j.renene.2017.03.011
Esterbauer H, Steiner W, Labudova I et al (1991) Production of Trichoderma cellulase in laboratory and pilot scale. Bioresour Technol 36:51–65. https://doi.org/10.1016/0960-8524(91)90099-6
FAOSTAT Data (2016) The top 5 coconut producing countries. In: FAOSTAT data, 2016 (last accessed by Top 5 Anything January 2016). https://top5ofanything.com/list/1cb15034/Coconut-Producing-Countries (Accessed 1 Aug 2017). Accessed 23 May 2018
Ghose TK (1987) Measurement of cellulase activities. Pure Appl Chem 59:257–268. https://doi.org/10.1351/pac198759020257
Gonçalves FA, Ruiz HA, Nogueira da CC, et al (2014) Comparison of delignified coconuts waste and cactus for fuel-ethanol production by the simultaneous and semi-simultaneous saccharification and fermentation strategies. Fuel 131:66–76. https://doi.org/10.1016/j.fuel.2014.04.021
Gonçalves FA, Ruiz HA, Silvino dos Santos E et al (2016) Bioethanol production by Saccharomyces cerevisiae, Pichia stipitis and Zymomonas mobilis from delignified coconut fibre mature and lignin extraction according to biorefinery concept. Renew Energy 94:353–365. https://doi.org/10.1016/j.renene.2016.03.045
Hansen GH, Lübeck M, Frisvad JC et al (2015) Production of cellulolytic enzymes from ascomycetes: comparison of solid state and submerged fermentation. Process Biochem 50:1327–1341. https://doi.org/10.1016/J.PROCBIO.2015.05.017
Hasunuma T, Okazaki F, Okai N et al (2013) A review of enzymes and microbes for lignocellulosic biorefinery and the possibility of their application to consolidated bioprocessing technology. Bioresour Technol 135:513–522. https://doi.org/10.1016/j.biortech.2012.10.047
Hendy NA, Wilke CR, Blanch HW (1984) Enhanced cellulase production in fed-batch culture of Trichoderma reesei C30. Enzyme Microb Technol 6:73–77. https://doi.org/10.1016/0141-0229(84)90038-3
Howard RL, Abotsi E, Jansen van REL, Howard S (2003) Lignocellulose biotechnology: issues of bioconversion and enzyme production. Afr J Biotechnol 2:602–619. https://doi.org/10.5897/AJB2003.000-1115
Ito K, Yoshida K, Ishikawa T, Kobayashi S (1990) Volatile compounds produced by the fungus Aspergillus oryzae in rice Koji and their changes during cultivation. J Ferment Bioeng 70:169–172. https://doi.org/10.1016/0922-338X(90)90178-Y
Juhász T, Szengyel Z, Réczey K et al (2005) Characterization of cellulases and hemicellulases produced by Trichoderma reesei on various carbon sources. Process Biochem 40:3519–3525. https://doi.org/10.1016/J.PROCBIO.2005.03.057
Kataria R, Ghosh S (2011) Saccharification of Kans grass using enzyme mixture from Trichoderma reesei for bioethanol production. Bioresour Technol 102:9970–9975. https://doi.org/10.1016/j.biortech.2011.08.023
Kataria R, Ruhal R, Babu R, Ghosh S (2013) Saccharification of alkali treated biomass of Kans grass contributes higher sugar in contrast to acid treated biomass. Chem Eng J 230:36–47. https://doi.org/10.1016/j.cej.2013.06.045
Kim I, Han J-I (2012) Optimization of alkaline pretreatment conditions for enhancing glucose yield of rice straw by response surface methodology. Biomass Bioenerg 46:210–217. https://doi.org/10.1016/J.BIOMBIOE.2012.08.024
Kovács K, Szakacs G, Zacchi G (2009) Comparative enzymatic hydrolysis of pretreated spruce by supernatants, whole fermentation broths and washed mycelia of Trichoderma reesei and Trichoderma atroviride. Bioresour Technol 100:1350–1357. https://doi.org/10.1016/j.biortech.2008.08.006
Li Y, Liu C, Bai F, Zhao X (2016) Overproduction of cellulase by Trichoderma reesei RUT C30 through batch-feeding of synthesized low-cost sugar mixture. Bioresour Technol 216:503–510. https://doi.org/10.1016/j.biortech.2016.05.108
Loaces I, Schein S, Noya F (2017) Ethanol production by Escherichia coli from Arundo donax biomass under SSF, SHF or CBP process configurations and in situ production of a multifunctional glucanase and xylanase. Bioresour Technol 224:307–313. https://doi.org/10.1016/j.biortech.2016.10.075
Lowry OH, Rosebrough NJ, Randall FRJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Lv S, Yu Q, Zhuang X et al (2013) The influence of hemicellulose and lignin removal on the enzymatic digestibility from sugarcane bagasse. Bio Energy Res 6:1128–1134. https://doi.org/10.1007/s12155-013-9297-4
Ma L, Li C, Yang Z et al (2013) Kinetic studies on batch cultivation of Trichoderma reesei and application to enhance cellulase production by fed-batch fermentation. J Biotechnol 166:192–197. https://doi.org/10.1016/J.JBIOTEC.2013.04.023
Martinez D, Berka RM, Henrissat B et al (2008) Genome sequencing and analysis of the biomass-degrading fungus Trichoderma reesei (syn. Hypocrea jecorina). Nat Biotechnol 26:553–560. https://doi.org/10.1038/nbt1403
Menon V, Rao M (2012) Trends in bioconversion of lignocellulose: biofuels, platform chemicals and biorefinery concept. Prog Energy Combust Sci 38:522–550. https://doi.org/10.1016/j.pecs.2012.02.002
Miftahul Jannah A, Asip F (2015) Bioethanol production from coconut fiber using alkaline pretreatment and acid hydrolysis method. Int J Adv Sci Eng Inf Technol 5:320. https://doi.org/10.18517/ijaseit.5.5.570
Miller GL, Miller G (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar no title. Anal Chem 31:426–428. https://doi.org/10.1021/ac60147a030
Mishra A, Sardar M (2015) Cellulase assisted synthesis of nano-silver and gold: application as immobilization matrix for biocatalysis. Int J Biol Macromol 77:105–113. https://doi.org/10.1016/j.ijbiomac.2015.03.014
Omojasola PF, Jilani O et al (2008) Cellulase production by some fungi cultured on pineapple waste. Nat Sci 6:64–79
Rana V, Eckard AD, Teller P, Ahring BK (2014) On-site enzymes produced from Trichoderma reesei RUT-C30 and Aspergillus saccharolyticus for hydrolysis of wet exploded corn stover and loblolly pine. Bioresour Technol 154:282–289. https://doi.org/10.1016/J.BIORTECH.2013.12.059
Rawat R, Kumbhar BK, Tewari L (2013) Optimization of alkali pretreatment for bioconversion of poplar (Populus deltoides) biomass into fermentable sugars using response surface methodology. Ind Crops Prod 44:220–226. https://doi.org/10.1016/J.INDCROP.2012.10.029
Sajith S, Priji P, Sreedevi S, Benjamin S (2016) An overview on fungal cellulases with an industrial perspective. J Nutr Food Sci 6:1–13. https://doi.org/10.4172/2155-9600.1000461
Sateesh L, Rodhe AV, Naseeruddin S et al (2012) Simultaneous cellulase production, saccharification and detoxification using dilute acid hydrolysate of S. spontaneum with Trichoderma reesei NCIM 992 and Aspergillus niger. Indian J Microbiol 52:258–262. https://doi.org/10.1007/s12088-011-0184-4
Singhania RR, Sukumaran RK, Patel AK et al (2010) Advancement and comparative profiles in the production technologies using solid-state and submerged fermentation for microbial cellulases. Enzyme Microb Technol 46:541–549. https://doi.org/10.1016/J.ENZMICTEC.2010.03.010
Sluiter A, Hames B, Ruiz R, Scarlata C, Sluiter J, Templeton D, Crocker D et al (2008) Determination of structural carbohydrates and lignin in biomass. Lab Anal Proced 1617:1–16
Soares J, Demeke MM, Foulquié-Moreno MR et al (2016) Green coconut mesocarp pretreated by an alkaline process as raw material for bioethanol production. Bioresour Technol 216:744–753. https://doi.org/10.1016/j.biortech.2016.05.105
Sun S, Li S, Avera BN et al (2017) Soil bacterial and fungal communities show distinct recovery patterns during forest ecosystem restoration. Appl Environ Microbiol 83:e00966–e00917. https://doi.org/10.1128/AEM.00966-17
Ximenes EA, Dien BS, Ladisch MR et al (2007) Enzyme production by industrially relevant fungi cultured on coproduct from corn dry grind ethanol plants. Appl Biochem Biotechnol 137–140:171–183. https://doi.org/10.1007/s12010-007-9049-z
Yang S-T (2007) Bioprocessing for value-added products from renewable resources: new technologies and applications. Elsevier, New York
Zdarta J, Jędrzak A, Klapiszewski Ł, Jesionowski T (2017) Immobilization of cellulase on a functional inorganic–organic hybrid support: stability and kinetic study. Catalysts 7:374. https://doi.org/10.3390/catal7120374
Zhuang X, Wang W, Yu Q et al (2016) Liquid hot water pretreatment of lignocellulosic biomass for bioethanol production accompanying with high valuable products. Bioresour Technol 199:68–75. https://doi.org/10.1016/j.biortech.2015.08.051
Acknowledgements
The research work was financially supported by Short Term research Grant for the faculty members (Sanctioned Letter: KU/AR/KSTG/32/2017, Sr no 9), Karunya Institute of Technology and Sciences (Deemed to be university). Authors are also thankful to each other for their individual support and contribution towards the completion of the research work and writing the paper.
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Dey, P., Singh, J., Scaria, J. et al. Improved production of cellulase by Trichoderma reesei (MTCC 164) from coconut mesocarp-based lignocellulosic wastes under response surface-optimized condition. 3 Biotech 8, 402 (2018). https://doi.org/10.1007/s13205-018-1421-x
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DOI: https://doi.org/10.1007/s13205-018-1421-x