Improved production of cellulase by Trichoderma reesei (MTCC 164) from coconut mesocarp-based lignocellulosic wastes under response surface-optimized condition
- 108 Downloads
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.
KeywordsCellulase Coconut mesocarp Cellulose Response surface methodology Fed-batch cultivation
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.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interests.
- Cen P, Xia L (1999) Production of cellulase by solid-state fermentation. Springer, Berlin, pp 69–92Google Scholar
- 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 CrossRefPubMedPubMedCentralGoogle Scholar
- 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 CrossRefGoogle Scholar
- 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
- 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 CrossRefGoogle Scholar
- 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 CrossRefGoogle Scholar
- 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 CrossRefPubMedGoogle Scholar
- 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 CrossRefPubMedGoogle Scholar
- Lowry OH, Rosebrough NJ, Randall FRJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
- Omojasola PF, Jilani O et al (2008) Cellulase production by some fungi cultured on pineapple waste. Nat Sci 6:64–79Google Scholar
- 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 CrossRefPubMedGoogle Scholar
- 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 CrossRefPubMedGoogle Scholar
- 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 1617:1–16Google Scholar
- Yang S-T (2007) Bioprocessing for value-added products from renewable resources: new technologies and applications. Elsevier, New YorkGoogle Scholar