Improved Production of Multi-component Cellulolytic Enzymes Using Sweet Sorghum Bagasse and Thermophilic Aspergillus terreus RWY Through Statistical Process Optimization

  • Reetika Sharma
  • Gurvinder Singh Kocher
  • Sarvanan Satyanarayana Rao
  • Harinder Singh OberoiEmail author
Original Paper



The study was conducted to improve the productivity of the multi-component cellulolytic enzymes using thermophilic Aspergilus terreus strain and sweet sorghum bagasse as substrate. One of the major objectives was to study the interactions between different operating parameters and appraise the potential of the optimized process for validation studies.


Response surface methodology (RSM) based on central composite design (CCD) was used to optimize the process parameters for cellulase production by thermophilic Aspergillus terreus via a solid-state fermentation (SSF) process. A set of 50 experiments in triplicate with five factors (moisture content, inoculum level, pH, temperature and incubation period), three levels with six axial points (α ± 1.68) and five replications at the central point were conducted in this study with filter paper (FP) cellulase and β-glucosidase as output parameters.


Micrographs and scanning electron microscopy (SEM) of A. terreus RWY revealed a chain of conidia in a columnar arrangement with an average size of conidium being 2.12 µ. Statistical process optimization suggested temperature of 45 °C, pH of 5.8, incubation time of 72 h, inoculum concentration of 10% and initial moisture content of 80% (w/w) as optimum for conducting validation studies. Validation studies showed comparable FP and β-glucosidase activities as predicted by the model equations. In addition to FP and β-glucosidase, A. terreus RWY also produced endoglucanase (EG), β-xylosidase, α-l-arabinofuranosidase, CBHI, xylanase and xylan esterase of 149.54, 26.94, 183.16, 17.52, 1264.47 and 1106.46 U/gds, respectively during the validation process. Response surface optimization also led to a nearly two-fold increase in FP and β-glucosidase activities.


Coupled with the use of thermophilic strains which confer specific benefits during industrial applications, statistical process optimization holds potential for scale-up studies for cellulase production using the optimized parameters, SSB as substrate and thermophilic A. terreus RWY.


Aspergillus terreus β-Glucosidase Filter paper cellulase Response surface methodology Solid state fermentation Sweet sorghum bagasse 



Authors thankfully acknowledge the financial support received under the project (BT/PR8488/PBD/26/68/2006) funded by the Department of Biotechnology (DBT), Government of India for conducting this study. Authors Sharma and Kocher also thankfully acknowledge the support received from the Punjab Agricultural University, Ludhiana, India.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

Research Involving Human Participants and/or Animals

This study did not involve the use of human participants and/or animals.

Supplementary material

12649_2019_670_MOESM1_ESM.docx (610 kb)
Supplementary material 1 (DOCX 610 kb)


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© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of MicrobiologyPunjab Agricultural UniversityLudhianaIndia
  2. 2.ICAR-Central Institute of Post-Harvest Engineering and TechnologyLudhianaIndia
  3. 3.YS Parmar University of Horticulture and ForestryNauniIndia
  4. 4.ICAR-Indian Institute of Millet ResearchHyderabadIndia
  5. 5.ICAR-Indian Institute of Horticultural ResearchBengaluruIndia

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