Biomass Conversion and Biorefinery

, Volume 8, Issue 1, pp 97–111 | Cite as

Binary- and triple-enzyme cocktails and their application mode affect fermentable sugar release from pretreated lignocellulo-starch biomass

  • M.G. Mithra
  • J. Sreekumar
  • G. PadmajaEmail author
Original Article


Lignocellulo-starch biomass (LCSB) comprising roots and vegetable processing wastes has high starch besides cellulose and hemicelluloses and warrants different pretreatment and saccharification approaches. The fermentable sugar yield from steam/dilute sulphuric acid (DSA)-pretreated biomass during saccharification with binary [cellulase + amylolytic enzyme (Stargen)] or triple (cellulase + xylanase + Stargen) enzyme cocktails was compared. The factors such as pH (5.0), temperature (50 °C) and enzyme dosage (16 FPU/g cellulose) for cellulase (Ecozyme RT80) action were optimized using response surface methodology. As pretreated liquor is rich in sugars, whole slurry saccharification was needed for LCSBs and saccharification efficiency (120 h) was significantly higher for steam-pretreated biomass with all application modes. Preferential hydrolysis of starch in steam-pretreated biomass by Stargen followed by cellulolysis was advantageous than the application sequence with cellulase followed by Stargen. Triple-enzyme-based saccharification of steam-pretreated biomass significantly enhanced the overall conversion efficiency (OCE; 85–98%) compared to only 28–49% in the native untreated biomass, while lower OCE was observed in the case of DSA-pretreated and saccharified biomass. Supplementation with both xylanase and Stargen pronouncedly enhanced the OCE for steam-pretreated biomass with only insignificant difference between the exposure periods, indicating the obligatory need for both enzymes for optimal saccharification of LCSBs.


Lignocellulo-starch biomass Pretreatment Saccharification Enzyme cocktails Application mode Fermentable sugars 



The authors acknowledge with gratitude the financial support from the Kerala State Council for Science, Technology & Environment (Grant no. 853/2015/KSCSTE) and the facilities provided by the director, ICAR-CTCRI, for the study. Stargen™ 002 was received by courtesy from M/s Danisco US Inc., USA.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

13399_2017_237_MOESM1_ESM.docx (224 kb)
ESM 1 (DOCX 223 kb)
13399_2017_237_MOESM2_ESM.docx (224 kb)
ESM 2 (DOCX 223 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  1. 1.Division of Crop UtilizationICAR-Central Tuber Crops Research InstituteThiruvananthapuramIndia
  2. 2.Section of Extension and Social SciencesICAR-Central Tuber Crops Research InstituteThiruvananthapuramIndia

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