Energy, Ecology and Environment

, Volume 3, Issue 2, pp 126–135 | Cite as

Combinatorial application of ammonium carbonate and sulphuric acid pretreatment to achieve enhanced sugar yield from pine needle biomass for potential biofuel–ethanol production

  • Surbhi Vaid
  • Neha Bhat
  • Parushi Nargotra
  • Bijender Kumar Bajaj
Original Article


Lignocellulosic biomass (LB) despite its huge potential as a renewable bioenergy resource faces bottlenecks due to its recalcitrance and lack of appropriate pretreatment approaches. The current study evaluates the combinatorial application of alkali and acid pretreatment of pine needle biomass (PNB), for achieving high sugar release upon enzymatic saccharification. Pine needle accumulation poses a big threat to the forest soil fertility and overall ecosystem and environment. However, pine needle waste can be valorized after appropriate pretreatment and enzymatic saccharification for production of renewable energy, i.e. biofuel–ethanol. In combinatorial pretreatment strategy, first PNB was subjected to ammonium carbonate pretreatment, and parameters like ammonium carbonate concentration, incubation time and pretreatment temperature were optimized using design of experiment (DoE) approach. The relative influence of parameters on efficacy of pretreatment was established individually and in interactive terms. Based on DoE, sugar yield of 7.56 mg/g of PNB was obtained. Furthermore, DoE-based pretreated PNB was subjected to sulphuric acid pretreatment, followed by enzymatic saccharification. The sugar released during various steps was pooled (8.19 g/100 g), concentrated and subjected to ethanol fermentation with dual yeast cultures using Saccharomyces cerevisiae and Pichia stipitis. An ethanol yield of 8.8%, v/v (6.94% w/v), was obtained. This represents the process efficiency of 19.34% for bioethanol production from PNB.


Pine needle biomass Ammonium carbonate–sulphuric acid pretreatment Enzymatic saccharification Fermentation Bioethanol 



Dr. Bijender Kumar (Bajaj) gratefully acknowledges the Institute of Advanced Study, Durham University, UK, for providing COFUND International Senior Research Fellowship for ‘Research Stay’ at Department of Biosciences, Durham University, Durham, UK; Department of Science and Technology (Govt. of India) is acknowledged for financial support (Research Project Ref. SR/SO/BB-66/2007), and Commonwealth Scholarship Commission, UK, for providing Commonwealth Fellowship (INCF-2013-45) for ‘Research Stay’ at Institute of Biological, Environmental and Rural Sciences (IBERS), Aberystwyth University, Aberystwyth, UK. Authors thank the Director, School of Biotechnology, University of Jammu, Jammu, for necessary laboratory facilities.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Joint Center on Global Change and Earth System Science of the University of Maryland and Beijing Normal University and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Surbhi Vaid
    • 1
  • Neha Bhat
    • 1
  • Parushi Nargotra
    • 1
  • Bijender Kumar Bajaj
    • 1
    • 2
  1. 1.School of BiotechnologyUniversity of JammuJammuIndia
  2. 2.Department of BiosciencesDurham UniversityDurhamUK

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