Abstract
Ionic liquids (ILs) have gained immense attention as eco-friendly solvents for pretreatment of lignocellulosics for their potential bioconversion to biofuels, bio-chemicals, and other products. However, for saccharification of ionic liquid (IL)-pretreated biomass, IL-stable cellulases are desired. IL-stable cellulase and xylanase enzyme preparation developed from a previously isolated Aspergillus assiutensis VS34 was used for saccharification of IL-pretreated biomass. Current study reports the purification of IL-stable cellulase (CMCase) from A. assiutensis VS34 based on salt precipitation and ion exchange chromatography. Functionality of the purified cellulase (2.10-fold) was observed by native-PAGE and zymography, and the molecular weight (27 kDa) was assayed by SDS-PAGE. Though optimum temperature and pH of CMCase was 50 ℃ and 6.0, respectively, but the enzyme showed considerable activity and stability over a wide range of temperature (40–80 ℃, 72–99%) and pH (3–11, 60–95%). The activity of enzymes was enhanced by certain metal ions (Ca2+, Cu2+, Mn2+, Mg2+, and Co2+), but decreased considerably by Hg2+. Km and Vmax of CMCase were 6.996 mg/ml and 16.103 μmol/min/mg, respectively. Process-apt properties of A. assiutensis VS34 CMCase reflect its application potential for a variety of processes including biomass conversion.
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Acknowledgements
Dr. Bijender Kumar Bajaj (BKB) gratefully acknowledges the grants received in the form of research projects from ICMR, UGC, DST, CSIR and DBT. BKB gratefully acknowledges various agencies for providing fellowships for overseas ‘Research Stays’ i.e. Indo-US Science and Technology Forum (Ohio State University, USA), Commonwealth Scholarship Commission, UK (Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth, UK), and Institute of Advanced Study, Durham University, UK (COFUND-International Senior Research Fellow at Department of Biosciences, Durham University, UK). Dr. Parushi Nargotra acknowledges Rashtriya Uchchattar Shiksha Abhiyan (RUSA) for providing a Ph.D. research fellowship. Dr. Vishal Sharma acknowledges the DST, Govt. of India for providing Inspire fellowship for doctoral research.
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Financial support in the form of Research Projects to Dr. Bijender Kumar Bajaj (BKB) from funding agencies such as DST, DBT, UGC, CSIR and ICMR is gratefully acknowledged.
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Conceptualization: Vishal Sharma; Methodology: Vishal Sharma; Formal analysis and investigation: Parushi Nargotra; Writing—original draft preparation: Vishal Sharma, Parushi Nargotra; Writing—review and editing: Surbhi Sharma, Ridhika Bangotra, Akhlash P Singh, Nisha Kapoor, Ritu Mahajan; Funding acquisition: Bijender Kumar Bajaj, Resources: Bijender Kumar Bajaj; Supervision: Bijender Kumar Bajaj.
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Sharma, V., Nargotra, P., Sharma, S. et al. Purification and biochemical characterization of an ionic liquid tolerant cellulase from Aspergillus assiutensis VS34 for potential biomass conversion applications. Environmental Sustainability (2024). https://doi.org/10.1007/s42398-024-00311-1
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DOI: https://doi.org/10.1007/s42398-024-00311-1