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Secretome Analysis and Bioprospecting of Lignocellulolytic Fungal Consortium for Valorization of Waste Cottonseed Cake by Hydrolase Production and Simultaneous Gossypol Degradation

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Abstract

Purpose

The utilisation of agro-wastes as feedstocks for biorefinery development is a promising strategy to combat the looming energy challenges with minimal environmental impact. Deoiled cottonseed cake is an abundant lignocellulosic residual agro-waste with restricted usage due to presence of toxic gossypol. The utilisation of this waste biomass for production of hydrolases (cellulase and xylanase) and simultaneous degradation of toxic gossypol was attempted by fungal mediated bioprocess development. The secretome profiling was carried out to gain insights into the diversity of proteins expressed by the fungal consortium under solid-state fermentation (SSF) conditions.

Methods

The three potential lignocellulolytic fungal strains, Aspergillus niger, Trichoderma reesei, Phanerochaete chrysosporium and their consortium were exploited for SSF of cottonseed cake. The secretome profiling was carried out by LC-ESI MS/MS based proteomic approach. The efficiency of hydrolases produced from seed cake was verified by saccharification of wheat straw.

Results

The consortium produced highest titre of cellulase (FPase: 21.62 IU; CMCase: 155.41 IU; β-glucosidase: 29.73 IU) and xylanase (2008.05 IU) per gram cake with 90% reduction in gossypol content. The secretome analysis revealed versatile mixture of 81 proteins comprising of hemicellulases (36%), cellulases (17%), amylases (7%), esterases (6%), proteases (5%), hypothetical proteins (20%) and other proteins (9%). The Glycosyl hydrolase (GH) proteins constituted 73% of the total secretome. The high yield of reducing sugars (402.38 mg/g) was obtained from saccharification of wheat straw.

Conclusions

The present study highlights the sustainable valorization of an agro-waste by microbiological transformation for production of value-added products in biorefinery.

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Acknowledgements

Authors gratefully acknowledge the financial grant provided by Department of Science and Technology (DST/INT/TUNISIA/P07/2017) and the Ministry of Food Processing Industries (Govt. of India) for carrying out this study. Jasneet Grewal is grateful to Indian Institute of Technology Delhi for her Senior Research Fellowship and providing the infrastructural facilities.

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  1. Enzyme and Microbial Biochemistry Laboratory, Department of Chemistry, Indian Institute of Technology Delhi, Hauz Khas, New Delhi, 110016, India

    Jasneet Grewal, Rameshwar Tiwari & S. K. Khare

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  1. Jasneet Grewal
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12649_2019_620_MOESM1_ESM.docx

Supplementary Fig.S1 SEM image of (a) unfermented cottonseed cake (control) (b) A. niger (c) T. reesei (d) P. chrysosporium (e) consortium growing on cottonseed cake at ×2500 magnification (DOCX 1104 KB)

12649_2019_620_MOESM2_ESM.docx

Supplementary Fig.S2 HPLC chromatogram of extracted gossypol-diamino-propanol (stable schiff’s base derivative) from cottonseed cake (a) unfermented (control) (b) fermented by P. chyrosporium (c) fermented by T. reesei (d) fermented by A. niger (e) fermented by consortium (DOCX 642 KB)

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Grewal, J., Tiwari, R. & Khare, S.K. Secretome Analysis and Bioprospecting of Lignocellulolytic Fungal Consortium for Valorization of Waste Cottonseed Cake by Hydrolase Production and Simultaneous Gossypol Degradation. Waste Biomass Valor 11, 2533–2548 (2020). https://doi.org/10.1007/s12649-019-00620-1

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