Abstract
The purpose of the current study was to evaluate the functional activity and storage viability (at 4 °C and 35 °C) of an immobilized as well as lyophilized multienzyme, viz., pectinase, cellulase, and amylase (PCA) that was produced by Bacillus subtilis NG105 under solid state fermentation (SSF) at 35 ℃ for 10 days using mosambi peel as a substrate. After SSF, the culture media was divided into two aliquots. From the first aliquot, the produced ME was extracted, precipitated, and further immobilized on calcium alginate beads (MEICA). In order to immobilize on mosambi peel matrix, the second aliquot was mixed with acetone and subsequently lyophilized (MELMP). Thus, ready MEICA and MELMP extracted 87.5 and 91.5% juice from mango pulp, respectively. In the reusability study, after 5 cycles, MEICA exhibited 23.8%, 24.4%, and 36.5% PCA activity, respectively. The PCA activity of MEICA and MELMP was examined after 60 days of storage at 4 ℃. The result revealed that the PCA for MEICA declined from 100 to 66%, 58.2%, and 64.5%, respectively, while for MELMP, it dropped from 100 to 84.2%, 82.1%, and 69.7%, respectively. Further, after 60 days of storage, the reduction of total protein content (TPC) in free multienzyme (FME), MEICA, and MELMP was 92.2%, 91.5%, and 36.3% observed, respectively. In the localization study, the maximum levels of multienzyme activity were found in cell exudates. This study demonstrated that immobilizing of multienzyme through lyophilization on waste substrates like mosambi peel boosted its stability and shelf-life along with greatly reducing the cost of products.
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Data availability
Sequence information generated was submitted in the NCBI database (Singh et al. 2022a).
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Acknowledgements
The authors would like to thank the Director of ICAR-CISH for providing the necessary facilities for the investigation.
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The research was funded by the “Application of Microorganisms in Agriculture and Allied Sectors” (AMAAS) networking project of the Indian Council of Agricultural Research, New Delhi, India.
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Neelima Garg and Priti Mathur: conceived the research and designed experiments; Balvindra Singh, SupriyaVaish, and Sumit K. Soni: performed the experiments; Balvindra Singh and Sumit K. Soni: analyzed the data and prepared the draft; Neelima Garg, Balvindra Singh, and Sumit K. Soni: editing the draft and prepared the final manuscript.
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Singh, B., Soni, S.K., Vaish, S. et al. Immobilization of microbial multienzyme preparation on calcium alginate beads as well as lyophilization with mosambi peel matrix improved its shelf-life and stability. Folia Microbiol 69, 383–393 (2024). https://doi.org/10.1007/s12223-023-01079-3
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DOI: https://doi.org/10.1007/s12223-023-01079-3