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Investigating potential protease activity of psychrotrophic bacteria from a municipal landfill for solid waste management

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A municipal landfill contains different waste materials that support the growth of a diverse community of enzyme-secreting microorganisms that degrade and detoxify the wastes. The present study aimed to explore psychrotrophic protein-degrading bacteria for converting proteinaceous waste into the nutrient-rich end product of agricultural applications. During the study, twenty five morphologically different psychrotrophic proteolytic bacteria were isolated from the landfill soil samples using standard serial dilution and spread plate techniques. The isolates showed variable protease activities at different incubation temperatures (5 °C, 10 °C, 15 °C, and 20 °C). However, isolate PB2 showed significantly (p < 0.05) highest protease activity (1.53 ± 0.8 1U/mL) at 20 °C with corresponding hydrolysis zone of average diameter of 17 ± 2 mm through primary screening. The isolate (PB2) also exhibited the hydrolysis of several protein substrates, including elastin and gelatin. According to protease inhibition studies, the extracellular proteases released by isolating PB2 were primarily serine and metalloproteases. The morphological, biochemical, and molecular characterization (16S rRNA) demonstrated that the isolate had 99% similarity with Pseudomonas fluorescens strain SU003 (NCBI, Accession No. MH266220). The results indicated that psychrotrophic bacteria isolated from the temperate landfill site displayed promising protease activities at lower temperatures and can be utilized for converting voluminous organic wastes generated in Himalayan regions via cold composting to produce nutrient-rich compost of agricultural importance.

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One-way analysis of variance


Duncan’s multiple range test


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Municipal solid waste


National Center for Biotechnology Information




Polymerase chain reaction


Phenylmethylsulfonyl fluoride


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The authors would like to acknowledge the support provided by Researchers Supporting Project Number RSP2024R358, King Saud University, Riyadh, Saudi Arabia. All authors appreciate the Dean of the Faculty of Agriculture Wadura, SKUAST-K, for providing the laboratory facilities required to do microbiological analysis. All authors thank the Director of the Centre of Research for Development at the University of Kashmir for providing laboratory help that allowed the molecular work to go successfully.


The University Grants Commission, New Delhi, financially supported the research through the Junior Research Fellowship under Maulana Azad National Fellowship Scheme under Grant number F1-17.1/2016-2017/MANF-2015-17-JAM-49697.

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BH collected the data, performed the experiments, and prepared the manuscript, ZAB helped in the experimentation and designed the concept, TAS interpreted the data and performed the statistical analysis. KP and RZS performed statistical analysis and wrote, reviewed, and edited the manuscript. All authors read and approved the final manuscript.

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Correspondence to Basharat Hamid or R. Z. Sayyed.

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Hamid, B., Baba, Z.A., Sheikh, T.A. et al. Investigating potential protease activity of psychrotrophic bacteria from a municipal landfill for solid waste management. Biomass Conv. Bioref. (2024).

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