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Screening and Optimization of Newly Isolated Thermotolerant Lysinibacillus fusiformis Strain SK for Protease and Antifungal Activity

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Abstract

The current study was designed to isolate, identify and characterize a Bacillus sp. capable of producing protease and exhibiting antifungal activity. A highly potent bacterium capable of producing protease abundantly was isolated from the soil collected from the waste pit near Microbiology Laboratory of Birendra Multiple Campus, Bharatpur and later on identified as Lysinibacillus fusiformis strain SK on the basis of morphological, physiological, biochemical and 16S rDNA gene sequencing techniques. The strain SK showed 98.36% similarity with L. fusiformis strain NBRC 15717. Using R-programming statistical analysis tool, the optimum incubation time for the highest average protease production (APP) (47.2 U/mL) was found to be 22 h at 50 °C and both incubation time and temperature showed a significant impact on the production of protease (P < 0.01). The maximum average relative protease activity (ARPA) was observed at pH 7.8 and 48 °C, whereas the least ARPA was observed in the presence of 80 g/L NaCl and 10 g/L HgCl2 (P  < 0.01). The newly isolated bacterial strain also exhibited strong antifungal activity against aflatoxigenic Aspergillus flavus and Aspergillus parasiticus suggesting that it can be a potential candidate for protease production and activity over a wider range of temperature and pH as well as for synthesizing effective antifungal compounds.

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Authors and Affiliations

  1. Department of Microbiology, Birendra Multiple Campus, Tribhuvan University, Chitwan, Bharatpur, 44200, Nepal

    Sujan Khadka, Sanjib Adhikari, Sanjeep Sapkota & Ramesh Sharma Regmi

  2. State Key Laboratory of Environmental Aquatic Chemistry, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, 100085, China

    Sujan Khadka

  3. State Key Laboratory of Alpine Ecology and Biodiversity, Institute of Tibetan Plateau Research, Chinese Academy of Sciences, Beijing, 100101, China

    Alina Thapa & Namita Paudel Adhikari

  4. Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, China

    Raju Panday & Manjila Adhikari

  5. State Key Laboratory of Respiratory Disease, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530, China

    Sanjeep Sapkota

  6. Key Laboratory of Mountain Surface Process and Ecological Regulation, Institute of Mountain Hazards and Environment, Chinese Academy of Sciences, Chengdu, 610041, China

    Ram Proshad

  7. University of Chinese Academy of Sciences, Beijing, 100049, China

    Sujan Khadka, Alina Thapa, Sanjeep Sapkota, Namita Paudel Adhikari & Ram Proshad

  8. Department of Natural Products Research, Dr. Koirala Research Institute for Biotechnology and Biodiversity, Kathmandu, 44613, Nepal

    Niranjan Koirala

  9. Faculty of Science and Technology, University of Macau, Macau SAR, 999078, China

    Niranjan Koirala

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  1. Sujan Khadka
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Khadka, S., Adhikari, S., Thapa, A. et al. Screening and Optimization of Newly Isolated Thermotolerant Lysinibacillus fusiformis Strain SK for Protease and Antifungal Activity. Curr Microbiol 77, 1558–1568 (2020). https://doi.org/10.1007/s00284-020-01976-7

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