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Identification and Characterization of a Cocoon Degradable Enzyme from the Isolated Strain Bacillus subtilis Bs5C

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  • Enzyme Biotechnology
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Abstract

A fibroin-degrading bacterium was isolated in a medium containing fibroin as a sole nitrogen source, and then identified and characterized. The strain was designated Bacillus subtilis strain Bs5C, and the culture medium containing the extracellular proteolytic enzymes was found to hydrolyze cocoon fiber. A decomposition rate of 35% was achieved with the culture medium in optimized cocoon-degrading condition. In a second round of decomposition performed on the same fibers with additional culture medium, the decomposition rate reached 45.9%. To investigate the proteolytic enzymes degrading cocoon, the enzymes was purified from culture medium using ion exchange column and identified using LC-MS/MS analysis system. As a result, it was uncovered that the cocoon degradation was due to the neutral protease NprE of the strain Bs5C. Moreover, the cocoon fibers treated with semi-purified NprE enzyme solution were clearly split and degraded, as assessed by SEM, and solubilized peptides from fibroin and sericin were detected by LC-MS/MS. In conclusion, this study is the first report that peptides could be produced from cocoon by cultured medium of B. subtilis, and the newly isolated strain Bs5C and the NprE protease from the strain Bs5C could clearly be valuable for the production of silk peptides, which have various pharmacological and industrial applications.

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Acknowledgements

This work was supported by the Korea Research Institute of Bioscience and Biotechnology (KRIBB) Research Initiative Program.

The authors declare that there are no conflicts of interest. Neither ethical approval nor informed consent was required for this study.

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Authors and Affiliations

  1. Industrial Microbiology and Bioprocess Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup, 56212, Korea

    Hyun Deok Oh, Min Seo Cho, Joong-Su Kim, Min-Soo Kim, Chul Ho Kim & Ji Young Kang

  2. Department of Food and Nutrition, Chonnam National University, Gwangju, 61186, Korea

    Hyun Deok Oh

  3. Department of Bioactive Material Science, Chonbuk National University, Jeonju, 54896, Korea

    Min Seo Cho

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  1. Hyun Deok Oh
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Oh, H.D., Cho, M.S., Kim, JS. et al. Identification and Characterization of a Cocoon Degradable Enzyme from the Isolated Strain Bacillus subtilis Bs5C. Biotechnol Bioproc E 25, 442–449 (2020). https://doi.org/10.1007/s12257-019-0399-5

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