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Harnessing the Keratinolytic Activity of Bacillus licheniformis Through Random Mutagenesis Using Ultraviolet and Laser Irradiations

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Abstract

Keratinase is one of the important proteases, which is widely used for converting keratin of the keratinaceous materials into various value-added products. In this study, a popular keratinase producer, Bacillus licheniformis PWD-1, was exposed to ultraviolet (UV) and He–Ne laser irradiations to develop high keratinase-producing mutants. Laser irradiation showed a higher lethality of cells (94%) than UV treatment (92%), whereas laser treatment required a longer time (75 min) than UV treatment (20 min). A total of 58 mutants were selected from 176 isolates to study protein and keratinase production capability of the mutants. The highest keratin-to-casein (K:C) ratio (1.43) was exhibited by LU11 mutant, which was obtained from the combined laser and UV irradiations. The purified keratinase (65 kDa) of LU11 showed 40% yield 1.7-fold purity, while the respective value for wild enzyme was 29% and 1.3-fold. Both enzymes showed optimal activity at 55 ℃ and pH 8, with a Z value of 15.78 ℃ for LU11 and 19.72 ℃ for wild strain. The Vmax and specific constant (Kcat/Km) of the mutant enzyme were 357.17 U/ml and 33.11 min−1 mM−1, respectively, which were significantly higher than the respective values of wild enzyme (102.04 U/ml and 28.36 min−1 mM−1).

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Data Availability

The dataset used and/or analyzed during this study are available from the corresponding author on reasonable request.

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Funding

This work was supported by the National Natural Science Foundation of China (Grant No.: 31601516).

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Authors and Affiliations

  1. School of Food and Biological Engineering, Jiangsu University, 301 Xuefu Road, Zhenjiang, 212013, Jiangsu, China

    Jamila Akter Tuly, Haile Ma, Hossain M. Zabed, Yating Dong, Quaisie Janet, Lu Feng, Ting Li & Gu Chen

  2. Key Laboratory for Physical Processing of Agricultural Products, Jiangsu University, Zhenjiang, China

    Haile Ma

  3. Faculty of Applied Sciences and Technology, Sunyani Technical University, P.O. Box 206, Sunyani, Ghana

    Moses Kwaku Golly

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  1. Jamila Akter Tuly
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  2. Haile Ma
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  3. Hossain M. Zabed
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Contributions

JAT: conceptualization, investigation, experimentation, and original draft preparation; HM: conceptualization, manuscript reviewing, and supervision; HMZ: conceptualization and editing the manuscript; YD: data analysis; QJ: manuscript reviewing; MKG: manuscript reviewing; LF: validation and data analysis; TL: data analysis; GC: data analysis and resource management.

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Correspondence to Haile Ma.

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Tuly, J.A., Ma, H., Zabed, H.M. et al. Harnessing the Keratinolytic Activity of Bacillus licheniformis Through Random Mutagenesis Using Ultraviolet and Laser Irradiations. Appl Biochem Biotechnol 194, 1546–1565 (2022). https://doi.org/10.1007/s12010-021-03697-4

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