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Biochemical Characterization and Application of Thermostable-Alkaline Keratinase From Bacillus halodurans SW-X to Valorize Chicken Feather Wastes

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Abstract

The production, purification, and characterization of thermostable-alkaline keratinase from Bacillus halodurans SW-X (keratinase_SW-X), and its applicability towards the production of white chicken feather hydrolysate and bioplastic starch/keratin production were investigated. A maximal keratinase activity of 16.05 ± 0.25 U/mL was achieved by cultivation of B. halodurans SW-X in 5 L stirred tank bioreactor at 47 °C, 200 rpm, and 2 vvm for 48 h. Keratinase_SW-X was purified as a monomeric protein with the estimated MW of 25 kDa, 14.4-fold, and 20% recovery yield. The protein was classified as thermostable-alkaline keratinase due to its optimal pH (pH 10) and temperature (70 °C), pH stability (pH 9.0–11), thermostability (> 3 h at 50–60 °C), and it showed the highest specificity towards chicken feather keratin with Km of 0.45 mg/mL, and Vmax of 3.51 μmol/min/mg. Crude keratinase_SW-X biodegraded white chicken feathers into two types of low (10–20 kDa) and high (≥ 30 kDa) chicken feather (keratin) hydrolysates, with total recovery yields of 30%. The bioplastic starch/keratin was synthesized using cassava starch (70%, w/w), glycerol (30%, w/w), and keratin hydrolysate (0–0.30%, w/w). The optimal concentration of keratin hydrolysate was 0.10% (w/w), resulting in improved strength and elongation at break, with a good appearance of the bioplastic starch/keratin. Our study suggests that B. halodurans SW-X is a potential strain for thermostable-alkaline keratinase production. Moreover, from environmental perspectives, keratinases_SW-X is a promising candidate enzyme that valorizes low cost chicken feather waste to a high value added product, indicating a novel approach for feather waste treatment and utilization.

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Data availability all data generated or analyzed during this study are included in this published article and its supplementary information files.

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Acknowledgements

The authors gratefully acknowledge the financial support from the National Research Council of Thailand (NRCT) through the Royal Golden Jubilee Ph.D. Programme, Thailand (Grant No. PHD/0144/2560) to Tanyawat Kaewsalud and Thanongsak Chaiyaso. This research was also supported by CMU Mid-Career Research Fellowship program; the Biotechnology Program, Graduate School of Chiang Mai University; the Cluster of Agro Bio-Circular-Green Industry (Agro BCG); Faculty of Agro-Industry; Chiang Mai University, Thailand; and the Japan Student Services Organization (JASSO) student exchange support program (Scholarship for Short-Term Study in Japan) FY2019.

Funding

This work was financially supported by the National Research Council of Thailand (NRCT) in the framework of the Royal Golden Jubilee Ph.D. Programme (Grant No. PHD/0144/2560) to Tanyawat Kaewsalud and Thanongsak Chaiyaso, and supported by the CMU Mid-Career Research Fellowship program (Grant Year 2020); the Japan Student Services Organization (JASSO) student exchange support program (Scholarship for Short-Term Study in Japan) FY2019.

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Authors and Affiliations

  1. Interdisciplinary Program in Biotechnology, Graduate School, Chiang Mai University, Chiang Mai, 50200, Thailand

    Tanyawat Kaewsalud & Kamon Yakul

  2. Division of Packaging Technology, Faculty of Agro-Industry, Chiang Mai University, Chiang Mai, 50100, Thailand

    Kittisak Jantanasakulwong

  3. Department of Animal and Aquatic Sciences, Faculty of Agriculture, Chiang Mai University, Chiang Mai, 50200, Thailand

    Wanaporn Tapingkae

  4. Graduate School of Agriculture, Yamagata University, 1-23 Wakaba-machi, Tsuruoka, Yamagata, 997-8555, Japan

    Masanori Watanabe

  5. Division of Biotechnology, Faculty of Agro-Industry, Chiang Mai University, Chiang Mai, 50100, Thailand

    Thanongsak Chaiyaso

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  1. Tanyawat Kaewsalud
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Kaewsalud, T., Yakul, K., Jantanasakulwong, K. et al. Biochemical Characterization and Application of Thermostable-Alkaline Keratinase From Bacillus halodurans SW-X to Valorize Chicken Feather Wastes. Waste Biomass Valor 12, 3951–3964 (2021). https://doi.org/10.1007/s12649-020-01287-9

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