Abstract
Feather, horn, hoof, and other keratin waste are protein-rich but limited by natural keratinase synthesis, activity, pH, and temperature stability. It is challenging to realize its large-scale application in industries. Bacillus subtilis spores are a safe, efficient, and highly resistant immobilized carrier, which can improve target proteins’ resistance. In this research, KERQ7, the keratinase gene of Bacillus tequilensis strain Q7, was fused to the Bacillus subtilis genes coding for the coat proteins CotG and CotB, respectively, and displayed on the surface of B. subtilis spores. Compared with the free KERQ7, the immobilized KERQ7 showed a greater pH tolerance and heat resistance on the spore surface. The activity of CotG-KERQ7 is 1.25 times that of CotB-KERQ7, and CotG-KERQ7 is more stable. When the flexible linker peptide L3 was used to connect CotG and KERQ7, the activity was increased to 131.2 ± 3.4%, and the residual enzyme activity was still 62.5 ± 2.2% after being kept at 60 ℃ for 4 h. These findings indicate that the flexible linker and CotG were more effective for the spore surface display of keratinase to improve stress resistance and promote its wide application in feed, tanning, washing, and other industries.
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Acknowledgements
This work was supported by laboratories facilities of School of the Life Sciences, Jiangsu University, China.
Funding
This work was supported by the National Key Research and Development Program of China (2021YFC2103004), by the “Jiangsu Funding Program for Excellent Postdoctoral Talent” under Grant number (2023ZB883), and by Key Research and Development Program (Modern Agriculture) of Zhenjiang City, Jiangsu, China (NY2023013).
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ZW conceived and designed this research. ZW and MY conducted the experiments, analyzed the data, and drafted the manuscript. MS, KL, YC, COO, ZF, ZN, and HC revised this manuscript. All the authors read and approved the final manuscript.
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The animal study was reviewed and approved by the Institutional Animal Care and Use Committee of Jiangsu University. (NO: UJS-IACUC-2022030702).
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Wang, Z., Yan, M., Saeed, M. et al. The flexible linker and CotG were more effective for the spore surface display of keratinase KERQ7. World J Microbiol Biotechnol 40, 35 (2024). https://doi.org/10.1007/s11274-023-03854-3
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DOI: https://doi.org/10.1007/s11274-023-03854-3