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Modular Engineering to Enhance Keratinase Production for Biotransformation of Discarded Feathers

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Abstract

Biotransformation of wasted feathers via feather-degrading enzyme has gained immense popularity, low conversion efficiency hinders its scale application, and the main purpose of this study is to improve feather-degrading enzyme production in Bacillus licheniformis. Firstly, keratinase from Bacillus amyloliquefaciens K11 was attained with the best performance for feather hydrolysis, via screening several extracellular proteases from Bacillus; also, feather powder was proven as the most suitable substrate for determination of feather-degrading enzyme activity. Then, expression elements, including signal peptides and promoters, were optimized, and the combination of signal peptide SPSacC with promoter Pdual3 owned the best performance, keratinase activity aggrandized by 6.21-fold. According to amino acid compositions of keratinase and feeding assays, Ala, Val, and Ser were proven as critical precursors, and strengthening these precursors’ supplies via metabolic pathway optimization resulted in a 33.59% increase in the keratinase activity. Furthermore, keratinase activity reached 2210.66 U/mL, up to 56.74-fold from the original activity under the optimized fermentation condition in 3-L fermentor. Finally, the biotransformation process of discarded feathers by the fermented keratinase was optimized, and our results indicated that 90.94% of discarded feathers (16%, w/v) were decomposed in 12 h. Our results suggested that strengthening precursor amino acids’ supplies was an efficient strategy for enhanced production of keratinase, and this research provided an efficient strain as well as the biotransformation process for discarded feather re-utilization.

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All data generated and analyzed during this study are included in this article.

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Funding

This work was supported by the National Key Research and Development Program of China (2021YFC2100200), the Natural Science Foundation of Fujian Province (2021J01607), the Open Project Funding of Key Laboratory of Green Chemical Technology of Fujian Province University (WYKF-GCT2020-3), and the Open Project Funding of the State Key Laboratory of Biocatalysis and Enzyme Engineering (SKLBEE2018002).

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Authors and Affiliations

  1. State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, 368 Youyi Avenue, Wuchang District, Wuhan, 430062, Hubei, People’s Republic of China

    Yongqing Liao, Min Xiong, Zhaoqi Miao, Ali Raza Ishaq, Yangyang Zhan, Dongbo Cai, Zhifan Yang & Shouwen Chen

  2. Key Laboratory of Green Chemical Technology of Fujian Province University, College of Ecological and Resource Engineering, Wuyi University, Wuyishan, 354300, People’s Republic of China

    Min Zhang, Bichan Li & Shouwen Chen

  3. School of Chemistry and Chemical Engineering, Wuhan University of Science and Technology, Wuhan, 430081, People’s Republic of China

    Jun Chen

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  1. Yongqing Liao
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  2. Min Xiong
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  5. Min Zhang
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  10. Jun Chen
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Contributions

YL: methodology, investigation, data curation, software, writing—original draft. MX: investigation, software. ZM: investigation, data curation. AI: writing—review and editing. MZ: investigation, writing—review and editing. BL: investigation, writing—review and editing. YZ: methodology, investigation, data curation. DC: methodology, investigation, data curation, writing—original draft. ZY: methodology, investigation, data curation. JC: data curation, writing—review and editing. SC: supervision, writing—review and editing.

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Correspondence to Jun Chen or Shouwen Chen.

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12010_2022_4206_MOESM1_ESM.doc

Supplementary file1 (DOC 1439 kb) The primers used in this research were provided in Table S1. The results of precursor amino acid additions on keratinase production, transcriptional level analysis of critical genes, and temperature stability experiments of keratinase were provided in Figure S1, S2 and S3, respectively. All these information could be attained in the additional file of this article.

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Liao, Y., Xiong, M., Miao, Z. et al. Modular Engineering to Enhance Keratinase Production for Biotransformation of Discarded Feathers. Appl Biochem Biotechnol 195, 1752–1769 (2023). https://doi.org/10.1007/s12010-022-04206-x

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