Abstract
We isolated the feather-degrading Bacillus pseudofirmus FA30-01 from the soil sample of poultry farm. The isolate completely degraded feather pieces after liquid culture at 30°C (pH 10.5) for 3 days. Strain FA30-01 is a Gram-positive, spore-forming, rod-shaped bacterium and was identified with B. pseudofirmus based on 16S rDNA analysis. The keratinase enzyme produced by strain FA30-01 was refined using ammonium sulfate precipitation, negative-ion DEAE Toyopearl exchange chromatography, and hydroxyapatite chromatography. The refinement level was 14.5-fold. The molecular weight of this enzyme was 27.5 kDa and it had an isoelectric point of 5.9. The enzyme exhibited activity at pH 5.1–11.5 and 30–80°C with azokeratin as a substrate, although the optimum pH and temperature for keratinase activity were pH 8.8–10.3 and 60°C, respectively. This enzyme is one of the serine-type proteases. Subtilisin ALP I and this enzyme had 90% homology in the N-terminal amino acid sequence. Since this enzyme differed from ALP I in molecular weight, heat resistance and isoelectric point, they are suggested to be different enzymes.
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Acknowledgements
Grant support from the Matsumoto farm is appreciated. We are grateful to Ayami Hideshima, Extremobiosphere Research Center, JAMSTEC, and Hiroaki Minegishi, Graduate School of Engineering, Toyo University, for 16SrDNA analysis. And we are also very grateful to Izumi Yoshikawa, Extremobiosphere Research Center, JAMSTEC, for NH2-terminal amino acids sequence.
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Kojima, M., Kanai, M., Tominaga, M. et al. Isolation and characterization of a feather-degrading enzyme from Bacillus pseudofirmus FA30-01. Extremophiles 10, 229–235 (2006). https://doi.org/10.1007/s00792-005-0491-y
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DOI: https://doi.org/10.1007/s00792-005-0491-y