Recombinant expression and molecular engineering of the keratinase from Brevibacillus parabrevis for dehairing performance

Rong-Xian Zhang, Jin Song Gong, Chang Su, Jiufu Qin, Heng Li, Hui Li, Jin Song Shi*, Zheng Hong Xu

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Keratinase is capable of distinctive degradation of keratin, which provides an eco-friendly approach for keratin waste management towards sustainable development. In this study, the recombinant keratinase (KERBP) from Brevibacillus parabrevis was successfully expressed in Escherichia coli. The purified KERBP had the specific activity of 6005.3 U/mg. It showed remarkable tolerance to various surfactants and also no collagenolytic activity. However, the moderate thermal stability limited its further application. Thus, protein engineering was further adopted to improve its stability. The variants of T218S, S236C and N181D were constructed by site-directed mutagenesis and combinatorial mutagenesis. Compared with the wild type, the t1/2 at 60 °C for the variants T218S, S236C and N181D were 3.05-, 1.18- and 1-fold increase, respectively. Moreover, the double variants N181D-T218S and N181D-S236C significantly improved thermostability with 5.1 and 2.9 °C increase of T50, and prolonging t1/2 at 60 °C with 4.09 and 1.54-fold, respectively. And the catalytic efficiency of the T218S and N181D-T218S variants was also significantly improved. Furthermore, the keratinase displayed favorable ability to dehair wool from skin within 7 h, which showed potential in leather dehairing. Our work contributes to a further insight into the thermostability of keratinase and offers a promising alternative for industrial leather application.
Original languageEnglish
JournalJournal of Biotechnology
Volume320
Pages (from-to)57-65
Number of pages9
ISSN0168-1656
DOIs
Publication statusPublished - 2020

Keywords

  • Brevibacillus parabrevis
  • Keratinase
  • Expression
  • Thermostability
  • Site-directed mutagenesis

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