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- Title
Expression, characterization, and activity optimization of a novel cellulase from the thermophilic bacteria Cohnella sp. A01.
- Authors
Mohammadi, Shima; Tarrahimofrad, Hossein; Arjmand, Sareh; Zamani, Javad; Haghbeen, Kamahldin; Aminzadeh, Saeed
- Abstract
Cellulases are hydrolytic enzymes with wide scientific and industrial applications. We described a novel cellulase, CelC307, from the thermophilic indigenous Cohnella sp. A01. The 3-D structure of the CelC307 was predicted by comparative modeling. Docking of CelC307 with specific inhibitors and molecular dynamic (MD) simulation revealed that these ligands bound in a non-competitive manner. The CelC307 protein was purified and characterized after recombinant expression in Escherichia coli (E. coli) BL21. Using CMC 1% as the substrate, the thermodynamic values were determined as Km 0.46 mM, kcat 104.30 × 10–3 (S−1), and kcat/Km 226.73 (M−1 S−1). The CelC307 was optimally active at 40 °C and pH 7.0. The culture condition was optimized for improved CelC307 expression using Plackett–Burman and Box–Behnken design as follows: temperature 20 °C, pH 7.5, and inoculation concentration with an OD600 = 1. The endoglucanase activity was positively modulated in the presence of Na+, Li+, Ca2+, 2-mercaptoethanol (2-ME), and glycerol. The thermodynamic parameters calculated for CelC307 confirmed its inherent thermostability. The characterized CelC307 may be a suitable candidate for various biotechnological applications.
- Subjects
THERMOPHILIC bacteria; HYDROLASES; CELLULASE; ESCHERICHIA coli; MERCAPTOETHANOL
- Publication
Scientific Reports, 2022, Vol 12, Issue 1, p1
- ISSN
2045-2322
- Publication type
Article
- DOI
10.1038/s41598-022-14651-7