Browsing by Author "Ya’u S. Ajingi"

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    Identification, overexpression, purification, and biochemical characterization of a novel hyperthermostable keratinase from Geoglobus acetivorans
    (2021-01-01) Duangjai Sittipol; Sudarat Rodpan; Ya’u S. Ajingi; Tassanee Lohnoo; Tassanee Lerksuthirat; Yothin Kumsang; Wanta Yingyong; Pongsak Khunrae; Triwit Rattanarojpong; Kovit Pattanapanyasat; Nujarin Jongruja; Ramathibodi Hospital; Siriraj Hospital; Kano University of Science and Technology; King Mongkut's University of Technology Thonburi
    The goal of this study was to identify and biochemically characterize a novel hyperthermostable keratinase from microorganisms for feather waste degradation. Here, a hyperthermophilic Geoglobus acetivorans keratinase (GacK) gene was chosen based on a search of a sequence database. The selected GacK gene was synthesized, cloned, and successfully expressed without a signal peptide in the E. coli system. A monomer of approximately 58 kDa was obtained in a soluble form and purified. The recombinant GacK displayed the highest activity at an optimum temperature of 100 °C and a pH of 10. The hyperthermostable GacK enzymatic performance remained high even after incubation in nonionic surfactants and the chelating agent EDTA. The residual and keratinolytic activities of GacK, as determined with azocasein and keratin azure used as substrates, remained significantly greater than 80% at 130 °C for 7 h. The kinetic parameters Km and Vmax for azure keratin were 0.41 mg/ml and 875.14 unit/mg, respectively, while those for azocasein were 1.51 mg/ml and 505.32 unit/mg, respectively. The results suggest that the enzyme is among the most hyperthermostable keratinases. Because of its enzymatic characteristics to degrade keratin azure at high temperatures, GacK may potentially be utilized in future industrial applications.