Sudip Regmi, Seung W. Kim, Jin C. Yoo, Yoon S. Choi, Yun H. Choi, Hah Y. Yoo
Mannan-degrading enzymes have been growing interest in bio-industrial applications, such as the pulp and paper, food, and pharmaceutical industries. In this study, an extremely alkaline mannanase (MnB31) is produced by Bacillus subtilis subsp. inaquosorum CSB31. MnB31 is purified to 17.92-fold with a 21.51% yield and specific activity of 1,796.13 U mg−1 by anion-exchange and gel filtration column chromatography. The biochemical characterization of MnB31 is performed, and the results are as follows: molecular weight of ≈47 kDa with an optimum temperature of 60 °C and pH of 12.5. The enzyme is strongly activated by Co2+, Mn2+, Na+, and K+, and inhibited by Zn2+, Ni2+, and Mg2+. Halo-tolerance (10% NaCl), urea stability (3 M), and protease resistance are also observed. The kinetic parameters of MnB31 are found to be Km of 0.043 mg ml−1, and Vmax of 1,046 ± 3.605 U mg−1, respectively. In addition, the thermodynamical parameters are investigated; the activation energy (Ea) is found to be 31.36 kJ mol−1 with a Kcat value of 156.9 × 104 s−1, ΔH (28.59 kJ mol−1), ΔG (42.38 kJ mol−1), ΔS (−41.39 J mol−1 K−1), Q10 (1.40), ΔGE–S (−8.697 kJ mol−1), and ΔGE-T (−48.22 kJ mol−1). These results suggest that MnB31 has potential bio-industrial application, due to its greater hydrolytic efficiency and feasibility of enzymatic reaction.
An extremely alkaline mannanase (MnB31) is produced by Bacillus subtilis subsp. inaquosorum CSB31. MnB31 was purified by anion-exchange and gel filtration column chromatography. The biochemical characterization of MnB31 is performed, and the kinetic parameters and thermodynamical parameters are investigated. The formation of major end products such as mannobiose and mannotriose in the enzymatic hydrolysis is highly useful as prebiotics in the food and pharmaceutical industries. This article is part of an AFOB (Asian Federation of Biotechnology) Special issue. To learn more about the AFOB visit www.afob.org.