Production and application of purified mutanase from novel Cellulosimicrobium funkei SNG1 in the in vitro biofilm degradation

Abstract

AbstractMutanase (α‐1,3‐glucanase) is an inducible extracellular enzyme with potential medical applications in dentistry. A novel Cellulosimicrobium funkei strain SNG1 producing mutanase enzyme using α‐1,3‐glucans was isolated, and the enzyme was optimized for increased productivity using the one‐factor‐at‐a‐time approach. Maximum growth and enzyme‐specific activity (2.12 ± 0.4 U/mg) were attained in a production medium with pH 7.0 and 1% α‐1,3‐glucans as carbon source, incubated at 37°C for 30 h. The result showed a five‐fold increase in activity compared to unoptimized conditions (0.40 U/mg). The enzyme was purified by gel‐filtration chromatography, and recovered with a yield of 29.03% and a specific activity increase of 10.9‐fold. The molecular mass of the monomeric enzyme is 137 kDa. The pH and temperature optima are 6.0 and 45°C with Km of 1.28 ± 0.11 mg for α‐1,3‐glucans. The enzyme activity was stimulated by adding Co2+, Ca2+, Cu2+, and was entirely inhibited by Hg2+. On 2‐h incubation, the purified enzyme effectively degraded in vitro film with an 82.68% degradation rate and a saccharification yield of 30%.