We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Purification and characterization of an intracellular levansucrase derived from Bacillus methylotrophicus SK 21.002.
- Authors
Li, Runjing; Zhang, Tao; Jiang, Bo; Mu, Wanmeng; Miao, Ming
- Abstract
An intracellular levansucrase from Bacillus methylotrophicus SK 21.002 was isolated, purified, and characterized. The final specific levansucrase activity was 135.40 U/mg protein with an 11.78-fold enrichment and a 9.28% recovery rate. The molecular weight of the enzyme was approximately 60,000 Da as evaluated by gel filtration and SDS-PAGE. Both the maximum transfructosylation and hydrolytic activities were observed at pH 6.5. The enzyme exhibited optimum transfructosylation activity at 40°C, whereas the optimum temperature of hydrolytic activity was 45°C. Cu2+, Fe2+, Zn2+, and Ni2+ inhibited both the transfructosylation and hydrolytic activities up to 100%, whereas Mn2+ inhibited only hydrolytic activity. Ca2+ and Mg2+ stimulated both transfructosylation and hydrolytic activities. The chemical modifiers (n-bromosuccinimide and phenylmethanesulfonyl fluoride) strongly inhibited hydrolytic and transfructosylation activity of the levansucrase. The Km and Vmax values of the purified levansucrase were 117.2 mM and 33.23 µmol/mg⋅Min, respectively. When the fructose concentration was below 0.2 M, higher fructose concentrations promoted the transfructosylation and inhibited the hydrolytic activity.
- Subjects
LEVANSUCRASE; BACILLUS genetics; MOLECULAR weights; HYDROLASES; CHEMICAL modification of proteins
- Publication
Biotechnology & Applied Biochemistry, 2015, Vol 62, Issue 6, p815
- ISSN
0885-4513
- Publication type
Article
- DOI
10.1002/bab.1334