Characterization of a Mannose-6-Phosphate Isomerase from Bacillus amyloliquefaciens and Its Application in Fructose-6-Phosphate Production

• Published in: PloS one Vol. 10; no. 7; p. e0131585
• Main Authors: Sigdel, Sujan, Singh, Ranjitha, Kim, Tae-Su, Li, Jinglin, Kim, Sang-Yong, Kim, In-Won, Jung, Woo-Suk, Pan, Cheol-Ho, Kang, Yun Chan, Lee, Jung-Kul
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 14.07.2015; Public Library of Science (PLoS)
• Subjects: Amino Acid Sequence; Bacillus - enzymology; Bacillus - genetics; Bacillus amyloliquefaciens; Bacillus licheniformis; Bacillus subtilis; Candida albicans; Catalysis; Chemical engineering; Chemical synthesis; Cloning, Molecular; Crystallography, X-Ray; Dehydrogenases; E coli; Enzymatic activity; Enzyme activity; Enzyme Stability; Enzymes; Escherichia coli - genetics; Fructose; Fructose-6-phosphate; Fructosephosphates - biosynthesis; Functional foods & nutraceuticals; Hydrogen-Ion Concentration; Industrial applications; Kinetics; Mannose; Mannose-6-phosphate isomerase; Mannose-6-Phosphate Isomerase - chemistry; Mannose-6-Phosphate Isomerase - genetics; Mannose-6-Phosphate Isomerase - metabolism; Metabolism; Metals - pharmacology; Molecular Docking Simulation; Molecular Dynamics Simulation; Molecular Sequence Data; Molecular Weight; pH effects; Phosphate; Phosphates; Protein Structure, Quaternary; Salmonella; Salmonella Typhimurium; Sequence Homology, Amino Acid; Studies; Substrate Specificity; Substrates; Sugar; Temperature; Zinc
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0131585

The BaM6PI gene encoding a mannose-6-phosphate isomerase (M6PI, EC 5.3.1.8) was cloned from Bacillus amyloliquefaciens DSM7 and overexpressed in Escherichia coli. The enzyme activity of BaM6PI was optimal at pH and temperature of 7.5 and 70°C, respectively, with a kcat/Km of 13,900 s-1 mM-1 for mannose-6-phosphate (M6P). The purified BaM6PI demonstrated the highest catalytic efficiency of all characterized M6PIs. Although M6PIs have been characterized from several other sources, BaM6PI is distinguished from other M6PIs by its wide pH range and high catalytic efficiency for M6P. The binding orientation of the substrate M6P in the active site of BaM6PI shed light on the molecular basis of its unusually high activity. BaM6PI showed 97% substrate conversion from M6P to fructose-6-phosphate demonstrating the potential for using BaM6PI in industrial applications.