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• W3164244039 endingPage "5774" @default.
• W3164244039 startingPage "5766" @default.
• W3164244039 abstract "Levan-type fructooligosaccharides (LFOs) and levan can potentially be used as ingredients in prebiotics, skincare products, and antitumor agents. The Y246S mutant of Bacillus licheniformis RN-01 levansucrase (oligosaccharide-producing levansucrase, OPL) was reported to productively synthesize LFOs; however, OPL's thermostability is low at high temperatures. To enhance OPL structural stability, this study employed molecular dynamics (AMBER) to identify a highly flexible region, as measured by its average root-mean-square fluctuation (RMSF) value, on the OPL surface and computational protein design (Rosetta) to rigidify and increase favorable interactions to increase its structural stability. AMBER identified region nine (residues 277-317) as a highly flexible region that was selected for design because it has the highest number of residues and the second-highest average RMSF, and it is farthest from the active site. Rosetta designed 14 mutants with the best ΔΔG value in each position, where three mutants have better ΔG than OPL. To determine whether their flexibilities and stabilities are lower than those of OPL, all 14 designed mutants were simulated at high temperature (500 K), and we found that K296E, G309S, and A310W mutants were predicted to be more stable and could retain their native structures better than OPL. Our results suggest that enhanced structural stabilities of these mutants are caused by increased hydrogen bond strengths of the designed residues and their neighboring residues. This study designed K296E, G309S, and A310W mutants of OPL with high potential for stability improvement, and they could potentially be used for the effective production of LFOs." @default.
• W3164244039 created "2021-06-07" @default.
• W3164244039 creator A5036888888 @default.
• W3164244039 creator A5083564868 @default.
• W3164244039 creator A5049780611 @default.
• W3164244039 date "2021-05-28" @default.
• W3164244039 modified "2023-10-07" @default.
• W3164244039 title "Computational Design of Oligosaccharide-Producing Levansucrase from <i>Bacillus licheniformis</i> RN-01 to Increase Its Stability at High Temperature" @default.
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• W3164244039 doi "https://doi.org/10.1021/acs.jpcb.1c02016" @default.
• W3164244039 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/34047564" @default.
• W3164244039 hasPublicationYear "2021" @default.
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