title: In Silico Analysis of the Structure of Fungal Fructooligosaccharides-Synthesizing Enzymes.
abstract: Fructooligosaccharides (FOS) are prebiotics commonly manufactured using fungal fructosyltransferases (FTases) or β-fructofuranosidases. Several reports have attempted to optimize FOS production by changing operational conditions. Nevertheless, there is a lack of information related to the molecular enzyme-substrate interaction. In this study, we present an in silico evaluation of the interactions between substrates (i.e., glucose, sucrose, GF2, GF3, and GF4) and native FOS-synthesizing enzymes from fungi, with reported FOS production yield. In addition, a molecular dynamic simulation was conducted to assess the stability of these interactions. Six fungal enzymes with reported data of FOS production were selected: sucrose-sucrose 1-fructosyltransferase from A. foetidus (GenBank No. CAA04131); intracellular invertase from A. niger (GenBank No. ABB59679); extracellular invertase from A. niger (GenBank No. ABB59678); β-fructofuranidase from A. japonicus ATCC 20611 (GenBank No. BAB67771); fructosyltransferase from A. oryzae N74 (GenBank No. ACZ48670); and fructosyltransferase from A. japonicus (PDB ID 3LF7). These enzymes shared an identity between 15 and 96 %, but have a highly conserved folding, and the characteristic FTases domains. Docking results showed that these enzymes also share a similar protein-ligand interaction profile. It was observed that the production yield of total FOS correlated with the sum of affinity energies for GF2, GF3, and GF4. Finally, we present the first molecular dynamic simulation for FOS and fungal FOS-synthesizing enzymes, showing that the protein-ligand interaction does not induce significant changes on the enzyme stability. Overall, these results represent valuable information to continue understanding the FOS synthesis process by fungal FOS-synthesizing enzymes, and they can have a significant impact toward the improvement in their catalytic properties and the synthesis of specific FOS.
authors.affiliation: Clinical Bacteriology Program, School of Health Sciences, Universidad Colegio Mayor de Cundinamarca, Bogotá, Colombia.|Chemical Department, School of Science, Pontificia Universidad Javeriana, Bogotá, Colombia.
authors.forename: Sergio
authors.initials: S
authors.lastname: Olarte-Avellaneda
authors.affiliation: Chemical Department, School of Science, Pontificia Universidad Javeriana, Bogotá, Colombia.|Institute for the Study of Inborn Errors of Metabolism, Pontificia Universidad Javeriana, Kra 7 No. 43-82 Building 54, Room 303A, Bogotá, Colombia.
authors.forename: Alexander
authors.initials: A
authors.lastname: Rodríguez-López
authors.affiliation: Institute for the Study of Inborn Errors of Metabolism, Pontificia Universidad Javeriana, Kra 7 No. 43-82 Building 54, Room 303A, Bogotá, Colombia.
authors.forename: Juan David
authors.initials: JD
authors.lastname: Patiño
authors.affiliation: Institute for the Study of Inborn Errors of Metabolism, Pontificia Universidad Javeriana, Kra 7 No. 43-82 Building 54, Room 303A, Bogotá, Colombia. cjalmeciga@javeriana.edu.co.
authors.forename: Carlos J
authors.identifier: http://orcid.org/0000-0001-6484-1173
authors.initials: CJ
authors.lastname: Alméciga-Díaz
authors.affiliation: School of Chemical Engineering, Purdue University, Forney Hall of Chemical Engineering, 480 Stadium Mall Drive, West Lafayette, IN, 47907-2100, USA. sancheo@purdue.edu.
authors.forename: Oscar F
authors.initials: OF
authors.lastname: Sánchez
mesh_terms: D000595:Amino Acid Sequence; D001230:Aspergillus / Q000201:enzymology*; D003198:Computer Simulation; D005656:Fungal Proteins / Q000737:chemistry; D006860:Hydrogen Bonding; D007700:Kinetics; D008024:Ligands; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D016297:Mutagenesis, Site-Directed; D009844:Oligosaccharides / Q000096:biosynthesis*
chemical_list: D005656:Fungal Proteins; D008024:Ligands; D009844:Oligosaccharides; C116580:fructooligosaccharide
keywords: Aspergillus; Fructooligosaccharides; Fructosyltransferases; Molecular docking; Molecular dynamics