title: Identification and Functional Characterization of a Fructooligosaccharides-Forming Enzyme from Aspergillus aculeatus.
abstract: Although fructosyltransferases from Aspergillus aculeatus have received a considerable interest for the prebiotics industry, their amino acid sequences and structural features remain unknown. This study sequenced and characterized a fructosyltransferase from A. aculeatus (AcFT) isolated by heat treatment of Pectinex Ultra SP-L. The AcFT enzyme showed two isoforms, low-glycosylated AcFT1 and high-glycosylated AcFT2 forms, with similar optimum activity at 60 °C. The purified heat-resistant AcFT1 and AcFT2 isoforms produced identical patterns of fructooligosaccharides (FOS; kestose, nystose and fructosylnystose) with a notable transfructosylation capability (~90 % transferase/hydrolase ratio). In contrast, the pI and optimum pH values exhibited discrete differences, attributable to their glycosylation pattern. Partial protein sequencing showed that AcFT enzyme corresponds to Aspac1_37092, a putative 654-residue fructosyltransferase encoded in the genome of A. aculeatus ATCC16872. A homology model of AcFT also revealed the typical fold common to members of the glycoside hydrolase family 32 (GH32), with an N-terminal five-blade β-propeller domain enclosing catalytic residues D60, D191, and E292, linked to a C-terminal β-sandwich domain. To our knowledge, this is the first report describing the amino acid sequence and structural features of a heat-resistant FOS-forming enzyme from A. aculeatus, providing insights into its potential applications in the prebiotics industry.
authors: Laboratorio de Bioingeniería, Universidad de Colima, Km. 9 carretera Coquimatlán-Colima, C.P. 28400, Coquimatlán, Colima, México.
authors: José Juan
authors: JJ
authors: Virgen-Ortíz
authors: Laboratorio de Bioingeniería, Universidad de Colima, Km. 9 carretera Coquimatlán-Colima, C.P. 28400, Coquimatlán, Colima, México. vij@ucol.mx.
authors: Vrani
authors: V
authors: Ibarra-Junquera
authors: Laboratorio de Bioingeniería, Universidad de Colima, Km. 9 carretera Coquimatlán-Colima, C.P. 28400, Coquimatlán, Colima, México.
authors: Pilar
authors: P
authors: Escalante-Minakata
authors: Laboratorio de Bioingeniería, Universidad de Colima, Km. 9 carretera Coquimatlán-Colima, C.P. 28400, Coquimatlán, Colima, México.|Cátedras CONACyT, Dirección Adjunta de Desarrollo Científico, Consejo Nacional de Ciencia y Tecnología, Av. Insurgentes Sur 1582, 03940, México City, DF, México.
authors: Sara
authors: S
authors: Centeno-Leija
authors: Laboratorio de Bioingeniería, Universidad de Colima, Km. 9 carretera Coquimatlán-Colima, C.P. 28400, Coquimatlán, Colima, México.|Cátedras CONACyT, Dirección Adjunta de Desarrollo Científico, Consejo Nacional de Ciencia y Tecnología, Av. Insurgentes Sur 1582, 03940, México City, DF, México.
authors: Hugo
authors: H
authors: Serrano-Posada
authors: Centro de Investigación en Alimentación y Desarrollo A. C.-Unidad Cuauhtémoc, 31570, Cd. Cuauhtémoc, Chihuahua, México.
authors: José
authors: J
authors: de Jesús Ornelas-Paz
authors: Facultad de Ciencias Químicas, Universidad de San Luis Potosí, 78290, San Luis Potosí, México.
authors: Jaime David
authors: JD
authors: Pérez-Martínez
authors: Facultad de Ciencias Biológicas y Agropecuarias, Universidad de Colima, 28100, Tecomán, Colima, México.
authors: Juan Alberto
authors: JA
authors: Osuna-Castro
mesh_terms: D000595:Amino Acid Sequence; D001230:Aspergillus / Q000737:chemistry / Q000201:enzymology*; D001483:Base Sequence; D006602:Hexosyltransferases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D007700:Kinetics; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000737:chemistry; D056692:Prebiotics; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D001426:Bacterial Proteins
chemical_list: D006602:Hexosyltransferases; D009844:Oligosaccharides; D056692:Prebiotics; C116580:fructooligosaccharide; C021451:inulosucrase; D001426:Bacterial Proteins
keywords: Amino acid sequence; Fructosyltransferase; Heat treatment; Transfructosylation