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Identification and Functional Characterization of a Fructooligosaccharides-Forming Enzyme from Aspergillus aculeatus

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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.

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Acknowledgments

This work was supported by the CONACyT, México (No. 169048) and PROMEP. José Juan Virgen-Ortíz would also like to express his gratitude to CONACyT Mexico for his PhD fellowship No. 307333.

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Authors and Affiliations

  1. Laboratorio de Bioingeniería, Universidad de Colima, Km. 9 carretera Coquimatlán-Colima, C.P. 28400, Coquimatlán, Colima, México

    José Juan Virgen-Ortíz, Vrani Ibarra-Junquera, Pilar Escalante-Minakata, Sara Centeno-Leija & Hugo Serrano-Posada

  2. Cátedras CONACyT, Dirección Adjunta de Desarrollo Científico, Consejo Nacional de Ciencia y Tecnología, Av. Insurgentes Sur 1582, 03940, México City, DF, México

    Sara Centeno-Leija & Hugo Serrano-Posada

  3. Centro de Investigación en Alimentación y Desarrollo A. C.-Unidad Cuauhtémoc, 31570, Cd. Cuauhtémoc, Chihuahua, México

    José de Jesús Ornelas-Paz

  4. Facultad de Ciencias Químicas, Universidad de San Luis Potosí, 78290, San Luis Potosí, México

    Jaime David Pérez-Martínez

  5. Facultad de Ciencias Biológicas y Agropecuarias, Universidad de Colima, 28100, Tecomán, Colima, México

    Juan Alberto Osuna-Castro

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  1. José Juan Virgen-Ortíz
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  2. Vrani Ibarra-Junquera
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Correspondence to Vrani Ibarra-Junquera.

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Virgen-Ortíz, J.J., Ibarra-Junquera, V., Escalante-Minakata, P. et al. Identification and Functional Characterization of a Fructooligosaccharides-Forming Enzyme from Aspergillus aculeatus . Appl Biochem Biotechnol 179, 497–513 (2016). https://doi.org/10.1007/s12010-016-2009-8

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