The genome of Acremonium sp. DSM 24697 codes for two diglycosidases with different substrate specificity

GISELA WEIZ; BARBARA D NEHER; LAURA S. MAZZAFERRO; MICHAEL KOTIK; PETR HALADA; VLADIMIR KREN; JAVIER D. BRECCIA

Simposio; II Simposio Latinoamericano de Biocatálisis y Biotransformaciones (SiLaBB II) VII Encuentro Regional de Biocatálisis y Biotransformaciones; 2016

Universidad de la República

Diglycosidases are endo-β-glucosidases that hydrolyze the heterosidic linkage of diglycoconjugates, splitting off a disaccharide and the corresponding aglycone. The fungus Acremonium sp. DSM 24697 produces at least two diglycosidases with different substrate specificity. α-Rhamnosyl-β-glucosidase (ARBG) cleaves the disaccharide rutinose from 7-O-rutinosylated flavonoids, while ARBGII cleaves rutinose from the 3-O-rutinosylated flavonoid rutin (Fig 1).The genome of Acremonium sp. DSM 24697 was sequenced using the Illumina technique and assembled to 355 scaffolds (≥0.3 kb; N50, 235 kb) with a total size of ≈27 Mb. ARBG and ARBGII were detected by zymographic assays and a tryptic digestion was performed. The tryptic peptides were used as the probes for a BLAST search against the translated genome. ARBG was predicted not to have introns, a theoretical pI of 5.0 and a molecular mass of 41 kDa, while ARBG II was predicted to have three introns, a theoretical pI of 4.5 and a molecular mass of 80.8 kDa.The cloning and expression of these genes will bring insights into the relationship between sequence and substrate specificity of flavonoid-hydrolyzing enzymes.