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

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

Congreso; SiLaBB II - EnReBB VII; 2016

Diglycosidases are endo-β-glucosidases that hydrolyze the heterosidic linkage ofdiglycoconjugates, splitting off a disaccharide and the corresponding aglycone. Thefungus Acremonium sp. DSM 24697 produces at least two diglycosidases with differentsubstrate specificity. α-Rhamnosyl-β-glucosidase (ARBG) cleaves the disacchariderutinose from 7-O-rutinosylated flavonoids, while ARBGII cleaves rutinose from the3-O-rutinosylated flavonoid rutin (Fig 1).The genome of Acremonium sp. DSM 24697 was sequenced using the Illuminatechnique and assembled to 355 scaffolds (≥0.3 kb; N50, 235 kb) with a total size of ≈27Mb. ARBG and ARBGII were detected by zymographic assays and a tryptic digestionwas performed. The tryptic peptides were used as the probes for a BLAST searchagainst the translated genome. ARBG was predicted not to have introns, a theoreticalpI of 5.0 and a molecular mass of 41 kDa, while ARBG II was predicted to have threeintrons, 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 relationshipbetween sequence and substrate specificity of flavonoid-hydrolyzing enzymes.