title: N-glycan modification in Aspergillus species.
abstract: The production by filamentous fungi of therapeutic glycoproteins intended for use in mammals is held back by the inherent difference in protein N-glycosylation and by the inability of the fungal cell to modify proteins with mammalian glycosylation structures. Here, we report protein N-glycan engineering in two Aspergillus species. We functionally expressed in the fungal hosts heterologous chimeric fusion proteins containing different localization peptides and catalytic domains. This strategy allowed the isolation of a strain with a functional alpha-1,2-mannosidase producing increased amounts of N-glycans of the Man5GlcNAc2 type. This strain was further engineered by the introduction of a functional GlcNAc transferase I construct yielding GlcNAcMan5GlcNac2 N-glycans. Additionally, we deleted algC genes coding for an enzyme involved in an early step of the fungal glycosylation pathway yielding Man3GlcNAc2 N-glycans. This modification of fungal glycosylation is a step toward the ability to produce humanized complex N-glycans on therapeutic proteins in filamentous fungi.
authors: Fungal Genomics Unit, Austrian Research Centers and BOKU Vienna, Muthgasse 18, A-1190 Vienna, Austria.
authors: Elke
authors: E
authors: Kainz
authors: Andreas
authors: A
authors: Gallmetzer
authors: Christian
authors: C
authors: Hatzl
authors: Juergen H
authors: JH
authors: Nett
authors: Huijuan
authors: H
authors: Li
authors: Thorsten
authors: T
authors: Schinko
authors: Robert
authors: R
authors: Pachlinger
authors: Harald
authors: H
authors: Berger
authors: Yazmid
authors: Y
authors: Reyes-Dominguez
authors: Andreas
authors: A
authors: Bernreiter
authors: Tillmann
authors: T
authors: Gerngross
authors: Stefan
authors: S
authors: Wildt
authors: Joseph
authors: J
authors: Strauss
mesh_terms: D001230:Aspergillus / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018014:Gene Transfer Techniques; D008364:Mannosyltransferases / Q000235:genetics; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000096:biosynthesis* / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014169:Transformation, Bacterial / Q000235:genetics*; D043323:alpha-Mannosidase / Q000378:metabolism
chemical_list: D017931:DNA Primers; D011134:Polysaccharides; D011993:Recombinant Fusion Proteins; D008364:Mannosyltransferases; D043323:alpha-Mannosidase