Aguiar, Tatiana Quinta

Domingues, Lucília

Heiskanen, Annamari

Maaheimo, Hannu

Penttilä, M.

Wiebe, M.

Universidade do Minho: RepositoriUM

bi ORGANIZED BY: !H=D ....................... 6th International Meeting on Biotechnology 19 - 21 September, 2012 - Bilbao (Spain) SCIENTIFIC CONGRESS BIOTEC 2012 SPONSOR: CO-SPONSOR: & y ~ EIAllcll-clolf'oll-~ ~..:-.,. ....... .,,=. - : hon Oral Communlutlons Session 8 S-8. OC1. Omics and Systems Biotechnology Profiling and structural analysis of Ashbya gossypti' N-Plvcans T. Q Aguiar1, H. Maaheimo2, A. Heiskanen3, M. Wiebe2, M. Penttilii2 and L. Domingues' '/BB - Insritute for Biorechnology and Bioengineering, Cenrre of Biological Engineering, Universidade do Mlnho. Campus de Gualrar. 4710-057 Braga. Portugal. /ucll/ad@deb.uminho.pr 'VTT Technical Research Centre of Finland, P.O. Box 1000, 02044 VTT Espoo, Finland 'Glykos Finland Lrd .. Helsinki, Finland Ashbyagossypilis a filamentous hemiascomycete used for the industrial production of riboflavin (vitamin 82) that has recently been explored as a new fungal host system for the expression and secretion of proteins. However, the secretory pathway of A. gossypii remains poorly characterized, although evidences indicate that the level of secretion in A. gossypii is more similar to yeast than to other filamentous fungi; and that recombrnant glycoproteins produced by A. gossypii are less extensively glycosylated than those produced by Saa:haromyces cerevisiae. A better understanding of the A. gossypll glycobiology offers a basis to characterize its protein secretory processes and may possibly be of practical impact in the production of heterologous proterns. As a starting point on the elucidation of A. gossypil N-glycosylation, in this study we performed a global analysis of whole mixtures of N-glycans attached to A. gossypii native secreted glycoproteins. Through combined analysis of N-glycan spectra obtained by MALDI-TOF mass spectrometry and 1 H NMR spectroscopy we report here that the major N-glycan species derived from A. gossypii secreted proteins are high-mannose type glycans containing core-type structures (Man8-9GlucNac2). Smaller core structures (Man5-7GlucNac2) are also present in lower abundance and N-glycans with long outer chains are only present In minor relative amounts. All identified acidic N-glycans contain a phosphate residue. Among the most abundant acidic structures are short phosphorylated outer chain glycans (PhMan 13-15GluNac2). Our data present the fi rst report on the composition, size distribution and structural characteristics of A. gossypii N-glycans. Acknowledgmencs: This work was supported by Fundat;ao para a C/encta e a Tecnologla (FCTJ. Portugal: Pro1ecr Ash8yofacrory gram PTDCJEBB· EBI/10198512008 and MIT-Portugal Program PhD grant SFRHIBD/3911212007 Co Tatt.ina Quintil Agular. The authors would like to thank Olli Aiclo for helping wlrh the lnterpreralion of the NMR specrra. 

Profiling and structural analysis of Ashbya gossypii N-glycans

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