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To elucidate the regulation and limiting factors in the glycosylation of secreted proteins, the mpg1 and dpm1 genes from Trichoderma reesei (Hypocrea jecorina) encoding GTP:alpha-D-mannose-1-phosphate guanyltransferase and dolichyl phosphate mannose synthase (DPMS), respectively, were overexpressed in T. reesei. No significant increases were observed in DPMS activity or protein secretion in dpm1-overexpressing transformants, whereas overexpression of mpg1 led to a twofold increase in GDP-mannose (GDPMan) levels. GDPMan was effectively utilized by mannnosyltransferases and resulted in hypermannosylation of secreted proteins in both N and O glycosylation. Overexpression of the mpg1 gene also increased the transcription of the dpm1 gene and DPMS activity. Our data indicate that the level of cellular GDPMan can play a major regulatory role in protein glycosylation in T. reesei.

applied and environmental microbiology

Overexpression of the Gene Encoding GTP:Mannose-1-Phosphate Guanyltransferase, <i>mpg1</i> , Increases Cellular GDP-Mannose Levels and Protein Mannosylation in <i>Trichoderma reesei</i>

Overexpression of the Gene Encoding GTP:Mannose-1-Phosphate Guanyltransferase, mpg1 , Increases Cellular GDP-Mannose Levels and Protein Mannosylation in Trichoderma reesei