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Applied and Environmental Microbiology, December 2006, p. 7778-7784, Vol. 72, No. 12
0099-2240/06/$08.00+0     doi:10.1128/AEM.02375-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Glycoprotein Hypersecretion Alters the Cell Wall in Trichoderma reesei Strains Expressing the Saccharomyces cerevisiae Dolichylphosphate Mannose Synthase Gene{triangledown}

Urszula Perlinska-Lenart,1 Jacek Orlowski,1 Agnieszka E. Laudy,2 Ewa Zdebska,3 Grazyna Palamarczyk,1 and Joanna S. Kruszewska1*

Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland,1 Department of Pharmaceutical Microbiology, Medical University, Oczki 3, 02-007 Warsaw, Poland,2 Department of Biochemistry, Institute of Hematology and Blood Transfusion, Chocimska 5, 00-957 Warsaw, Poland3

Received 9 October 2006/ Accepted 10 October 2006

Expression of the Saccharomyces cerevisiae DPM1 gene (coding for dolichylphosphate mannose synthase) in Trichoderma reesei (Hypocrea jecorina) increases the intensity of protein glycosylation and secretion and causes ultrastructural changes in the fungal cell wall. In the present work, we undertook further biochemical and morphological characterization of the DPM1-expressing T. reesei strains. We established that the carbohydrate composition of the fungal cell wall was altered with an increased amount of N-acetylglucosamine, suggesting an increase in chitin content. Calcofluor white staining followed by fluorescence microscopy indicated changes in chitin distribution. Moreover, we also observed a decreased concentration of mannose and alkali-soluble ß-(1,6) glucan. A comparison of protein secretion from protoplasts with that from mycelia showed that the cell wall created a barrier for secretion in the DPM1 transformants. We also discuss the relationships between the observed changes in the cell wall, increased protein glycosylation, and the greater secretory capacity of T. reesei strains expressing the yeast DPM1 gene.

* Corresponding author. Mailing address: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland. Phone: 48 22 592 12 09. Fax: 48 22 658 46 36. E-mail: jsk{at}ibb.waw.pl.

{triangledown} Published ahead of print on 20 October 2006.

Applied and Environmental Microbiology, December 2006, p. 7778-7784, Vol. 72, No. 12
0099-2240/06/$08.00+0     doi:10.1128/AEM.02375-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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