Isolation of the GFA1 gene encoding glucosamine-6-phosphate synthase of Sporothrix schenckii and its expression in Saccharomyces cerevisiae - Publication - Bridge of Knowledge

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Isolation of the GFA1 gene encoding glucosamine-6-phosphate synthase of Sporothrix schenckii and its expression in Saccharomyces cerevisiae

Abstract

Glucosamine-6-phosphate synthase (GlcN-6-P synthase) is an essential enzyme involved in cell wall biogenesis that has been proposed as a strategic target for antifungal chemotherapy. Here we describe the cloning and functional characterization of Sporothrix schenckii GFA1 gene which was isolated from a genomic library of the fungus. The gene encodes a predicted protein of 708 amino acids that is homologous to GlcN-6-P synthases from other sources. The recombinant enzyme restored glucosamine prototrophy of the Saccharomyces cerevisiae gfa1 null mutant. Purification and biochemical analysis of the recombinant enzyme revealed some differences from the wild type enzyme, such as improved stability and less sensitivity to UDP-GlcNAc. The sensitivity of the recombinant enzyme to the selective inhibitor FMDP [N3-(4- methoxyfumaroyl)-L-2,3-diaminopropanoic acid] and other properties were similar to those previously reported for the wild type enzyme.

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Category:
Articles
Type:
artykuł w czasopiśmie wyróżnionym w JCR
Published in:
PROTEIN EXPRESSION AND PURIFICATION no. 110, pages 57 - 64,
ISSN: 1046-5928
Language:
English
Publication year:
2015
Bibliographic description:
Sánchez-López J., Gonzalez-Ibarra J., Álvarez-Vargas A., Milewski S., Villagomez-Castro J., Cano-Canchola C., Lopez-Romero E.: Isolation of the GFA1 gene encoding glucosamine-6-phosphate synthase of Sporothrix schenckii and its expression in Saccharomyces cerevisiae// PROTEIN EXPRESSION AND PURIFICATION. -Vol. 110, (2015), s.57-64
DOI:
Digital Object Identifier (open in new tab) 10.1016/j.pep.2014.12.002
Verified by:
Gdańsk University of Technology

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