Formation of glucosylceramide and sterol glucoside by a UDP-glucose-dependent glucosylceramide synthase from cotton expressed in Pichia pastoris |
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Authors: | Hillig Inga Leipelt Martina Ott Claudia Zähringer Ulrich Warnecke Dirk Heinz Ernst |
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Institution: | Institut für Allgemeine Botanik, University of Hamburg, Ohnhorststr. 18, 22609 Hamburg, Germany. |
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Abstract: | In plants, glucosylceramide (GlcCer) biosynthesis is poorly understood. Previous investigations suggested that sterol glucoside (SG) acts as the actual glucose donor for the plant GlcCer synthase (GCS). We addressed this question by generating a Pichia pastoris double mutant devoid of GlcCer and SG. This mutant was used for heterologous expression of the plant GCS. The activity of the GCS resulted in the accumulation of GlcCer and, surprisingly, a small proportion of SG. The synthesis of GlcCer in the transformed double mutant shows that the GCS is SG-independent, while the detection of SG suggests that in addition to the sterol glucosyltransferase, also the GCS may contribute in planta to SG biosynthesis. |
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Keywords: | Cerebroside Glucosylceramide synthase Glycosphingolipid Sterol glucoside Pichia pastoris |
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