1887

Abstract

This is a study of high-affinity glucose uptake in and the effect of disruption of a high-affinity monosaccharide-transporter gene, . The substrate saturation constant ( ) of a reference strain was about 15 μM in glucose-limited chemostat culture. Disruption of resulted in a two- to fivefold reduction in affinity for glucose and led to expression of a low-affinity glucose transport gene, , at high dilution rate. The effect of disruption was more subtle at low and intermediate dilution rates, pointing to some degree of functional redundancy in the high-affinity uptake system of . The disruptant and a reference strain were cultivated in glucose-limited chemostat cultures at low, intermediate and high dilution rate (=0.07 h, 0.14 h and 0.20 h). Mycelium harvested from steady-state cultures was subjected to glucose uptake assays, and analysed for expression of and two other transporter genes, and . The capacity for glucose uptake ( ) of both strains was significantly reduced at low dilution rate. The glucose uptake assays revealed complex uptake kinetics. This impeded accurate determination of maximum specific uptake rates ( ) and apparent affinity constants () at intermediate and high dilution rate. Two high-affinity glucose transporter genes, and , were expressed at all three dilution rates in chemostat cultures, in contrast to batch culture, where only was expressed. Expression patterns of the three transporter genes suggested differential regulation and functionality of their products.

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2007-06-01
2020-04-10
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