The phosphoinositide dependent kinase PDK1 activates the serum- and glucocorticoid-inducible kinase and protein kinase B isoforms which in turn are known to stimulate the renal and intestinal Na⁺-dependent glucose transporter SGLT1. The present study has been performed to explore the role of PDK1 in electrogenic glucose transport in small intestine and proximal renal tubules. To this end, mice expressing ~20 % of PDK1 (pdk1^hm) were compared to their wild type littermates (pdk1^wt). According to Ussing chamber experiments electrogenic glucose transport was significantly smaller in the jejunum of pdk1^hm than of pdk1^wt mice. Similarly, proximal tubular electrogenic glucose transport in isolated perfused renal tubule segments was decreased in pdk1^hm as compared to pdk1^wt mice. Intraperitoneal injection of 3 g/kg bw glucose resulted in a similar increase of plasma glucose concentration in pdk1^hm and in pdk1^wt mice but led to a higher increase of urinary glucose excretion in pdk1^hm mice. In conclusion, reduction of functional PDK1 leads to impairment of electrogenic intestinal glucose absorption and renal glucose reabsorption. The experiments disclose a novel element of glucose transport regulation in kidney and small intestine.