Isolation of dicarboxylic acid- and glucose-binding proteins from Pseudomonas aeruginosa

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Abstract
Inducible binding proteins for C4-dicarboxylic acids (DBP) and glucose (GBP) were isolated from P. aeruginosa by extraction of exponential-phase cells with 0.2 M MgCl2 (pH 8.5) and by an osmotic shock procedure without affecting cell viability. DBP synthesis was induced by growth on aspartate, .alpha.-ketoglutarate, succinate, fumarate, malate and malonate but not by growth on acetate, citrate, pyruvate or glucose. Binding of succinate by DBP was competitively inhibited by 10-fold concentrations of fumarate and malate but not by a variety of related substances. GBP synthesis and transport of methyl .alpha.-glucoside by whole cells were induced by growth on glucose or pyruvate plus galactose, 2-deoxyglucose or methyl .alpha.-glucoside but not by growth on gluconate, succinate, acetate or pyruvate. The binding of radioactive glucose by GBP was significantly inhibited by 10-fold concentrations of glucose, galactose and glucose-1-phosphate but not by the other carbohydrates tested. The binding of glucose by GBP or succinate by DBP did not result in any chemical alteration of the substrates.