RESUMEN

ArbZ from Lactobacillus delbrueckii subsp. lactis was previously shown to enable utilization of the beta-glucoside arbutin by Escherichia coli. The arbZ gene was cloned and expressed in the industrially used beta-glucoside-negative strain Lactobacillus helveticus 3036(62). The transformants were able to ferment not only arbutin, but also cellobiose, salicin and methyl-beta-glucoside (MbetaGlc). Cleavage of beta-glucosides by the transformants depended on the integrity of the cytoplasmic membrane, whereas in cell-free extracts only C(6)-phosphorylated substrates were hydrolysed. This suggested that ArbZ is a phospho-beta-glycosidase. ArbZ activity in transformants of Lb. helveticus was subject to substrate induction mediated by the beta-glucosides arbutin, salicin and MbetaGlc, whereas cellobiose or the beta-galactoside lactose had no inducing effect. Northern blot analysis proved that induction by MbetaGlc was due to enhanced transcription of arbZ. Catabolite repression of arbZ induction was observed with glucose, mannose, fructose and galactose. The induction kinetics observed in the presence of these sugars indicated that at least two different mechanisms are operative in catabolite repression of arbZ in Lb. helveticus.

Asunto(s)

Proteínas Bacterianas/biosíntesis , Glicósido Hidrolasas/biosíntesis , Lactobacillus/enzimología , Arbutina/metabolismo , Proteínas Bacterianas/genética , Secuencia de Bases , Clonación Molecular , Cartilla de ADN/genética , Inducción Enzimática/efectos de los fármacos , Fermentación , Genes Bacterianos , Glucósidos/metabolismo , Glucósidos/farmacología , Glicósido Hidrolasas/genética , Lactobacillus/efectos de los fármacos , Lactobacillus/genética , Transformación Genética