Characterization of glycerol kinase and NAD-independent glycerol-3-phosphate dehydrogenase from Pediococcus pentosaceus N5p

Abstract

S A AD . 1998. The pathway involved in glycerol dissimilation in Pediococcus pentosaceus N5p, a strain isolated from wine, includes glycerol kinase and NAD-independent glycerol-3P dehydrogenase. The properties of these enzymes were studied. Glycerol kinase activity was maximal at 28°C and pH 7·5 in 50 mmol l −1 TrisHCl buffer. The end-products of the reaction acted as competitive inhibitors while fructose-1,6-diphosphate was a non-competitive inhibitor. Mg 2+ was required for optimal enzyme activity. The Km values for both substrates were 0·11 and 0·37 mmol l −1 for glycerol and ATP, respectively. NAD-independent glycerol-3P dehydrogenase activity was maximal at 37°C and pH 7·5 in 100 mmol l −1 Tris-HCl buffer. The enzymatic activity was activated by KCN and bivalent cations as Mg 2+ and Ca 2+, but it was strongly inhibited by others. Dihydroxyacetone phosphate acted as competitive inhibitor while ATP and phosphenolpyruvate were non-competitive inhibitors.

Glycerol Metabolism of <i>Lactobacillus rhamnosus</i> ATCC 7469: Cloning and Expression of Two Glycerol Kinase Genes

“…A previous work reported that L. casei Cog 812 had the ability to use glycerol as a sole energy source [Lonvaud-Funel, 1986]. A Pediococcus pentosaceus strain showed a similar behavior using glycerol as sole energy source producing D-lactate, acetate and in certain conditions diacetyl and acetoin [Pasteris and Strasser de Saad, 1998]. …”

Expression, purification, and characterization of a His6-tagged glycerokinase from Pichia farinosa for enzymatic cycling assays in mammalian cells

“…The K m value of P. farinosa GK for ATP was generally higher than those of other GKs. While the K ATP was estimated to be approximately 428 M, the enzymes from Pyrococcus kodakaraensis, T. brucei, and P. pentosaceus had values ranging from 15 to 370 M (Koga et al, 1998;Krakow and Wang, 1990;Pasteris and de Saad, 1998). Besides, the GK from E. coli (Novotny et al, 1985), Haemophilius influenzae (Pawlyk and Pettigrew, 2001) and (Fig.…”

“…5B). Previous studies indicated that the temperature optimum ranged from 25 to 80 • C for GK from different species (Hayashi and Lin, 1967;Koga et al, 1998;Pasteris and de Saad, 1998).…”

“…This reaction was characterized and compared over a broad range of species like bacteria (e.g. Escherichia coli, Pediococcus pentosaceus) (Pasteris and de Saad, 1998;Thorner and Paulus, 1973), yeasts (e.g. Debaryomyces hansenii, Candida mycoderma) (Janson and Cleland, 1974;Nilsson et al, 1989), and animals (e.g.…”

Characterization of glycerol kinase from baker's yeast: Response surface modeling of the enzymatic reaction

“…In addition, the enzyme is industrially important and useful for the clinical determination of the blood triglyceride level in combination with lipase, glycerol-3-phosphate oxidase and peroxidase (PO) [1]. Up to now, GKs have been purified from several microorganisms [2][3][4][5][6][7] and a vertebrate [8], and Abbreviations: GK, glycerol kinase; GPO, glycerol phosphate oxidase; PO, peroxidase; RSM, response surface methodology; R 2 , determination coefficient (quadratic correlation coefficient); R 2 adj , adjusted R 2 . * Corresponding author.…”