Glycogen and Trehalose Accumulation during Colony Development in Streptomyces antibioticus

Af, Braña; Méndez, Cármen; La, Díaz; Mb, Manzanal; Hardisson, Carlos

doi:10.1099/00221287-132-5-1319

Cited by 36 publications

(36 citation statements)

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“…On the other hand, Glc-1P is the initial metabolite of routes allowing sugar/NDP-sugar interconversion, synthesis of oligoand polysaccharide structural components, or production of reserve glucans (glycogen) (12,32). A critical point determining metabolic fates for Glc-1P/Glc-6P within a cell is the presence or absence of UDP-sugar PPase, an enzyme that so far has been found in plants and protozoa (15,16,22,26,28,29,41).…”

Section: Figmentioning

confidence: 99%

Characterization of Recombinant UDP- and ADP-Glucose Pyrophosphorylases and Glycogen Synthase To Elucidate Glucose-1-Phosphate Partitioning into Oligo- and Polysaccharides in Streptomyces coelicolor

et al. 2012

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b Streptomyces coelicolor exhibits a major secondary metabolism, deriving important amounts of glucose to synthesize pigmented antibiotics. Understanding the pathways occurring in the bacterium with respect to synthesis of oligo-and polysaccharides is of relevance to determine a plausible scenario for the partitioning of glucose-1-phosphate into different metabolic fates. We report the molecular cloning of the genes coding for UDP-and ADP-glucose pyrophosphorylases as well as for glycogen synthase from genomic DNA of S. coelicolor A3(2). Each gene was heterologously expressed in Escherichia coli cells to produce and purify to electrophoretic homogeneity the respective enzymes. UDP-glucose pyrophosphorylase (UDP-Glc PPase) was characterized as a dimer exhibiting a relatively high V max in catalyzing UDP-glucose synthesis (270 units/mg) and with respect to dTDP-glucose (94 units/mg). ADP-glucose pyrophosphorylase (ADP-Glc PPase) was found to be tetrameric in structure and specific in utilizing ATP as a substrate, reaching similar activities in the directions of ADP-glucose synthesis or pyrophosphorolysis (V max of 0.15 and 0.27 units/mg, respectively). Glycogen synthase was arranged as a dimer and exhibited specificity in the use of ADP-glucose to elongate ␣-1,4-glucan chains in the polysaccharide. ADP-Glc PPase was the only of the three enzymes exhibiting sensitivity to allosteric regulation by different metabolites. Mannose-6-phosphate, phosphoenolpyruvate, fructose-6-phosphate, and glucose-6-phosphate behaved as major activators, whereas NADPH was a main inhibitor of ADP-Glc PPase. The results support a metabolic picture where glycogen synthesis occurs via ADP-glucose in S. coelicolor, with the pathway being strictly regulated in connection with other routes involved with oligo-and polysaccharides, as well as with antibiotic synthesis in the bacterium. Gram-positive bacteria of the genus Streptomyces exhibit a particularly complex morphological differentiation, with the formation of a highly branched substrate and aerial mycelium that generates spores from septation of the aerial hyphae (23, 33). Also characteristic of Streptomyces spp. is the occurrence of a flush secondary metabolism, with these bacteria producing almost twothirds of all known natural antibiotics (9). Streptomyces coelicolor A3(2) is the producer of three pigmented antibiotics, and it has a well-characterized genetics; these characteristics make it one of the preferred strains for research within this genus of bacteria. Although the complete genome of this S. coelicolor strain was elucidated about 10 years ago (5), advanced knowledge of the microorganism at the biochemical level is far from being reached. Most of the studies carried out with Streptomyces spp. have been biased toward the understanding of its morphological and physiological differentiation, while very little has been done on the biochemistry of the enzymes involved in the metabolic pathways.Monosaccharide interconversion and synthesis of oligo-and polysaccharides are key proces...

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Section: Figmentioning

confidence: 99%

Characterization of Recombinant UDP- and ADP-Glucose Pyrophosphorylases and Glycogen Synthase To Elucidate Glucose-1-Phosphate Partitioning into Oligo- and Polysaccharides in Streptomyces coelicolor

et al. 2012

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“…DISCUSSION Trehalose is present in large quantities in S. griseus and is most abundant in spores in which it may enhance spore resistance (1,4,(16)(17)(18).…”

Section: Resultsmentioning

confidence: 99%

“…Nongerminating spores of Streptomyces viridochromogenes, Streptomyces antibioticus, and Micromonospora echinospora and nongrowing spherical cells of Arthrobacter crystalopoietes and Nocardia coralina also maintained large amounts of trehalose and active trehalase. These trehalose reserves were depleted during spore germination or outgrowth of spherical Arthrobacter and Nocardia cells into rods.Large amounts of the disaccharide trehalose are present in the dormant spores or cysts of a variety of organisms including actinomycetes, fungi, nematodes, and brine shrimp (1,3,5,6,18). Trehalose is degraded very slowly by these organisms during periods of dormancy but is rapidly metabolized following the onset of vegetative growth (2,11,20,22).…”

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Metabolism of endogenous trehalose by Streptomyces griseus spores and by spores or cells of other actinomycetes

McBride

Ensign

1987

J Bacteriol

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The disaccharide trehalose is accumulated as a storage product by spores of Streptomyces griseus. Nongerminating spores used their trehalose reserves slowly when incubated in buffer for several months. In contrast, spores rapidly depleted their trehalose pools during the first hours of germination. Extracts of dormant spores contained a high specific activity of the enzyme trehalase. The level of trehalase remained relatively constant during germination or incubation in buffer. Nongerminating spores of Streptomyces viridochromogenes, Streptomyces antibioticus, and Micromonospora echinospora and nongrowing spherical cells of Arthrobacter crystalopoietes and Nocardia coralina also maintained large amounts of trehalose and active trehalase. These trehalose reserves were depleted during spore germination or outgrowth of spherical Arthrobacter and Nocardia cells into rods.Large amounts of the disaccharide trehalose are present in the dormant spores or cysts of a variety of organisms including actinomycetes, fungi, nematodes, and brine shrimp (1,3,5,6,18). Trehalose is degraded very slowly by these organisms during periods of dormancy but is rapidly metabolized following the onset of vegetative growth (2,11,20,22).We previously reported (15, 18) the accumulation of large amounts of trehalose by spores of Streptomyces griseus and the effects of trehalose content on spore properties. In this report we describe the results of an investigation of the metabolism of the endogenous reserves of trehalose by dormant and germinating spores of S. griseus. Results of studies of the metabolism of trehalose by some other actinomycetes are also described. MATERIALS AND METHODSGrowth conditions. S. griseus NRRL B-2682 spores were harvested from DMC agar as described previously (18 Germination conditions for S. griseus spores. The complex germination medium contained 0.5% yeast extract and 0.5% casein hydrolysate in 10 mM 3-morpholinepropanesulfonic acid (MOPS) buffer (pH 7.0). The defined germination medium consisted of 10 mM potassium phosphate buffer (pH 7.0)-5 mM (NH4)2SO4-0.5 mM MgSO4-7.6 mM Lisoleucine-7.6 mM L-leucine-8.5 mM L-valine-4.9 mM Ltryptophan-0.75 mM adenosine-14 mM glucose. In some experiments glucose was replaced by 27 mM glycerol. Spores were heat activated (15 min at 45°C) in 10 mM glycine buffer (pH 9.5), rapidly cooled to 0°C, and washed 3 times with 10 mM potassium phosphate buffer (pH 7.0) by centrifugation. Activated spores were inoculated into the germination media at final concentrations of approximately 2 x 108 spores per ml. The spores were incubated in 100 ml of medium in 1-liter flasks at 33°C with shaking at 185 rpm.Germination was monitored microscopically and was quantitated by taking advantage of the relative resistance of dormant spores to dilute acid. The number of acid-resistant CFUs was used to measure the number of dormant spores during sporulation or germination. Samples (0.2 ml) were added to 0.8 ml of 0.125 N HCI, dispersed by gentle sonication in a sonic water bath (Heat Systems Ultrasonics...

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“…The content of trehalose in aerial spores was higher than in the submerged spores. It has been reported that trehalose is the major carbohydrate in Streptomyces spores (Ensign, 1978;Brana et al, 1986), and also that the resistance of actinomycete spores to heat and desiccation is linked closely to their content of trehalose (Kendrick & Ensign, 1983;Dworkin, 1985;McBride & Ensign, 1987a, b).…”

Section: Discussionmentioning

confidence: 99%

Characteristics of spores formed by surface and submerged cultures of Streptomyces albidoflavus SMF301

Lee¹,

Rho²

1993

Journal of General Microbiology

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Stveptomyces albidoflavus SMF301 produced abundant spores in submerged cultures (submerged spores) as well as on solid media (aerial spores). The content of carbon, hydrogen, nitrogen, and phosphorus in submerged and aerial spores was similar; however, the contents of metal ions (K+, Na+, Ca2+ and Mg2+) were very different. Glutamic acid, alanine, and glycine, all known to be cell-wall components, were the major amino acids in both types of spores. However, cysteine was more abundant in submerged spores than in aerial spores. The major fatty acid in aerial spores was n-C,, (61.74%), whereas in submerged spores it was ai-C,, (33.68%). The contents of ai-C,,, and ai-C,, in submerged spores were also very much higher than in aerial spores. Unsaturated fatty acids were found in both kinds of spores but not in mycelium; they were particularly abundant in submerged spores. The composition of menaquinones in the two kinds of spores also varied. The resistance of aerial spores to lysozyme digestion, mild acid treatment, heating and desiccation was higher than that of submerged spores, but the submerged spores were more resistant to sonication.

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Glycogen and Trehalose Accumulation during Colony Development in Streptomyces antibioticus

Cited by 36 publications

References 20 publications

Characterization of Recombinant UDP- and ADP-Glucose Pyrophosphorylases and Glycogen Synthase To Elucidate Glucose-1-Phosphate Partitioning into Oligo- and Polysaccharides in Streptomyces coelicolor

Characterization of Recombinant UDP- and ADP-Glucose Pyrophosphorylases and Glycogen Synthase To Elucidate Glucose-1-Phosphate Partitioning into Oligo- and Polysaccharides in Streptomyces coelicolor

Metabolism of endogenous trehalose by Streptomyces griseus spores and by spores or cells of other actinomycetes

Characteristics of spores formed by surface and submerged cultures of Streptomyces albidoflavus SMF301

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