Trehalose 6-phosphate synthase from Dictyostelium discoideum: Partial purification and characterization of the enzyme from young sorocarps

Killick, Kathleen A.

doi:10.1016/0003-9861(79)90558-7

Cited by 26 publications

(14 citation statements)

References 22 publications

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“…The abundant cellulose in this medium suggests cellulase may be involved; synthesis of trehalose from the hexose produced by the action of a cellulase could be mediated by trehalose phosphate synthetase, as is found for Dictyostelium discoideum (Killick, 1979). Although the latter enzyme has not been established in A. bisporus, an extracellular endocellulase, which exhibits a coincidence and correlation with flushing similar to that found above for trehalase and glycogen phosphorylase, has been demonstrated (Wood & Goodenough, 1977;Manning & Wood, 1983;N.…”

Section: Discussionmentioning

confidence: 52%

VARIATIONS IN ACTIVITIES OF GLYCOGEN PHOSPHORYLASE AND TREHALASE DURING THE PERIODIC FRUITING OF THE EDIBLE MUSHROOM AGARICUS BISPORUS (LANGE) IMBACH.

1987

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SUMMARYActivities of trehalase and glycogen phosphorylase were determined for stage 1 (pin) sporophores of Agaricus bisporus harvested daily throughout several fruiting cycles (flushes). Both enzymes exhibited maximum activity coincident with peak flush, as defined by maximum yield of stage 2 to 4 sporopbores and minimum activity during the interflush periods. Glycogen phosphorylase activity sometimes peaked on more than one occasion during a flush, whereas trehalase gave only a single peak of activity per flush. These results are discussed in relation to metabolic events associated with carbobydrate changes during tbe periodic fruiting cycle of tbis fungus.

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Section: Discussionmentioning

confidence: 52%

VARIATIONS IN ACTIVITIES OF GLYCOGEN PHOSPHORYLASE AND TREHALASE DURING THE PERIODIC FRUITING OF THE EDIBLE MUSHROOM AGARICUS BISPORUS (LANGE) IMBACH.

1987

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“…In cell extracts, the OtsA of P. freudenreichii was absolutely specific for ADP-glucose, a finding that appears to be unique as far as OtsA proteins are concerned. In fact, known OtsA proteins either can use the four glucosyl donors (Lapp et al, 1971;Killick, 1979;Pan et al, 2002;Silva et al, 2005), or are specific for UDP-or GDPglucose (Elbein, 1968;Giaever et al, 1988). Most surprising was the observation that the pure (native or recombinant) OtsA of P. freudenreichii lacked the absolute specificity shown in cell extracts, and recognized any of the four sugar-nucleotides as a substrate.…”

Section: Discussionmentioning

confidence: 99%

Biochemical and genetic characterization of the pathways for trehalose metabolism in Propionibacterium freudenreichii, and their role in stress response

2007

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Propionibacterium freudenreichii accumulates high levels of trehalose, especially in response to stress. The pathways for trehalose metabolism were characterized, and their roles in response to osmotic, oxidative and acid stress were studied. Two pathways were identified: the trehalose-6-phosphate synthase/phosphatase (OtsA-OtsB) pathway, and the trehalose synthase (TreS) pathway. The former was used for trehalose synthesis, whereas the latter is proposed to operate in trehalose degradation. The activities of OtsA, OtsB and TreS were detected in cell extracts; the corresponding genes were identified, and the recombinant proteins were characterized in detail. In crude extracts of P. freudenreichii, OtsA was specific for ADP-glucose, in contrast to the pure recombinant OtsA, which used UDP-, GDP-and TDP-glucose, in addition to ADP-glucose. Moreover, the substrate specificity of OtsA in cell extracts was lost during purification, and the recombinant OtsA became specific to ADP-glucose upon incubation with a dialysed cell extract. The level of OtsA was enhanced (approximately twofold) by osmotic, oxidative and acid stress, whereas the level of TreS remained constant, or it decreased, under identical stress conditions. Therefore, the OtsA-OtsB pathway plays an important role in the synthesis of trehalose in response to stress. It is most likely that trehalose degradation proceeds via TreS to yield maltose, which is subsequently catabolized via amylomaltase activity. Hydrolytic activities that are potentially involved in trehalose degradation (trehalase, trehalose phosphorylase, trehalose-6-phosphate phosphorylase and trehalose-6-phosphate hydrolase) were not present. The role of trehalose as a common response to three distinct stresses is discussed.

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“…This disaccharide has also been reported in yeast (1,5), in fungi (12,22), as well as in bacteria (3), mainly in actinomycetes (15). In soybean plants trehalose appears to be restricted to the nodule tissue (23).…”

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confidence: 93%

Enzymes of α,α-Trehalose Metabolism in Soybean Nodules

Salminen¹,

Streeter²

1986

Plant Physiol.

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Metabolism of trehalose, a,D-glucopyranosyl-a,D-glucopyranoside, was studied in nodules of Bradyrhizobium japonicum-Glycine max IL.I Merr. cv Beeson 80 symbiosis. The nodule extract was divided into three fractions: bacteroid soluble protein, bacteroid fragments, and cytosol. The bacteroid soluble protein and cytosol fractions were gel-filtered. The key biosynthetic enzyme, trehalose-6-phosphate synthetase, was consistently found only in the bacteroids. Trehalose-6-phosphate phosphatase activity was present both in the bacteroid soluble protein and cytosol fractions. Trehalase, the most abundant catabolic enzyme was present in all three fractions and showed two pH optima: pH 3.8 and 6.6. Two other degradative enzymes, phosphotrehalase, acting on trehalose-6-phosphate forming glucose and glucose-6-phosphate, and trehalose phosphorylase, forming glucose and 0-glucose-l-phosphate, were also detected in the bacteroid soluble protein and cytosol fractions. Trehalase was present in large excess over trehalose-6-phosphate synthetase. Trehalose accumulation in the nodules would appear to be predicated on spatial separation of trehalose and trehalase.Trehalose, a-D-glucopyranosyl-a-D-glucopyranoside, is one of the major carbohydrates synthesized by cultured Bradyrhizobium japonicum, and every species of Rhizobium examined so far shows trehalose accumulation (26). This disaccharide has also been reported in yeast (1,5), in fungi (12, 22), as well as in bacteria (3), mainly in actinomycetes (15). In soybean plants trehalose appears to be restricted to the nodule tissue (23). Its presence elsewhere in higher plants has not been conclusively established (7). Previous work with soybean nodules suggests that trehalose is synthesized by the microsymbiont (20).The pathway for trehalose synthesis was first elucidated in Saccharomyces cerevisiae (5) (Fig. 1). The biosynthetic enzymes have also been reported in Lilium longiflorum pollen (8).There appears to be more than one way of catabolizing trehalose (Fig. 1). Phosphotrehalase has been reported in Bacillus popilliae (3) and trehalose phosphorylase in Euglena gracilis (17). Trehalase, the most widely occurring catabolic enzyme has also been found in higher plants (6,9,27 Nodule Fractionation. After 5 to 6 weeks the plants were harvested, and subsequently all operations were conducted at 0 to 2°C. Five g of nodules were ground in a mortar with a pestle in 10 ml (added in 2 ml aliquots) of grinding medium, 0.1 M sodium phosphate buffer (pH 7.5) containing 0.15 M mannitol, 2 mm DTE, and 1 mm EDTA. The homogenate was filtered through two Miracloth discs in a 50 ml syringe into a 15 ml centrifuge tube.The Miracloth filtrate was then centrifuged at 5OOg for 10 min. The resultant starch pellet was discarded and the supernatant fluid recentrifuged at 6,000g for 10 min to obtain the bacteroid pellet. The supernatant fluid was centrifuged at 48,000g for 15 min and the supernatant, cytosol fraction, was saved for gel filtration.The bacteroid pellet was resuspended in 8 ml of grindin...

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Trehalose 6-phosphate synthase from Dictyostelium discoideum: Partial purification and characterization of the enzyme from young sorocarps

Cited by 26 publications

References 22 publications

VARIATIONS IN ACTIVITIES OF GLYCOGEN PHOSPHORYLASE AND TREHALASE DURING THE PERIODIC FRUITING OF THE EDIBLE MUSHROOM AGARICUS BISPORUS (LANGE) IMBACH.

VARIATIONS IN ACTIVITIES OF GLYCOGEN PHOSPHORYLASE AND TREHALASE DURING THE PERIODIC FRUITING OF THE EDIBLE MUSHROOM AGARICUS BISPORUS (LANGE) IMBACH.

Biochemical and genetic characterization of the pathways for trehalose metabolism in Propionibacterium freudenreichii, and their role in stress response

Enzymes of α,α-Trehalose Metabolism in Soybean Nodules

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