Relation between Germination, Trehalose and the Status of Water in Phycomyces blakesleeanus Spores as Measured by Proton-NMR

Laere, André Van; François, A; Overloop, K; Verbeke, Marc N.; Gerven, L. Van

doi:10.1099/00221287-133-2-239

Cited by 10 publications

(9 citation statements)

References 23 publications

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“…Spores of S. venezuelae, but not the mycelium, are resistant to dehydration (Glazebrook et al 1990). In Phycomyces blakesleeanus trehalose has been reported to protect spores from desiccation damage (Van Laere et al 1987) and to have a specific role in preserving membrane structure in the dry state (Crowe et al 1984). Evidence for a similar role in S. antibioticus has been presented by Martin et al (1986), and trehalose may well play a role in preserving S. venezuelae spores during dormancy.…”

Section: Discussionmentioning

confidence: 95%

Accumulation of intracellular carbon reserves in relation to chloramphenicol biosynthesis by Streptomyces venezuelae

Ranade

Vining²

1993

Can. J. Microbiol.

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Two chloramphenicol-producing strains of Streptomyces venezuelae accumulated small amounts of polyhydroxybutyrate during exponential growth; the compound disappeared from the mycelium as the cultures entered stationary phase. Depletion of polyhydroxybutyrate coincided with chloramphenicol production but the amount of polymer stored in the mycelium was insufficient to supply the precursor requirement for biosynthesis of the antibiotic. Accumulation of polyhydroxybutyrate in the S. venezuelae strains was appreciably lower than in two other streptomycetes examined. Glycogen and lipids accumulated in the mycelium of S. venezuelae 13s during the stationary phase, after nitrogen depletion; under the culture conditions used, they were the principal storage compounds in S. venezuelae. Trehalose was absent from the mycelium in vegetative cultures grown under nonsporulating conditions but it was abundant in spores obtained from submerged and surface cultures. Glycogen and polyhydroxybutyrate were absent from spores.

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Section: Discussionmentioning

confidence: 95%

Accumulation of intracellular carbon reserves in relation to chloramphenicol biosynthesis by Streptomyces venezuelae

Ranade

Vining²

1993

Can. J. Microbiol.

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“…The free trehalose in dormant M. smegmatis cells observed in this study is reminiscent of the accumulation of trehalose in large amounts in dormant yeast ascospores, actinomycete spores, and in fungal spores (Arguelles, 2000; Elbein et al, 2003). As suggested, trehalose decreases the mobility of cytoplasmic water resulting in global down-regulation of enzyme activity and spore polymer stabilization providing their prolonged viability (Van Laere et al, 1987). Another possible role of trehalose in dormant mycobacteria might be its usage as a carbohydrate store as proposed for yeast and filamentous fungal spores (Thevelein, 1984).…”

Section: Discussionmentioning

confidence: 99%

Free Trehalose Accumulation in Dormant Mycobacterium smegmatis Cells and Its Breakdown in Early Resuscitation Phase

et al. 2017

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Under gradual acidification of growth medium resulting in the formation of dormant Mycobacterium smegmatis, a significant accumulation of free trehalose in dormant cells was observed. According to 1H- and 13C-NMR spectroscopy up to 64% of total organic substances in the dormant cell extract was represented by trehalose whilst the trehalose content in an extract of active cells taken from early stationary phase was not more than 15%. Trehalose biosynthesis during transition to the dormant state is provided by activation of genes involved in the OtsA-OtsB and TreY-TreZ pathways (according to RT-PCR). Varying the concentration of free trehalose in dormant cells by expression of MSMEG_4535 coding for trehalase we found that cell viability depends on trehalose level: cells with a high amount of trehalose survive much better than cells with a low amount. Upon resuscitation of dormant M. smegmatis, a decrease of free trehalose and an increase in glucose concentration occurred in the early period of resuscitation (after 2 h). Evidently, breakdown of trehalose by trehalase takes place at this time as a transient increase in trehalase activity was observed between 1 and 3 h of resuscitation. Activation of trehalase was not due to de novo biosynthesis but because of self-activation of the enzyme from the inactive state in dormant cells. Because, even a low concentration of ATP (2 mM) prevents self-activation of trehalase in vitro and after activation the enzyme is still sensitive to ATP we suggest that the transient character of trehalase activation in cells is due to variation in intracellular ATP concentration found in the early resuscitation period. The negative influence of the trehalase inhibitor validamycin A on the resuscitation of dormant cells proves the importance of trehalase for resuscitation. These experiments demonstrate the significance of free trehalose accumulation for the maintenance of dormant mycobacterial viability and the involvement of trehalose breakdown in early events leading to cell reactivation similar to yeast and fungal spores.

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“…In some other fungi the mechanism of regulation remains uncertain (20). Dormant fungal spores have a low water content (21). The water content of Phycomyces blakesleeanus spores increases during germination at about the same time that trehalose mobilization occurs.…”

Section: Discussionmentioning

confidence: 99%

Regulation of trehalose metabolism by Streptomyces griseus spores

McBride

Ensign

1990

J Bacteriol

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Spores of Streptomyces griseus contain trehalose and trehalase, but trehalose is not readily hydrolyzed until spore germination is initiated. Trehalase in crude extracts of spores, germinated spores, and mycelia of S. griseus had a pH optinum of approximately 6.2, had a Km value for trehalose of approximately 11 mM, and was most active in buffers having ionic strengths of 50 to 200 mM. Inhibitors or activators or trehalase activity were not detected in extracts of spores or mycelia. Several lines of evidence indicated that trehalose and trehalase are both located in the spore cytoplasm. Spores retained their trehalose and most of their trehalase activity following brief exposure to dilute acid. Protoplasts formed by enzymatic removal of the spore walls in buffer containing high concentrations of solutes also retained their trehalose and trehalase activity. Protoplasts formed in buffer containing lower levels of solutes contained low levels of trehalose. The mechanism by which trehalose metabolism is regulated in S. griseus spores is unresolved. A low level of hydration of the cytoplasm of the dormant spores and an increased level of hydration during germination may account for the apparent inactivity of trehalase in dormant spores and the rapid hydrolysis of trehalose upon initiation of germination.Large amounts of the disaccharide trehalose are present in the spores and cysts of a variety of organisms, including actinomycetes, fungi, nematodes, and brine shrimp (2, 4, 13). In these organisms trehalose is degraded very slowly during periods of dormancy but is rapidly metabolized following the onset of vegetative growth (1,9,14,20,22).Spores of Streptomyces griseus contain large amounts of trehalose (13). Extracts of the spores also contain a high specific activity of the enzyme trehalase (14). Nongerminating spores metabolize their endogenous trehalose reserves very slowly. Upon transfer to conditions allowing spore germination, trehalose is rapidly metabolized, while the level of trehalase activity remains constant (14).In this paper we report the results of an investigation of the properties of trehalase and the localization of trehalase activity and trehalose in spores of S. griseus. The goal of this study was to determine the basis for the apparent coexistence of trehalase and trehalose in dormant spores and the rapid utilization of trehalose following the initiation of germination.MATERIALS AND METHODS Growth conditions. S. griseus NRRL B-2682 was maintained as described previously (13). Spores were harvested from solid DMC medium after 7 days of incubation at 30°C unless indicated otherwise. DMC medium was modified to obtain lysozyme-sensitive spores by adding glycine to a final concentration of 0.35% and by adjusting the pH to 6.6 prior to sterilization. Spores were germinated in the complex germination medium described previously (13). Mycelia were grown in liquid DMC medium containing 0.15 M KCl. Protoplast regeneration medium consisted of 25 mM morpholinopropanesulfonic acid (MOPS) (pH 7.0), 5 mM (NH4)...

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Relation between Germination, Trehalose and the Status of Water in Phycomyces blakesleeanus Spores as Measured by Proton-NMR

Cited by 10 publications

References 23 publications

Accumulation of intracellular carbon reserves in relation to chloramphenicol biosynthesis by Streptomyces venezuelae

Accumulation of intracellular carbon reserves in relation to chloramphenicol biosynthesis by Streptomyces venezuelae

Free Trehalose Accumulation in Dormant Mycobacterium smegmatis Cells and Its Breakdown in Early Resuscitation Phase

Regulation of trehalose metabolism by Streptomyces griseus spores

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