In vitro and in vivo activation of L-serine deaminase in Escherichia coli K-12

Newman, E. B.; Dumont, Daniel; Walker, C

doi:10.1128/jb.162.3.1270-1275.1985

Cited by 19 publications

(13 citation statements)

References 27 publications

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“…Similarly, if E. coli K12 lost its serine deaminase activity in some other way, one would expect it also to filament in LB. To test this, we took advantage of an 'activation' mutant MEW191 in which the L-SD1 molecule is made and can be activated in vitro (Newman et al, 1985) but no active L-SD can be made in vivo. This mutant is functionally a triple mutant as none of the three enzymes can be activated, and indeed this strain also filamented on subculture into LB.…”

Section: Loss Of L-sd Activity Is Responsible For Filamentationmentioning

confidence: 99%

Deficiency in l‐serine deaminase results in abnormal growth and cell division of Escherichia coli K‐12

Zhang

Newman

2008

Molecular Microbiology

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SummaryThe loss of the ability to deaminate L-serine severely impairs growth and cell division in Escherichia coli K-12. A strain from which the three genes (sdaA, sdaB, tdcG) coding for this organism's three L-serine deaminases had been deleted grows well in glucose minimal medium but, on subculture into minimal medium with glucose and casamino acids, it makes very large, abnormally shaped cells, many of which lyse. When inoculated into Luria-Bertani (LB) broth with or without glucose, it makes very long filaments. Provision of S-adenosylmethionine restores cell division in LB broth with glucose, and repairs much of the difficulty in growth in medium with casamino acids. We suggest that replication of E. coli is regulated by methylation, that an unusually high intracellular L-serine concentration, in the presence of other amino acids, starves the cell for Sadenosylmethionine and that it is the absence of S-adenosylmethionine and/or of C1-tetrahydrofolate derivatives that prevents normal cell division.

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Section: Loss Of L-sd Activity Is Responsible For Filamentationmentioning

confidence: 99%

Deficiency in l‐serine deaminase results in abnormal growth and cell division of Escherichia coli K‐12

Zhang

Newman

2008

Molecular Microbiology

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“…Nevertheless, there would be a sufficient number of cysteinyl residues to complex at least one iron-sulfur cluster. In addition, L-serine dehydratase of E. coli is similarly activated by FeZ+ and dithiothreitol (Newman et al, 1985). Hence, there is ample reason to believe that the enzyme from E. coli might also be a member of the class of iron-sulfur-dependent enzymes.…”

Section: Discussionmentioning

confidence: 99%

“…These dehydratases are distinguished by their extreme instability and their ability to be protected, and in some cases activated, by substrate and/or competitive inhibitors (Alfoldi et al, 1968;Carter and Sagers, 1972;Morikawa et al, 1974;Gannon et al, 1977;Newman and Kapoor, 1980). Moreover, L-serine dehydratase from Clostridiuni acidi-urici (Carter and Sagers, 1972), E. coli (Newman et al, 1985). Lnctobacillus fermentum (Farias et al, 1991) and several other lactic acid bacteria (Farias et al, 1988) have been recognized to require activation by Fe".…”

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confidence: 99%

L‐Serine and L‐threonine dehydratase from Clostridium propionicum Two enzymes with different prosthetic groups

Hofmeister

Grabowski

Linder

1993

European Journal of Biochemistry

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L-Serine dehydratase from the Gram-positive bacterium Peptostreprococcus usuccharolyricus is novel in the group of enzymes deaminating 2-hydroxyamino acids in that it is an iron-sulfur protein and lacks pyridoxal phosphate [Grabowski, R. and Buckel, W. (1991) Eur. J. Biochem. 199, 89-941. It was proposed that this type of L-serine dehydratase is widespread among bacteria but has escaped intensive characterization due to its oxygen lability. Here, we present evidence that another Gram-positive bacterium, Clostridium pmpionicum, contains both an iron-sulfur-dependent L-serine dehydratase and a pyridoxal-phosphate-dependent L-threonine dehydratase. These findings support the notion that two independent mechanisms exist for the deamination of 2-hydroxyamino acids.L-Threonine dehydratase was purified 400-fold to apparent homogeneity and revealed as being a tetramer of identical subunits (m = 39 kDa). The purified enzyme exhibited a specific activity of 5 pkadmg protein and a K , for L-threonine of 7.7 mM. L-Serine ( K , = 380 mM) was also deaminated, the V/K, ratio, however, being 118-fold lower than the one for L-threonine. L-Threonine dehydratase was inactivated by borohydride, hydroxylamine and phenylhydrazine, all known inactivators of pyridoxal-phosphate-containing enzymes. Incubation with NaB3H, specifically labelled the enzyme. Activity of the phenylhydrazine-inactivated enzyme could be restored by pyridoxal phosphate.L-Serine dehydratase was also purified 400-fold, but its extreme instability did not permit purification to homogeneity. The enzyme was specific for L-serine ( K , = 5 mM) and was inhibited by L-cysteine (K, = 0.5 mM) and D-serine (K, = 8 mM). Activity was insensitive towards borohydride, hydroxylamine and phenylhydrazine but was rapidly lost upon exposure to air. Fez+ specifically reactivated the enzyme. L-Serine dehydratase was composed of two different subunits (a, m = 30kDa; p, m = 26kDa), their apparent molecular masses being similar to the ones of the two subunits of the iron-sulfur-dependent enzyme from Z? asuccharolyticus. Moreover, the N-terminal sequences of the small subunits from these two organisms were found to be 47% identical. In addition, 38% identity with the N-terminus of one of the two L-serine dehydratases of Escherichiu coli was detected.

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“…Most probably, the enzyme is only the first known member of a whole family of iron-sulfur-dependent L-serine dehydratases. There is evidence that enzymes from C. acidi-urici [88], lactic acid bacteria [89], E. coli [90] and C. propionicum (A.E.M. Hofmeister and W. Buckel, unpublished) also contain iron-sulfur clusters.…”

Section: L-serine Dehydratasementioning

confidence: 99%

Unusual dehydrations in anaerobic bacteria

Buckel

1992

FEMS Microbiology Letters

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In vitro and in vivo activation of L-serine deaminase in Escherichia coli K-12

Cited by 19 publications

References 27 publications

Deficiency in l‐serine deaminase results in abnormal growth and cell division of Escherichia coli K‐12

Deficiency in l‐serine deaminase results in abnormal growth and cell division of Escherichia coli K‐12

L‐Serine and L‐threonine dehydratase from Clostridium propionicum Two enzymes with different prosthetic groups

Unusual dehydrations in anaerobic bacteria

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