Creatinine and N-methylhydantoin degradation in two newly isolated Clostridium species

Abstract

With N-methylhydantoin (NMH) as the main organic substrate, two strictly anaerobic spore forming Gram-positive bacterial strains were isolated from sewage sludge. These strains, named Clostridium sp. FS23 and Clostridium sp. FS41, totally degraded NMH, via N-carbamoylsarcosine (CS) and sarcosine as intermediates. Strain FS23 grew also with creatinine, which was converted to NMH by creatinine iminohydrolase (EC 3.5.4.21). This enzyme was formed at high rates with all substrates tested. Cytosine and 5-fluorocytosine were not utilized as substrates by creatinine iminohydrolase preparations purified to a homogeneity of 98%. NMH amidohydrolase (NMHase) and N-carbamoylsarcosine amidohydrolase (CSHase) turned out to be inducible in both strains. Other than in aerobic organisms, NMHase from these two isolated did not require ATP for enzymatic activity. SH-group protecting agents were not necessary for stability.

The Phylogeny of the Genus Clostridium: Proposal of Five New Genera and Eleven New Species Combinations

“…Subcluster XIVa consisted of a phenotypically heterogeneous collection of organisms, including several non-spore-forming cocci (e.g., coprococci and ruminococci). Within subcluster XIVa, a biochemically unusual N-methylhydantoin-degrading mesophilic spore former (strain FS41 [= DSM 68771) isolated from sewage sludge (17) was found to be specifically related to Clostridium clostridiiforme. This unnamed strain (17) phenotypically resembles C. clostridiifome, but the observed level of sequence divergence (approximately 2.5%) indicates that this organism represents a new species.…”

Tissierella creatinophila sp. nov., a Gram-positive, anaerobic, non-spore-forming, creatinine-fermenting organism

“…Under anaerobic conditions the usual degradation pathway for creatinine involves the formation of Nmethylhydantoin, N-carbamoylsarcosine and sarcosine (Gauglitz, 1988;Hermann et al, 1992;Szulmajster, 1958a, b;ten Krooden & Owens, 1957). In contrast, the newly isolated strain KRE 4T degraded creatinine via creatine, sarcosine and glycine to the products acetyl phosphate, monomethylamine, ammonia and carbon dioxide.…”

“…Creatine amidinohydrolase is highly significant in medical diagnostics because creatinine clearance is used to monitor the filtration rate of glomeruli in the kidney (Siedel et al, 1988). Three metabolic pathways for the microbial degradation of creatinine have been described (Hermann et al, 1992). A strain classified as Arthrobacter ureafaciens was isolated which degraded creatinine via creatine and sarcosine and which was used in a bioassay for creatinine (Miller & Dubos, 1936 ;Kaplan & Naugler, 1974).…”

“…A strictly anaerobic, spore-forming, Gram-positive strain, Clostridium sp. FS23, which converted creatinine to N-methylhydantoin via a creatinine iminohydrolase has been isolated and characterized (Hermann et al, 1992).…”

Soehngenia saccharolytica gen. nov., sp. nov. and Clostridium amygdalinum sp. nov., two novel anaerobic, benzaldehyde-converting bacteria

“…C. hastiforme (Holdeman et al, 1977;Cato et al, 1982;Suen et al, 1988) is proteolytic, but not saccharolytic, unlike strain BOR-Y T . In addition to the traits shown in Table 1, Tissierella creatinini DSM 9508 T differs from strain BOR-Y T in that it grows on Nmethylhydantoin, and does not use carbohydrates or various other substrates including threonine (Gauglitz, 1988;Hermann et al, 1992), while Tissierella creatinophila (Harms et al, 1998) utilizes creatinine via creatine and does not grow on carbohydrates. Tissierella praeacuta (Tissier, 1908;Cato et al, 1979;Collins & Shah, 1986;Farrow et al, 1995) is Gram-negative, and is weakly or non-fermentative.…”