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

Abstract

A strictly anaerobic, Gram-positive, non-spore-forming bacterium was isolated from sewage sludge which grew on creatinine as sole source of carbon and energy. This new isolate, designated strain KRE 4l, totally degraded creatinine via creatine, sarcosine and glycine to the products acetate, monomethylamine, ammonia and carbon dioxide. Growth on creatinine or creatine was seleniumdependent and stimulated by formate, indicating the involvement of a creatine reductase, sarcosine reductase and/or glycine reductase. This was substantiated by the fact that creatine, sarcosine and glycine were reduced by cell-free extracts. Growth on creatinine or creatine was also possible in the absence of formate, but with an increase in doubling time. The new bacterium occurred as rod-shaped cells, which exhibited an angular form (2-6 pm long and 0.7-1.1 pm wide) and showed motility by means of peritrichous flagella. The G+C content of the DNA was 30 mol0/o. Comparative 16s rRNA sequence analysis demonstrated that strain KRE 4T represents a new subline within the genus Tissierella. Due to its very restricted substrate spectrum and the inability of whole cells to utilize sarcosine and glycine as intermediates of creatine breakdown, this organism can be readily separated from currently described species of Tissierella. Therefore, based on the phenotypic and phylogenetic distinctiveness of the new isolate, it is proposed that the bacterium be classified as a new species of the genus Tissierella, Tissierella creatinophila sp. nov. The type strain is KRE 4 (= DSM 69113.

Genome Sequence of Creatinine-Fermenting Tissierella creatinophila Strain KRE 4 T (DSM 6911)

“…The strictly anaerobic, Gram-positive, and non-spore-forming Tissierella creatinophila strain KRE 4 T is able to grow on creatinine or on creatinine plus formate as a sole source of carbon, nitrogen, and energy (1). Strain KRE 4 T was shown to degrade creatinine via creatine, sarcosine, and glycine to acetate, monomethylamine, ammonia, and carbon dioxide (1, 2).…”

“…Strain KRE 4 T was shown to degrade creatinine via creatine, sarcosine, and glycine to acetate, monomethylamine, ammonia, and carbon dioxide (1, 2). The growth of this strain with creatinine or creatine as substrates was selenium-dependent and stimulated by formate (1). T. creatinophila strain KRE 4 T (DSM 6911 T ) was isolated from sewage sludge and obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ).…”

“…Analysis of the genome revealed that T. creatinophila strain KRE 4 T harbors two putative creatinine amidohydrolase genes. Furthermore, genes encoding cytosine deaminase, N -carbamoylsarcosine amidase, glycine reductase, as well as sarcosine reductase complex components A, B, and C, 5,10-methylenetetrahydrofolate reductase, methenyltetrahydrofolate cyclohydrolase, formate-tetrahydrofolate ligase, and formate dehydrogenase, which are associated with degradation of creatinine (1, 7), were identified. We also detected a gene encoding a D-hydantoinase-like protein forming a cluster with the gene for the N -carbamoylsarcosine amidase, which might be involved in the degradation of 1-methylhydantion to N -carbamoylsarcosine (7).…”

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.…”

“…On the basis of its phylogenetic, genetic and physiological (Gauglitz, 1988;Hermann et al, 1992); 3, Tissierella praeacuta (Tissier, 1908;Cato et al, 1979;Collins & Shah, 1986;Farrow et al, 1995); 4, Tissierella creatinophila (Harms et al, 1998); 5, C. hastiforme (Holdeman et al, 1977;Cato et al, 1982;Suen et al, 1988);6, C. ultunense (Schnürer et al, 1996). 2, Negative; +, positive; NR, not reported; W, weak.…”