Identification of "Staphylococcus staphylolyticus" NRRL B-2628 as a Biovar of Staphylococcus simulans

Sloan, Gary L.; Robinson, John M.; Kloos, Wesley E.

doi:10.1099/00207713-32-2-170

Cited by 43 publications

(30 citation statements)

References 24 publications

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“…d Strain NRRL B-2628 (DSM 20723), originally described as "S. staphylolyticus," was classified as a biovar of S. simulans (34 Comparison of RIDOM and NCBI database queries. A total of 55 clinical staphylococcal isolates were analyzed by 16S rDNA sequencing.…”

Section: Resultsmentioning

confidence: 99%

Development and Evaluation of a Quality-Controlled Ribosomal Sequence Database for 16S Ribosomal DNA-Based Identification of Staphylococcus Species

Becker¹,

et al. 2004

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To establish an improved ribosomal gene sequence database as part of the Ribosomal Differentiation of Microorganisms (RIDOM) project and to overcome the drawbacks of phenotypic identification systems and publicly accessible sequence databases, both strands of the 5 end of the 16S ribosomal DNA (rDNA) of 81 type and reference strains comprising all validly described staphylococcal (sub)species were sequenced. Assuming a normal distribution for pairwise distances of all unique staphylococcal sequences and choosing a reporting criterion of >98.7% similarity for a "distinct species," a statistical error probability of 1.0% was calculated. To evaluate this database, a 16S rDNA fragment (corresponding to Escherichia coli positions 54 to 510) of 55 clinical Staphylococcus isolates (including those of the small-colony variant phenotype) were sequenced and analyzed by the RIDOM approach. Of these isolates, 54 (98.2%) had a similarity score above the proposed threshold using RIDOM; 48 (87.3%) of the sequences gave a perfect match, whereas 83.6% were found by searching National Center for Biotechnology Information (NCBI) database entries. In contrast to RIDOM, which showed four ambiguities at the species level (mainly concerning Staphylococcus intermedius versus Staphylococcus delphini), the NCBI database search yielded 18 taxon-related ambiguities and showed numerous matches exhibiting redundant or unspecified entries. Comparing molecular results with those of biochemical procedures, ID 32 Staph (bioMérieux, Marcy I'Etoile, France) and VITEK 2 (bioMérieux) failed to identify 13 (23.6%) and 19 (34.5%) isolates, respectively, due to incorrect identification and/or categorization below acceptable values. In contrast to phenotypic methods and the NCBI database, the novel high-quality RIDOM sequence database provides excellent identification of staphylococci, including rarely isolated species and phenotypic variants.The emerging role of coagulase-negative staphylococci (CoNS), along with Staphylococcus aureus, in connection with the expanding number of staphylococcal (sub)species described necessitates their comprehensive and accurate identification. A variety of methods have been proposed for the identification of staphylococcal species, including conventional identification schemes based on publications of Kloos and Schleifer (18) and commercial identification tests. In general, methods based on phenotypic characteristics are hampered by the fact that they are dependent on the expression of metabolic activities and/or morphological features. Furthermore, commercial systems often offer two or more suggestions for identification with a comparable safety level.Nowadays, with the advent of molecular biology-based techniques, investigations based on comparative DNA sequence analysis of the genes of conserved macromolecules have become commonplace in microbiology as a tool for classification of microbial organisms. The most useful and extensively investigated taxonomic marker molecules are the larger rRNAs and their corresponding ...

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Section: Resultsmentioning

confidence: 99%

Development and Evaluation of a Quality-Controlled Ribosomal Sequence Database for 16S Ribosomal DNA-Based Identification of Staphylococcus Species

Becker¹,

et al. 2004

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“…simulans biovar staphy/olyticus ATCCl362 (NRRL B-2628) formerly described as 'S. staphylolyticus' (Sloan et a/., 1982), is the lysostaphin-and cysteine-protease-producing wild-type strain. The lysostaphin-negative strain, S. simulans biovar staphylolyticus (ApACKl), was made by curing S. sirnutans biovar staphyldyficus ATCC1362 of plasmid pACK1, which contains the lysostaphin gene (lss), using ethidium bromide (Gotz, 1983).…”

Section: Bacterial Strains Expression Vectors and Growth Conditionsmentioning

confidence: 99%

“…Lysostaphin is an extracellular enzyme produced by a single known staphylococcal strain, formerly designated 'Sfaphylococcus sfaphylolyticus' and now designated as Sfaphylococcus simulans biovar sfaphylolyticus ATCCI 362 (NRRL 8-2628) (Schindler and Schuhardt, 1964;Sloan et a/., 1982). As a cell-wall lytic enzyme, lysostaphin is indispensable for staphylococcal genetic studies and is used for DNA isolation, formation of protoplasts, and differentiation of staphylococcal strains.…”

Section: Introductionmentioning

confidence: 99%

Studies on prolysostaphin processing and characterization of the lysostaphin immunity factor (Lif) of Staphylococcus simulans biovar staphylolyticus

Thumm¹,

Götz²

1997

Molecular Microbiology

155

162

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SummaryLysostaphin is an extracellular glycylglycine endopeptidase produced by Staphylococcus simulans biovar staphyIolyticus ATCC1362 that lyses staphylococcal cells by hydrolysing the polyglycine interpeptide bridges of the peptidoglycan. Renewed analysis of the sequence of the lysostaphin gene (Iss), and the sequencing of the amino-terminus of purified prolysostaphin and of mature lysostaphin revealed that lysostaphin is organized as a preproprotein of 493 amino acids (aa), with a signal peptide consisting of 36aa, a propeptide of 21 1 aa from which 195 aa are organized in 15 tandem repeats of 13 aa length, and a mature protein of 246aa. Prolysostaphin is processed in the culture supernatant of S. simulans biovar StaphyIoIyficus by an extracellular cysteine protease. Although prolysostaphin was staphylolytically active, the mature lysostaphin was about 4.5-fold more active. The controlled expression in Staphylococcus carnosus of lss and lss with deletions in the prepropeptide region indicated that the tandem repeats of the propeptide are not necessary for protein export or activation of Lss, but keep Lss in a less active state. lntracellularly expressed pro-and mature lysostaphin exert staphylolytic activity in cell-free extracts, but do not affect growth of the corresponding clones. We characterized a lysostaphin immunity factor gene ( / i f ) which is located in the opposite direction to lss. The expression of /if in s. carnosus led to an increase in the serine/glycine ratio of the interpeptide bridges of peptidoglycan from 2 to 35%, suggesting that lysostaphin immunity depends on serine incorporation into the interpeptide bridge. If, in addition to /if, Iss is coexpressed the serinelglycine ratio is further increased to 58%, suggesting that Lss selects for optimal serine incorporation. Lif shows similarity to FemA and FemB

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“…rodentiurn, and designate the type strains of these taxa. The nature of two surprising phenotypic properties, common methicillin resistance and staphylolytic activity, were explored in some detail, because these properties indicate that S. sciuri is a natural reservoir of important methicillin resistance and staphylolytic enzyme genes and their products that are novel to certain other Staphylococcus species (5,21,39,43,53,54). …”

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Ribotype Delineation and Description of Staphylococcus sciuri Subspecies and Their Potential as Reservoirs of Methicillin Resistance and Staphylolytic Enzyme Genes

Kloos

Ballard

Webster

et al. 1997

International Journal of Systematic Bacteriology

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Three subspecies of Staphylococcus sciuri, S. sciuri subsp. sciuri Moos, Schleifer, and Smith 1976, 23& emend. Moos et al. 1996, S. sciuri subsp. curnaticus subsp. nov., and S. sciuri subsp. rodentium subsp. nov., are described on the basis of their ribotype patterns, DNA-DNA liquid hybridization data, and phenotypic characteristics. Normalized ribotyping subdivided the S. sciuri patterns into three blocks of patterns, each corresponding to a subspecies. Each subspecies formed a separate, well-defined DNA similarity group when DNA-DNA hybridizations were conducted under stringent (7OOC) reassociation conditions. S. sciuri subsp. sciuri could be distinguished from the other subspecies on the basis of its ability to produce acid from D-cellobiose, alkaline phosphatase activity, and inability to produce either clumping factor or protein A. S. sciuri subsp. curnaticus could be distinguished by its ability to produce acid aerobically from D-xylose and maltose, inability to produce acid from D-melezitose, and smaller colony size on P agar. S. sciuri subsp. rodentium could be distinguished by its positive reaction in the latex agglutination test for clumping factor and/or protein A and generally higher frequencies and levels of oxacillin and methicillin resistance. All 40 strains of S. sciuri tested (including representatives of all three subspecies) hybridized with the mecA gene probe. All strains of S. sciuri subsp. sciuri, 79% of the strains of S. sciuri subsp. carnuticus and 89% of the strains of S. sciuri subsp. rodentium exhibited extracellular, staphylolytic enzyme activity. This activity was associated with an enzyme(s) that immunoblotted with a lysostaphin-specific monoclonal antibody; however, only three strains hybridized with a lysostaphin (end) gene probe. The type strain of S. sciuri subsp. curnaticus is DD 791 (= ATCC 700058), and the type strain of S. sciuri subsp. rodentium is DD 4761 (= ATCC 700061).Staphylococcus sciuri is generally considered one of the most primitive Staphylococcus species; it is widely distributed in nature, is capable of growth on inorganic nitrogen salts as the sole source of nitrogen, and exhibits a wide range of biochemical activities (31-33). In the past 20 years since its original description (33), numerous laboratories have reported frequent isolation of this species from foods, farm animals, rodents, marsupials, marine mammals, and birds and occasional isolation from humans and their pets (1,2, 17,28,31,35,48,52,57,58).Most resident populations of S. sciuri are members of the normal cutaneous microflora of lower mammals. This species is rarely associated with infections. A preliminary investigation of a large collection of S. sciuri strains showed that a ribotyping method based on an analysis of genomic EcoRI fragments containing portions of the rRNA operons could be used to distinguish two major subgroups and additional strain variation within the species (10).In this study, we describe three subpopulations of S. sciuri which can be distinguished on the basis of their rib...

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Identification of "Staphylococcus staphylolyticus" NRRL B-2628 as a Biovar of Staphylococcus simulans

Cited by 43 publications

References 24 publications

Development and Evaluation of a Quality-Controlled Ribosomal Sequence Database for 16S Ribosomal DNA-Based Identification of Staphylococcus Species

Development and Evaluation of a Quality-Controlled Ribosomal Sequence Database for 16S Ribosomal DNA-Based Identification of Staphylococcus Species

Studies on prolysostaphin processing and characterization of the lysostaphin immunity factor (Lif) of Staphylococcus simulans biovar staphylolyticus

Ribotype Delineation and Description of Staphylococcus sciuri Subspecies and Their Potential as Reservoirs of Methicillin Resistance and Staphylolytic Enzyme Genes

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