Aldolase of Lactic Acid Bacteria: Immunological Relationships Among Aldolases of Streptococci and Gram-Positive Nonsporeforming Anaerobes

Abstract

Antiserum prepared against a Streptococcus faecalis fructose diphosphate aldolase (EC 4.1.2.13) was used to measure the extent of immunological homology between the reference enzyme and aldolases of various streptococci and gram-positive nonsporeforming anaerobic bacteria. The majority of streptococci surveyed were isolated from the oral cavity or blood samples. Strains within the species Streptococcus mutans Clarke and Streptococcus mitis (mitior) possessed aldolases that exhibited marked antigenic heterogeneity and thereby contrasted sharply with previously studied species of Streptococcus (London and Kline, 1973). Varying degrees of cross-reactivity were observed between the aldolases of certain species of Eubacterium, Butyribacteriurn, and Propionibacterium and the anti-streptococcal aldolase serum. Evidence is also presented confirming an earlier report by Neimark (1974) that aldolases of Acholeplusrna species react with the anti-S. faecalis aldolase serum.In a previous study, the fructose diphosphate (FDP) aldolase (EC 4.1.2.13) of Streptococcus faecalis ATCC 27792 (formerly strain MR) was used as a reference point to establish evolutionary relationships among various homofermentative lactic acid bacteria by determining the degree of immunological reactivity between their aldolases and a specific anti-S. faecalis aldolase serum (18). Despite significant differences in the quaternary arrangement of the subunits of this enzyme (17), their primary structure was sufficiently conserved to permit a comparison of their antigenic determinants. A preliminary phylogenetic map of streptococci, lactobacilli, and pediococci was constructed from both qualitative and quantitative immunological data (18).In the present study, the streptococcal branch of the phylogenetic map is confirmed and extended with new immunological data for aldolases of previously untested strains. Also, immunological heterogeneity has been detected among the aldolases from strains of streptococci within the species Streptococcus mutans and Streptococcus mitis (mitior). The data indicate that neither species is homogeneous. A s_trong reaction between the aldolase of Butyribacterium rettgeri and anti-S. faecalis aldolase serum prompted the survey reported here for other immunologically related aldolases among grampositive nonsporeforming anaerobic bacteria. The survey includes organisms belonging to the genera Eu b ac terium, Propioni bact erium , and Arac hnia. MATERIALS AND METHODSOrganisms. The strain designations of the organisms are listed in Table 1 Maintenance and cultivation of t h e microorganisms. The streptococci were all maintained on N.I.H. thioglycolate broth (Difco Laboratories, Detroit, Mich.). For enzymological and immunological studi'es, 2 liters of complex streptococcal medium (16) was inoculated with 10 ml of a 12-h-old culture of the appropriate organism and incubated at 37 C for 18 h. Cells were harvested by centrifugation and washed twice with cold 0.05 M potassium phosphate buffer, pH 7.0. Washed cell pellets were stored at -...

Purification, Properties and Immunological Relationship of l(+)‐Lactate Dehydrogenase from Lactobacillus casei

“…This narrow specificity of the intrageneric reaction displayed by L. casei anti-lactate-dehydrogenase is also in opposition to the extremely broad intergeneric reactions found for antisera to streptococcal fructose 1,6-bisphosphate aldolase [16,53,54], antisera to lactate dehydrogenase and L( +)-lactate dehydrogenase from several lactobacilli [18] and for anti-D( --)-lactatedehydrogenase from several leuconostocs [55]. The close interrelationship between the five strains of streptobacteria reported here indicate that the type strain of the three subspecies of L. casei form a group of identical specificity.…”

Proposal to Elevate the Subspecies of Streptococcus mutans to Species Status, Based on Their Molecular Composition

“…However, new dicate a degree of molecular disparity not ex-information on the evolutionary distance bepected within a single species. Indeed, every tween members of the different genospecies (26) molecule or subcellular component isolated and the discovery of a fifth genospecies (14) from S . mutuns and studied comparatively has make it increasingly difficult to consider all shown differences that correlate with the DNA these streptococci as "Streptococcus mutans .…”

Relationships Among Lactic Acid Bacteria Demonstrated with Glyceraldehyde-3-Phosphate Dehydrogenase as an Evolutionary Probe

“…Whereas evidence for common ancestry of lactobacilli and leuconstocs was sought and demonstrated with antisera prepared against six distinct enzyme preparations (9, 10, 12), evidence for intergeneric relatedness among other lactic acid bacteria rested solely on experiments with two FDP aldolases (16)(17)(18). Therefore, it was essential that the conclusions drawn from the aldolase studies be confirmed and extended by a comparative survey with a second enzyme.…”

“…We present new data in the form of dendrograms, which, for the first time, include heterofermentative members of the lactic acid bacteria. Previously acquired quantitative data are integrated with our new data to produce a three-dimensional phylogenetic map which shows the relationships of five genera of gram-positive, asporogenous bacteria.In a series of studies (15,17,18), antisera prepared against the fructose diphosphate (FDP) aldolases of Streptococcus faecalis and Pediococcus damnosus (synonym, Pediococcus cerevisiae) were used to detect structural homologies between these two reference proteins and isofunctional enzymes from representatives of eight genera of gram-positive, nonsporeforming bacteria. Quantitative immunological techniques were used to measure the degree of immunological homology between the reference enzymes and their isologous counterparts, and a preliminary phylogenetic map of the group was generated from these data.…”

“…In a series of studies (15,17,18), antisera prepared against the fructose diphosphate (FDP) aldolases of Streptococcus faecalis and Pediococcus damnosus (synonym, Pediococcus cerevisiae) were used to detect structural homologies between these two reference proteins and isofunctional enzymes from representatives of eight genera of gram-positive, nonsporeforming bacteria. Quantitative immunological techniques were used to measure the degree of immunological homology between the reference enzymes and their isologous counterparts, and a preliminary phylogenetic map of the group was generated from these data.…”