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

Abstract

Antisera prepared against the glyceraldehyde-3-phosphate dehydrogenases of Streptococcus faecalis, Pediococcus damnosus, and Lactobacillus acidophilus were used to measure relationships among the lactic acid bacteria by immunochemical techniques. Our results confirmed the results of earlier phylogenetic studies carried out with anti-fructose diphosphate aldolase sera. 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. The aldolase-based cluster was comprised of the following genera: Acholeplasma, Aerococcus, Brochothrix, Eubacterium, Lactobacillus, Pediococcus, Propionibacterium, Arachnia, and Streptococcus. With their lactate dehydrogenase studies, Gasser and Gasser (9) had previously established that certain species of Lactobacillus and Leuconostoc are related to one another. These relationships were extended and refined by subsequent work in which glucose-6-phosphate dehydrogenase was used as an evolutionary marker (10, 12).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 (16-18). Therefore, it was essential that the conclusions drawn from the aldolase studies be confirmed and extended by a comparative survey with a second enzyme. To this end, the glycolytic enzyme glyceraldehyde-3-phosphate (GA3P) dehydrogenase was selected as a reference protein for the study described in this paper. The selection of this enzyme had the additional virtue of permitting the heterofermentative lactic acid bacteria to be included in this study. Antisera against the purified GA3P dehydrogenases of Streptococcus faecalis ATCC 27792, Lactobacillus acidophilus ATCC 4356= (T = type strain), and Pediococcus damnosus NIRD 559 were used to test and verify the results of the aldolase study. In this report we describe the experiments carried out with the anti-GA3P dehydrogenase sera. MATERIALS AND METHODSMicroorganisms and cultivation. The strains of bacteria used in this study are listed in Table 1. The app...

Polyphasic taxonomy, a consensus approach to bacterial systematics.

“…This difference could not be demonstrated by polyacrylamide disk gel electrophoresis (162), and therefore the authors advised the use of the large difference in the moles percent GϩC of the DNA (35 to 37 mol% for L. jensenii versus 49 to 51 mol% for L. delbrueckii) to differentiate the species. Quantitative immunological techniques have been used with LDH as an evolutionary marker besides numerous other enzymes like fructose-1,6-bisphosphate aldolase (188,190,191), malic enzymes (192,193), and glyceraldehyde-3-phosphate dehydrogenase (189). The data were used to create dendrograms and (three-dimensional) phylogenetic maps.…”

Lactococcus lactis glyceraldehyde-3-phosphate dehydrogenase gene, gap: further evidence for strongly biased codon usage in glycolytic pathway genes

“…These four glycolytic genes were found to exhibit a strong bias in their codon usage (Cancilla e t al., 1995 ation of glyceraldehyde 3-phosphate to produce 1,3-bisphosphoglycerate, using NAD' as a coenzyme (Harris & Waters, 1976). This enzyme, in conjunction with 3-phosphoglycerate kinase (EC 2.7.2.3), forms a central link between ATP production and NAD' reduction, and hyde-3-phosphate dehydrogenases of Streptococct/sfaecal, Pediococcxr damnosm and Lactobacilhs acidophilzls were used to demonstrate the relationships between five genera of Gram-positive lactic acid bacteria (London & Chace, 1983). therefore has a key role in glycolysis.…”

Restriction Endonuclease Patterns and Multivariate Analysis as a Classification Tool for Lactobacillus spp.

“…Sequencing of rRNA is technically difficult and labor intensive. The accuracy of using enzymes as genetic markers (15) can be questioned as long as the majority of the information in the DNA remains unknown. And DNA-DNA homology is, unfortunately, not suitable outside the constraints of highly related species.…”