The growth factor and amino acid requirements of species of the genus Leuconostoc, including Leuconostoc paramesenteroides (sp.nov.) and Leuconostoc oenos.
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Amino Acids↗
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A phylogenetic analysis of an atypical leuconostoc: description of Leuconostoc fallax sp. nov.
The 16S rRNA sequence of an unknown leuconostoc originally isolated from sauerkraut was investigated by reverse transcription. A comparison of the sequence with those from other lactic acid bacteria revealed the unknown organism represents a new albeit peripheral line within the genus Leuconostoc sensu stricto. A new species, Leuconostoc fallax, is proposed for this organism. The type strain is DSM 20189.
Base Sequence↗
Taxonomic studies on some leuconostoc-like organisms from fermented sausages: description of a new genus Weissella for the Leuconostoc paramesenteroides group of species.
Taxonomic studies were performed on some unknown Leuconostoc-like organisms from fermented Greek sausage. Comparative 16S rRNA sequence analysis showed the unidentified organisms represent a new line within the Leuconostoc paramesenteroides group of species. On the basis of the results of this and earlier phylogenetic investigations, it is proposed that Leuconostoc paramesenteroides and related species be reclassified in a new genus Weissella. In addition a new species, Weissella hellenica, is proposed for the isolates from fermented sausage.
Base Sequence↗
Cell wall constituents of Leuconostoc citrovorum and Leuconostoc mesenteroides.
The cell wall constituents of Leuconostoc citrovorum 8082, L. mesenteroides 10830a, and L. mesenteroides 11449 have been ascertained. All three strains contained glycerol. Glucose and rhamnose were the major reducing sugar constituents. Alanine, glutamic acid, lysine, glucosamine, and muramic acid were the principal amino acids and amino sugars in all three strains. In addition, strain 10830a contained l-serine as a major cell wall component. Quantitative amino acid analyses indicate that glutamic acid, lysine, glucosamine, muramic acid, and serine may be present in the cell walls in equimolar amounts and that alanine is present in three to four times these quantities. The similarities and differences between the cell wall constituents of the leuconostocs and those of the lactobacilli and streptococci are discussed.
Alanine↗
Occurrence of Leuconostoc mesenteroides and leuconostoc-like organisms in Lagos, Nigeria.
A total of 91 catalase--negative Gram-positive coccal isolates obtained from 245 clinical specimens in Lagos were characterized. Ten (11.0%) of the isolates were vancomycin resistant, they fermented glucose, sucrose, fructose, lactose, mannose, mannitol, ribose, salicin, sorbitol, arabinose and xylose with acid production. One of the isolates produced in addition gas inclusive and ethanol, thus identified as Leuconostoc mesenteroides. The ten vancomycin-resistant Gram-positive coccal organisms (VRGPC) showed variable sensitivity patterns to penicillin, tetracycline, erythromycin, ampicillin, streptomycin, chloramphenicol, cloxacillin and co-trimoxazole. The possible role of Leuconostoc spp. and VRGPC in clinical infections in hospital setting is still to be defined.
Bacterial Typing Techniques↗
Inter-strain relationships among wine leuconostocs and their divergence from other Leuconostoc species, as revealed by low frequency restriction fragment analysis of genomic DNA.
Thirty Leuconostoc oenos strains, representing 28 different isolates, were distributed into 20 genomic groups according to PFGE patterns of restriction digests. The 8 bp-specific enzymes Sfi I, Not I and Asc I cleaved the Leuc. oenos DNA in a mean of 17, 11 and four fragments respectively and Sma I produced more than 50 fragments per genome. The strain differentiating capacity of the four enzymes was similar; only two related genomic groups failed to be distinguished by Asc I or Not I. Genomic relationships between Leuc. oenos strains were quantified by numerical analysis of Not I and Sfi I banding patterns. More than half of the strains, including the starters ML34 and PSU-1, formed a major cluster. The average size of the Leuc. oenos genome was estimated as 1.86 Mb. Although similar values were obtained for the genomes of Leuc. mesenteroides, Leuc. pseudomesenteroides, Leuc. gelidum and Leuc. citreum, a significant divergence between wine and non-wine species was inferred from comparisons of genome cleavage frequencies, determined with five different enzymes.
Base Sequence↗
Primary structure of dextransucrase from Leuconostoc mesenteroides NRRL B-1299. Comparison with dextransucrase from Leuconostoc mesenteroides NRRL B-512F.
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Amino Acid Sequence↗
Development and use of a selective medium for isolation of Leuconostoc spp. from vegetables and dairy products.
A selective medium (LUSM medium) for the isolation of Leuconostoc spp. was developed. This medium contained 1.0% glucose, 1.0% Bacto Peptone (Difco), 0.5% yeast extract (BBL), 0.5% meat extract (Difco), 0.25% gelatin (Difco), 0.5% calcium lactate, 0.05% sorbic acid, 75 ppm of sodium azide (Sigma), 0.25% sodium acetate, 0.1% (vol/vol) Tween 80, 15% tomato juice, 30 micrograms of vancomycin (Sigma) per ml, 0.20 microgram of tetracycline (Serva) per ml, 0.5 mg of cysteine hydrochloride per ml, and 1.5% agar (Difco). LUSM medium was used successfully for isolation and enumeration of Leuconostoc spp. in dairy products and vegetables. Of 116 colony isolates obtained from fresh raw milk, curdled milk, or various vegetables, 115 were identified as members of the genus Leuconostoc. A total of 89 of these isolates were identified to species; 13.5% of the isolates were Leuconostoc cremoris, 7.9% were Leuconostoc mesenteroides subsp. mesenteroides, 11.2% were Leuconostoc mesenteroides subsp. dextranicum, 16.9% were Leuconostoc mesenteroides subsp. paramesenteroides, 10.1% were leuconostoc lactis, and 40.4% were Leuconostoc oenos. When we compared the counts obtained for two Leuconostoc strains, Leuconostoc dextranicum 181 and L. cremoris JLL8, on MRS agar and LUSM medium, we found no significant difference between the values obtained on the two media.
Bacteriological Techniques↗
Bacteriocins produced by Leuconostoc species.
Leuconostoc spp. are lactic acid bacteria that are commonly associated with foods and that are used as starter bacteria in some dairy fermentations. Lactic acid bacteria are inhibitory to other bacteria because of pH, organic acids, hydrogen peroxide, and other chemicals produced during their growth, including bacteriocins. Bacteriocin production by Leuconostoc spp. was first observed in the 1950s, but only since 1984, when antagonistic activity of Leuconostoc spp. was reported, have more extensive studies of bacteriocins produced by Leuconostoc spp. been conducted, including mesentericin Y105, produced by Leuconostoc mesenteroides spp. mesenteroides; leucocin A-UAL 187, produced by Leuconostoc gelidum; carnosin 44A, produced by Leuconostoc carnosum; and leuconocin S, produced by Leuconostoc paramesenteroides. Bacteriocins produced by leuconostocs may or may not be active against other lactic acid bacteria, but all include Listeria in their activity spectra. Mesentericin Y105 is reported to be exclusively active against Listeria spp. The amino acid sequences for leucocin A and mesentericin Y105 have been determined. Despite considerable differences in antibacterial spectra, only two amino acids differ between these bacteriocins. The prevalence of leuconostocs in many adventitious fermentations of food and the use of leuconostocs as starter bacteria in controlled fermentations make the bacteriocins produced by these bacteria of interest as possible food preservatives by addition of the bacteriocin or its producer organism to foods.
Amino Acid Sequence↗
Antibacterial activity of three Leuconostoc strains isolated from vacuum-packaged processed meats.
One hundred and fifty lactic acid bacteria (LAB) isolated from vacuum-packaged processed meats were screened for antagonistic activity against various food spoilage microorganisms and foodborne pathogens. Nineteen strains produced bacteriocins active against closely related LAB and Listeria strains. Leuconostoc carnosum (LA54a and TA26b) and Leuconostoc mesenteroides subspecies dextranicum (TA33a) produced bacteriocins that were susceptible to proteolytic enzymes, but not to catalase, lysozyme or chloroform. They were heat stable up to 100 degrees C for thirty minutes at pH 2 to 7, and exerted a bacteriolytic effect. Bacteriocin production by all Leuconostoc strains was growth associated, occurring at incubation temperatures of 0 degrees C to 30 degrees C and initial medium pH 4.5 to 7.5. Probing of plasmid DNA from the three Leuconostoc strains with an oligonucleotide probe homologous to the nucleotide sequence of leucocin A-UAL 187 indicated plasmid-mediated bacteriocin production. Homology of the three Leuconostoc bacteriocin-coding genes to the amino-terminal end of the leucocin A-UAL 187 gene from Leuconostoc gelidum UAL 187 is therefore suggested. This evidence implies that all three Leuconostoc strains produce type 2, Listeria active bacteriocins.
Bacteria↗
Bacteriocins of leuconostocs isolated from meat.
Several bacteriocin-producing Leuconostoc strains have been isolated from meat and identified as Leuconostoc gelidum UAL 187, Leuconostoc paramesenteroides-La7a, Leuconostoc carnosum-Ta11a and Leuconostoc carnosum-La54a. All strains produce bacteriocins that are active against Listeria monocytogenes and other lactic acid bacteria of concern in meat spoilage. All of the bacteriocins studied are heat stable in acidic environments and are inactivated by a range of proteolytic enzymes but not by catalase or lysozyme. Most are detected early in the growth cycle and are produced at refrigeration temperatures and in a pH range of 4.0-7.0. Leucocin A-UAL187, produced by Leuconostoc gelidium UAL 187, is a small peptide (MW 3930) translated as a 61 amino acid prepeptide consisting of a 24 amino acid leader region and 37 amino acid active bacteriocin that is secreted. A probe designed from a region of the leucocin gene has been used to locate the bacteriocin genes in the other strains (La7a, La54a and Ta11a). Strong hybridization signals were detected from 8.9 MDa, 32 MDa and 8.9 MDa plasmids in strains La7a, La54a and Ta11a, respectively. The bacteriocin structural gene from Leuconostoc carnosum-Ta11a (leucocin B-Ta11a) has been cloned and sequenced and the bacteriocin shows 100% homology to leucocin A-UAL187; however, the prepeptide differs in six residues. The mature extracellular bacteriocin from strain UAL 187 was purified and characterized by precipitation, gel filtration, hydrophobic interaction chromatography followed by RP-HPLC and amino-terminal sequencing, whilst those of the other strains are in the process of being purified and characterized using similar techniques.(ABSTRACT TRUNCATED AT 250 WORDS)
Amino Acid Sequence↗
Proposal to reclassify Leuconostoc oenos as Oenococcus oeni [corrig.] gen. nov., comb. nov..
Wine strains belonging to the genus Leuconostoc were classified as Leuconostoc oenos by Garvie in 1967, and this name was confirmed on the Approved Lists of Bacterial Names in 1980. L. oenos is distinguished from other Leuconostoc spp. by its growth in acidic media, by its requirement for a growth factor in tomato juice, and by a number of carbohydrate fermentation characteristics. In addition, the results of a total soluble cell protein analysis, an electrophoretic analysis of NAD-dependent D-(-)-lactate dehydrogenase, 6-phosphogluconate dehydrogenase, and alcohol dehydrogenase, and an analysis of cross-reactivity with anti-glucose-6-phosphate dehydrogenase and anti-NAD-dependent D-(-)-lactate dehydrogenase performed with other Leuconostoc spp. clearly indicated that L. oenos should be distinguished from the other Leuconostoc species. Phylogenetic studies, in particular 16S and 23S rRNA sequencing studies, have revealed that L. oenos represents a distinct subline that is separate from other Leuconostoc spp. and lactic acid bacteria. In view of the phenotypic and phylogenetic distinctiveness of L. oenos, we propose that this species should be assigned to a new genus as Oenococcus oeni [corrig.] gen. nov., comb. nov. The type strain of O. oeni is NCDO 1674 (= ATCC 23179).
DNA, Bacterial↗
Analysis of the beta' subunit of DNA-dependent RNA polymerase does not support the hypothesis inferred from 16S rRNA analysis that Oenococcus oeni (formerly Leuconostoc oenos) is a tachytelic (fast-evolving) bacterium.
rRNA sequencing has shown that leuconostocs comprise three distinct phylogenetic lineages which have been designated separate genera (viz., the genera Leuconostoc sensu stricto, Oenococcus, and Weissella). In addition, the 16S rRNA line formed by Oenococcus oeni (formerly Leuconostoc oenos) is exceptionally long; this fact, together with variations in the compositions of conserved positions in the 16S rRNA, has led to the hypothesis (D. Yang and C. R. Woese, Syst. Appl. Microbiol. 12:145-149, 1989) that this organism is a fast-evolving bacterium. Previous evidence that the leuconostocs should be divided into three genera and that O. oeni is an example of tachytelic evolution has come solely from rRNA analyses. In this study we seqenced the rpoC gene encoding the beta' subunit of DNA-dependent RNA polymerase of leuconostocs and performed a comparative phylogenetic analysis. The subdivision of the leuconostocs into three distinct lineages was confirmed by the rpoC gene data, but no evidence that indicated that O. oeni is evolving at an extraordinary rate was found. If O. oeni is truly tachytelic, then fast-evolving phenomena would be expected to occur throughout the whole genome, including this independent molecular chronometer.
Amino Acid Sequence↗
Common occurrence of plasmid DNA and vancomycin resistance in Leuconostoc spp.
Resistance to vancomycin permitted detection, in a culture of Streptococcus cremoris 290PC, of a contaminant gram-positive coccus. Morphological and physiological characteristics indicated that this bacterium was a strain of Leuconostoc sp., designated PO184. This strain contained four plasmid species, which were distinct from those harbored by S. cremoris 290PC. Antibiotic disk susceptibility tests indicated that Leuconostoc sp. strain PO184 was also resistant to sulfathiazole and trimethoprim and susceptible to 17 other antimicrobials. The MIC of vancomycin for this strain was greater than 2,000 micrograms/ml, and resistance did not depend on drug inactivation. Leuconostoc sp. strain PO184 produced a substance which was inhibitory to S. cremoris U134, but not to S. lactis ATCC 11454. Five other leuconostocs produced substances with antibacterial activity. Of 18 strains of Leuconostoc sp., 14 were resistant to at least 500 micrograms of vancomycin per ml, including four L. oenos strains which harbored no plasmid DNA in the 1- to 76-megadalton range. Twelve Leuconostoc sp. strains contained at least one plasmid species in this mass range. These findings are discussed from the physiological, taxonomical, and ecological standpoints and with regard to their potential applications.
DNA, Bacterial↗
Differentiation of dextran-producing Leuconostoc strains from fermented rice cake (puto) using pulsed-field gel electrophoresis.
Lactic acid bacteria were isolated from puto, a fermented rice cake consumed as a breakfast and snack food in the Philippines. The microflora was dominated by dextran-producing leuconostocs, and these were differentiated into four groups using pulsed-field gel electrophoresis of restriction enzyme digested chromosomal DNA, in conjunction with taxonomic tests. The four groups corresponded to the species Leuconostoc mesenteroides subsp. mesenteroides, Leuconostoc pseudomesenteroides, Leuconostoc citreum and Leuconostoc fallax. Several strains showed an unusual clumping phenotype, and two of these were capable of inhibiting other strains of lactic acid bacteria.
Bacteriocins↗
Infection due to Leuconostoc species: six cases and review.
Leuconostocs, members of the family Streptococcacae, have only recently been recognized as potential pathogens. We describe six cases of leuconostoc bacteremia and review 11 additional cases of infection reported in the literature. Fifteen patients with bacteremia ranged from neonates to persons aged 78 years. Almost all were hospitalized with significant underlying diseases, had received previous antibiotic therapy, and had undergone procedures that interrupted the normal integumentary defense. Leuconostoc bacteremia was associated with fever, leukocytosis, and gastrointestinal complaints. Eight of 15 patients had polymicrobial bacteremia, seven of these eight with staphylococcal species. Clinical isolates of Leuconostoc were frequently misidentified, usually as viridans streptococci. All clinical isolates identified to date--and most agricultural isolates--demonstrate a high level of resistance to vancomycin. Successful regimens for treatment of Leuconostoc include high-dose penicillin, clindamycin, and where appropriate, removal of infected intravascular catheters. Susceptibility testing of all gram-positive bacteria isolated from normally sterile body sites is recommended.
Adolescent↗
The putative immunity protein of the gram-positive bacteria Leuconostoc mesenteroides is preferentially located in the cytoplasm compartment.
Immunity proteins are though to protect bacteriocin-producing bacterial strains against the bactericidal effects of their own bacteriocin. The immunity protein which protects the lactic acid bacterium Leuconostoc mesenteroides against mesentericin Y105(37) bacteriocin was detected and localized by immunofluorescence and electron microscopy, using antibodies directed against the C-terminal end of the predicted immunity protein. The antibodies recognized the immunity proteins of various strains of Leuconostoc, including Leuconostoc mesenteroides and Leuconostoc gelidum. This study demonstrated that immunity proteins produced by Leuconostoc mesenteroides accumulated in the cytoplasmic compartment of the bacteria. This is in contrast with other known immunity proteins, such as the colicin immunity proteins, which are integral membrane proteins possessing three to four transmembrane domains.
Amino Acid Sequence↗
An automated RNA extraction procedure and application for 16S rRNA sequencing of Leuconostoc amelobiosum.
The determination of 16S ribosomal ribonucleic acid (16S rRNA) primary structures by in vitro reverse transcription requires the extraction of rRNA in pure form. Although a number of high reliable techniques have been developed for the purpose most are fairly complex, involving numerous steps and the wasting of large volumes. It describes here a RNA extraction and purification method, suitable for automatic extractors, which consistently yields reasonable amounts of pure total RNA from prokaryotes, free of DNA and RNases. The rRNA from the type strain Leuconostoc amelobiosum was isolated using this procedure. Its 16S rRNA sequence was determined and a comparative analysis with those from all currently described leuconostocs, including several atypical lactobacilli, revealed very high sequence homology with Leuconostoc citreum confirming Leuconostoc amelobiosum is phylogenetically a member of the genus Leuconostoc sensu stricto.
Base Sequence↗