Genotypic analyses of lactobacilli with a range of tannase activities isolated from human feces and fermented foods.
A total of 77 tannase producing lactobacilli strains isolated from human feces or fermented foods were examined for their genotypic profiles and intensities of tannase production. With a PCR-based assay targeting recA gene, all strains except one isolate were assigned to either Lactobacillus plantarum, L. paraplantarum, or L. pentosus whereas a 16/23S rDNA targeted PCR-based assay identified all except 6 isolates (inclusive of the above one isolate) as one of the closely related species. Subsequent DNA/DNA hybridization assays revealed that these 6 exceptional isolates showed low homology (between 1.2% and 55.8% relative DNA binding) against type strains of the three species. Supplemental carbohydrate fermentation profiles on the 6 isolates indicated that two of them were identified as L. acidophilus, one as Pediococcus acidilactici, one as P. pentosaceus, and two remained unidentifiable. The evidence suggests that the 16/23S rDNA targeted PCR assay can be used as a reliable identification tool for the closely related lactobacilli, and that the tannase gene is widely distributed within members of the Lactobacillaceae family. Meanwhile, a randomly amplified polymorphism DNA (RAPD) analysis revealed that all except 8 isolates were well allocated in 4 major RAPD clusters, though not species specific, consisting of two L. plantarum predominant clusters, one L. paraplantarum predominant, and one L. pentosus predominant. The RAPD patterns of the 8 non-clustered isolates, which consisted of the 6 unidentifiable isolates and 2 isolates identified as L. pentosus, were <40% similarity to those belonging to the 4 clusters. A quantitative assay of the tannase activities showed that there was a marked variation in the activities among the strains, which did not correlate with either species identification or clustering by RAPD.
Nishitani
Yosuke
Y
Department of Bioscience, Graduate School of Science and Technology, Kobe University, Kobe, Japan.
Sasaki
Eiki
E
Fujisawa
Tomohiko
T
Osawa
Ro
R
D002241:Carbohydrates / Q000032:analysis; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D016000:Cluster Analysis; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D005243:Feces / Q000382:microbiology; D005285:Fermentation; D005516:Food Microbiology; D006801:Humans; D007778:Lactobacillus / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D019105:Random Amplified Polymorphic DNA Technique; D011938:Rec A Recombinases / Q000737:chemistry / Q000235:genetics; D014675:Vegetables / Q000382:microbiology
D002241:Carbohydrates; D004269:DNA, Bacterial; D021903:DNA, Ribosomal Spacer; D011938:Rec A Recombinases; D002265:Carboxylic Ester Hydrolases; C022711:tannase