Abstract
Two Gram-stain-positive bacterial strains, designated 213-9(3)T and 30-1(2)T, were isolated from traditional Chinese pickle, and were characterized using a polyphasic taxonomic approach. Results of 16S rRNA gene sequence analysis indicated that strain 213-9(3)T was most closely related to Levilactobacillus paucivorans TMW 1.1424T, Levilactobacillus huananensis 151-2BT and Levilactobacillus lindianensis 220-4T, having 99.7-99.9 % 16S rRNA gene sequence similarities; strain 30-1(2)T was most closely related to Lapidilactobacillus achengensis 247-4T, with 99.4 % 16S rRNA gene sequence similarity. Strain 213-9(3)T shared the highest pheS (93.9 %), rpoA (99.3 %) and concatenated pheS and rpoA (97.5 %) sequence similarities to L. paucivorans TMW 1.1424T. Strain 30-1(2)T had the highest pheS (82.4 %), rpoA (95.5 %) and concatenated pheS and rpoA (91.2 %) sequence similarities to L. achengensis 247-4T. The phylogenetic relationships based on concatenated pheS and rpoA sequences and whole genome sequences were identical to those based on 16S rRNA gene sequences. Strain 213-9(3)T exhibited the highest average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values (92.7 and 48.8 %, respectively) to L. paucivorans DSM 22467T. Strain 30-1(2)T had the highest ANI (84.4 %) and dDDH (32.8 %) values with L. achengensis 247-4T. Acid production from d-galactose, d-glucose, d-mannose, N-acetyl-β-glucosaminidase, arbutin, salicin, cellobiose, maltose, gentiobiose, d-tagatose and gluconate, hydrolysis of aesculin, and activity of cystine arylamidase could differentiate strain 213-9(3)T from L. paucivorans DSM 22467T. Acid production from l-arabinose, d-ribose, d-xylose and d-galactose, and activity of alkaline phosphatase, esterase (C4), α-mannosidase and α-fucosidase could differentiate strain 30-1(2)T from L. achengensis 247-4T. Based upon the data obtained in the present study, two novel species, Levilactobacillus humaensis sp. nov. and Lapidilactobacillus luobeiensis sp. nov., are proposed and the type strains are 213-9(3)T (=CCM 9241T=CCTCC AB 2022115T=JCM 35554T) and 30-1(2)T (=CCM 9240T=CCTCC AB 2022114T=JCM 35553T), respectively.
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