Vasana Tolieng

Terrilactibacillus laevilacticus gen. nov., sp. nov., isolated from soil.

A Gram-stain-positive, catalase-positive, facultatively anaerobic, spore-forming, rod-shaped bacterium, strain NK26-11T, was isolated from soil in Thailand. This strain produced d-lactic acid from glucose homofermentatively, and grew at 20-45 °C and pH 5-8.5. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The major respiratory quinone was menaquinone 7 (MK-7), the DNA G+C content was 42.6 mol%, and the major cellular fatty acids were anteiso-C15 : 0 and anteiso-C17 : 0. On the basis of 16S rRNA gene sequences analysis, strain NK26-11T was closely related to Bacillus solimangrovi JCM 18994T (93.89 % 16S rRNA gene sequence similarity), Pullulanibacillus naganoensis LMG 12887T (93.32 %), Sporolactobacillus inulinus NRIC 1133T (92.99 %), Tuberibacillus calidus JCM 13397T (92.98 %) and Thalassobacillus devorans DSM 16966T ( < 90.93 %). Strain NK26-11T could be clearly distinguished from the closely related genera based on phenotypic characteristics and DNA G+C content, and thus represents a novel species of a new genus between the Bacillus and Sporolactobacillus cluster, for which the name Terrilactibacillus laevilacticus gen. nov., sp. nov. is proposed. The type strain of the type species is NK26-11T ( = LMG 27803T = TISTR 2241T = PCU 335T).

Characterization of D -lactic acid, spore-forming bacteria and Terrilactibacillus laevilacticus SK5-6 as potential industrial strains

In this study, we screened and isolated D-lactic acid-producing bacteria from soil and tree barks collected in Thailand. Among the isolates obtained, Terrilactibacillus laevilacticus SK5-6 exhibited good D-lactate production in the primary screening fermentation (99.27 g/L final lactate titer with 0.90 g/g yield, 1.38 g/L⋅h, and 99.00% D-enantiomer equivalent). Terrilactibacillus laevilacticus SK5-6 is a Gram-positive, endospore-forming, homofermentative D-lactate producer that can ferment a wide range of sugars to produce D-lactate. Unlike the typical D-lactate producers, such as catalase-negative Sporolactobacillus sp., T. laevilacticus SK5-6 possesses catalase activity; therefore, a two-phase fermentation was employed for D-lactate production. During an aerobic preculture stage, a high-density cell mass was rapidly obtained due to aerobic respiration. When transferred to the fermentation stage at the correct physiological stage (inoculum age) and proper concentration of cell mass (inoculum size), T. laevilacticus rapidly converted glucose into D-lactate under anaerobic conditions, resulting in a high final lactate titer (102.22 g/L), high yield (0.84 g/g), and high productivity (2.13 g/L⋅h). When the process conditions were shifted from an aerobic to an anaerobic environment, unlike other lactate-producing bacteria, the mixed acid fermentation route was not activated in the culture of T. laevilacticus SK5-6 during the fermentation stage when some trace oxygen still remained. Our study demonstrates the excellent characteristics of this isolate for D-lactate production; in particular, a high product yield was obtained without byproduct formation. Based on these key characteristics of T. laevilacticus SK5-6, we suggest that this isolate is a novel D-lactate producer for use in industrial fermentation.

Sporolactobacillus shoreicorticis sp.nov., a lactic acid-producing bacterium isolated from tree bark

A Gram-stain-positive, lactic acid-producing bacterium designed strain MK21-7T, was isolated from tree bark collected from the north east of Thailand. This strain was a facultatively anaerobic spore-forming rod that was catalase-negative. It contained meso-diaminopimelic acid in the cell wall peptidoglycan and had seven isoprene units (MK-7) as the predominant menaquinone. Major fatty acids of MK21-7T were anteiso-C17 : 0, iso-C16 : 0, anteiso-C15 : 0 and C18 : 1ω9c. Polar lipids were phosphatidglycerol, diphosphatidylglycerol, an unknown phospholipid, three unknown glycolipids and an unknown lipid. The results of 16S rRNA gene sequence analysis indicated that it represented a member of the genus Sporolactobacillus. MK21-7T showed the highest 16S rRNA gene sequence similarity to Sporolactobacillus terrae NBRC 101527T with 98.4 % similarity and exhibited 97.6 % similarity with Sporolactobacillus kofuensis NRIC 0334T, 97.5 % with Sporolactobacillus laevolacticus NRIC 0361T, 97.3 % with Sporolactobacillus nakayamaesubsp.nakayamae NRIC 0347T and 97.1 % with Sporolactobacillus nakayamaesubsp.racemicus NBRC 101524T. Analysis of the phylogenetic relationship based on 16S rRNA and gyrB gene sequencing revealed that the position of MK21-7T was clearly separated from all related species of the genus Sporolactobacillus. It had low DNA-DNA relatedness (22.8-57.2 %) with S. terrae NBRC 101527T and related type strains. The DNA G+C content was 43.1 mol%. On the basis of the results of the phenotypic, genotypic and chemotaxonomic studies, MK21-7T should be classified as representing a novel species of the genus Sporolactobacillus for which the name Sporolactobacillus shoreicorticis sp. nov. is proposed. The type strain is MK21-7T (=NBRC 111517T=LMG 29111T=TISTR 2466T).