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Ginseng leaves are known to contain high concentrations of bioactive compounds, such as ginsenosides, and have potential as a treatment for various conditions, including fungal infections, cancer, obesity, oxidative stress, and age-related diseases. This study assessed the impact of ginseng leaf extract (GLE) on mast cell-mediated allergic inflammation and atopic dermatitis (AD) in DNCB-treated mice. GLE reduced skin thickness and lymph node nodules and suppressed the expression and secretion of histamine and pro-inflammatory cytokines. It also significantly lowered the production of inflammatory response mediators including ROS, leukotriene C4 (LTC4), prostaglandin E2 (PGE2), cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS). GLE inhibited the phosphorylation of MAPKs (ERK, P38, JNK) and the activation of NF-κB, which are both linked to inflammatory cytokine expression. We demonstrated that GLE's inhibitory effect on mast cell-mediated allergic inflammation is due to the blockade of the NF-κB and inflammasome pathways. Our findings suggest that GLE can be an effective therapeutic agent for mast-cell mediated and allergic inflammatory conditions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Oh, Yoon, Joo, Im and Chun.)

A Gram-staining-positive, motile, aerobic and rod-shaped bacterium, designated strain MA9 T was isolated from wetland soil of ecology park, in Seoul, Republic of Korea. This bacterium was characterized to determine its taxonomic position by using the polyphasic approach. Strain MA9 T grew at 10-37 °C and at pH 6.0-9.5 on TSB. Menaquinone MK-7 was the predominant respiratory quinone and iso-C 15 : 0 , iso-C 16 : 0 and C 16 : 1  ω7 c alcohol were the major fatty acids. The main polar lipids were phosphatidylethanolamine (PE), phosphatidylserine (PS), diphosphatidylglycerol (DPG) and phosphatidylglycerol (PG). The peptidoglycan type of the cell wall was A4α l-Lys-d-Glu. Based on 16S rRNA gene sequencing, strain MA9 T clustered with species of the genus Solibacillus and appeared closely related to S. silvestris DSM 12223 T (97.8 % sequence similarity), S. cecembensis DSM 21993 T (97.6 %), S. isronensis DSM 21046 T (97.6 %) and S. kalamii DSM 101595 T (96.6 %). The G+C content of the genomic DNA was 37.0 mol%. Digital DNA-DNA hybridization between strain MA9 T and type strains of S. silvestris , S. isronensis , S. cecembensis and S. kalamii resulted in values below 70 %. Strain MA9 T could be differentiated genotypically and phenotypically from the recognized species of the genus Solibacillus . The isolate therefore represents a novel species, for which the name Solibacillus palustris sp. nov. is proposed, with the type strain MA9 T (=KACC 22212 T = LMG 32188 T ).

Two novel bacterial strains, designated as SM33 T and NSE70-1 T , were isolated from wet soil in South Korea. To get the taxonomic positions, the strains were characterized. The genomic information (both 16S rRNA gene and draft genome sequence analysis) show that both novel isolates (SM33 T and NSE70-1 T ) belong to the genus Sphingomonas. SM33 T share the highest 16s rRNA gene similarity (98.2%) with Sphingomonas sediminicola Dae20 T . In addition, NSE70-1 T show 96.4% 16s rRNA gene similarity with Sphingomonas flava THG-MM5 T . The draft genome of strains SM33 T and NSE70-1 T consist of a circular chromosome of 3,033,485 and 2,778,408 base pairs with DNA G+C content of 63.9, and 62.5%, respectively. Strains SM33 T and NSE70-1 T possessed the ubiquinone Q-10 as the major quinone, and a fatty acid profile with C 16:0 , C 18:1 2-OH , C 16:1 ω7c/C 16:1 ω6c (summed feature 3) and C 18:1 ω7c/C 18:1 ω6c (summed feature 8) as major fatty acids. The major polar lipids of SM33 T and NSE70-1 T were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid and phosphatidylcholine, respectively. Moreover, genomic, physiological, and biochemical results allowed the phenotypic and genotypic differentiation of strains SM33 T and NSE70-1 T from their closest and other species of the genus Sphingomonas with validly published names. Therefore, the SM33 T and NSE70-1 T represent novel species of the genus Sphingomonas, for which the name Sphingomonas telluris sp. nov. (type strain SM33 T  = KACC 22222 T  = LMG 32193 T ), and Sphingomonas caseinilyticus (type strain NSE70-1 T  = KACC 22411 T  = LMG 32495 T )., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

A novel yellow-pigmented catalase- and oxidase-positive bacterial strain (designated NA20 T ) was isolated from wetland soil and characterized. Results of 16S rRNA and draft genome sequence analysis placed strain NA20 T within the genus Terrimonas of the family Chitinophagaceae . Strain NA20 T showed ≤97.1 % sequence similarity to members of the genus Terrimonas and the highest sequence similarity was found to Terrimonas lutea DY T (97.1%). The draft genome of strain NA20 T had a total length of 7 144 125 base pairs. A total of 5659 genes were identified, of which 5613 were CDS and 46 RNA genes were assigned a putative function. Mining the genomes revealed the presence of 225 carbohydrate genes out of 1334 genes. Strain NA20 T contained iso-C 15 : 0 , iso-C 15 : 0 G, iso-C 17 : 0 3-OH and summed feature 3 (C 16 : 1  ω 7 c and/or C 16 : 1  ω 6 c ) as major fatty acids. The predominant quinone was MK-7. The major polar lipids were phosphatidylethanolamine, one unknown polar lipid and one unknown aminophospholipid. Additionally, the functional analysis of NA20 T showed the conversion of protopanaxatriol-mix type major ginsenosides (Rb1, Rc and Rd) to minor ginsenosides F2 and weak conversion of Rh2 and C-K within 24 h. As a result, the genotypic, phenotypic and taxonomic analyses support the affiliation of NA20 T within the genus Terrimonas , for which the name Terrimonas ginsenosidimutans sp. nov. is proposed. The type strain is NA20 T (=KACC 22218 T =LMG 32198 T ).

Four novel bacterial strains, designated as RG327 T , SE158 T , RB56-2 T and SE220 T , were isolated from wet soil in the Republic of Korea. To determine their taxonomic positions, the strains were fully characterized. On the basis of genomic information (16S rRNA gene and draft genome sequences), all four isolates represent members of the genus Sphingomonas . The draft genomes of RG327 T , SE158 T , RB56-2 T and SE220 T consisted of circular chromosomes of 2 226 119, 2 507 338, 2 593 639 and 2 548 888 base pairs with DNA G+C contents of 64.6, 63.6, 63.0 and 63.1 %, respectively. All the isolates contained ubiquinone Q-10 as the predominant quinone compound and a fatty acid profile with C 16 : 0 , C 17 : 1 ω6 c , C 18 : 1 2-OH , summed feature 3 (C 16 : 1 ω7 c /C 16 : 1 ω6c) and summed feature 8 (C 18 : 1 ω 7c /C 18 : 1 ω6 c ) as the major fatty acids, supporting the affiliation of strains RG327 T , SE158 T , RB56-2 T and SE220 T to the genus Sphingomonas . The major identified polar lipids in all four novel isolates were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid and phosphatidylcholine. Moreover, the physiological, biochemical results and low level of DNA-DNA relatedness and average nucleotide identity values allowed the phenotypic and genotypic differentiation of RG327 T , SE158 T , RB56-2 T and SE220 T from other species of the genus Sphingomonas with validly published names and indicated that they represented novel species of the genus Sphingomonas , for which the names Sphingomonas anseongensis sp. nov. (RG327 T = KACC 22409 T = LMG 32497 T ), Sphingomonas alba sp. nov. (SE158 T = KACC 224408 T = LMG 324498 T ), Sphingomonas brevis (RB56-2 T = KACC 22410 T = LMG 32496 T ) and Sphingomonas hankyongi sp. nov., (SE220 T = KACC 22406 T = LMG 32499 T ) are proposed.

A Gram-stain-negative, non-motile, rod-shaped, aerobic and white-coloured bacterium (designated XY19 T ) was isolated from a soil sample of wetland from Godeok Ecological Park, Gangdong-gu, Seoul, Republic of Korea. On the basis of 16S rRNA gene sequencing, strain XY19 T clustered with species of the genus Ramlibacter and appeared closely related to R . ginsenosidimutans DSM 23480 T (98.42 %), R. alkalitolerans JCM 32081 T (97.68 %) and R . monticola JCM 31918 T (97.66 %). The average nucleotide identity between strain XY19 T and three strains ( R. ginsenosidimutans DSM 23480 T , R. alkalitolerans JCM 32081 T and R. monticola JCM 31918 T ) were 80.7, 81.1 and 81.4 %. And the digital DNA-DNA hybridization (dDDH) calculated between strain XY19 T and each of the three strains ( R. ginsenosidimutans DSM 23480 T , R. alkalitolerans JCM 32081 T and R. monticola JCM 31918 T ) were 24.1, 24.4 and 24.5 %. ANI value and dDDH results were a novel species of the genus Ramlibacter . Growth occurs at 10-37 °C on R2A medium in the pressence of 0-1 % NaCl (w/v) and at pH 6.0-8.5. The DNA G+C content of the genomic DNA was 68.7 mol%, and ubiquinone-8 (Q-8) was the major respiratory quinone. The major cellular fatty acids (>5 %) were C 16:1  ω 7 c and/or C 16:1  ω6c (summed feature 3), C 16 : 0 , C 17 : 0 cyclo and C 18:1  ω 7 c and/or C 18:1  ω 6 c (summed feature 8). The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, three unidentified lipids and unidentified aminophospholipid. Physiological and biochemical characteristics indicated that strain XY19 T represents a novel species of the genus Ramlibacter , for which the name Ramlibacter paludis sp. nov. is proposed. The type strain is XY19 T (= KACC 22220 T = LMG 32190 T ).

Background and Objective: The ginsenoside compound K (C-K) (which is a de-glycosylated derivative of major ginsenosides) is effective in the treatment of cancer, diabetes, inflammation, allergy, angiogenesis, aging, and has neuroprotective, and hepatoprotective than other minor ginsenosides. Thus, a lot of studies have been focused on the conversion of major ginsenosides to minor ginsenosides using glycoside hydrolases but there is no study yet published for the bioconversion of minor ginsenosides into another high pharmacological active compound. Therefore, the objective of this study to identify a new gene (besides the glycoside hydrolases) for the conversion of minor ginsenosides C-K into another highly pharmacological active compound., Methods and Results: Lactobacillus brevis which was isolated from Kimchi has showed the ginsenoside C-K altering capabilities. From this strain, a novel potent decarboxylation gene, named HSDLb1, was isolated and expressed in Escherichia coli BL21 (DE3) using the pMAL-c5X vector system. Recombinant HSDLb1 was also characterized. The HSDLb1 consists of 774 bp (258 amino acids residues) with a predicted molecular mass of 28.64 kDa. The optimum enzyme activity was recorded at pH 6.0-8.0 and temperature 30 °C. Recombinant HSDLb1 effectively transformed the ginsenoside C-K to 12-β-hydroxydammar-3-one-20(S)-O-β-D-glucopyranoside (3-oxo-C-K). The experimental data proved that recombinant HSDLb1 strongly ketonized the hydroxyl (-O-H) group at C-3 of C-K via the following pathway: C-K → 3-oxo-C-K. In vitro study, 3-oxo-C-K showed higher solubility than C-K, and no cytotoxicity to fibroblast cells. In addition, 3-oxo-C-K induced the inhibitory activity of ultraviolet A (UVA) against matrix metalloproteinase-1 (MMP-1) and promoted procollagen type I synthesis. Based on these expectations, we hypothesized that 3-oxo-C-K can be used in cosmetic products to block UV radiations and anti-ageing agent. Furthermore, we expect that 3-oxo-C-K will show higher efficacy than C-K for the treatment of cancer, ageing and other related diseases, for which more studies are needed.

A novel Gram-stain-negative, aerobic, rod-shaped, non-motile, cream-coloured strain (G124 T ) was isolated from ginseng soil collected in Yeongju, Republic of Korea. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain G124 T belongs to a distinct lineage within the genus Sphingomonas (family Sphingomonadaceae , order Sphingomonadales and class Alphaproteobacteria ). Strain G124 T was closely related to Sphingomonas rhizophila THG-T61 T (98.5 % 16S rRNA gene sequence similarity), Sphingomonas mesophila SYSUP0001 T (98.3 %), Sphingomonas edaphi DAC4 T (97.6 %) and Sphingomonas jaspsi TDMA-16 T (97.6 %). The strain contained ubiquinone 10 as the major respiratory quinone. The major polar lipid profile of strain G124 T comprised phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine and sphingoglycolipids. The predominant cellular fatty acids of strain G124 T were summed feature 8 (C 18 : 1  ω 7 c /C 18 : 1  ω 6 c ; 33.4 %), summed feature 3 (C 16 : 1  ω 6 c /C 16 : 1  ω 7 c ; 27.2 %) and C 16 : 0 (18.3 %). The genome size of strain G124 T was 2 549 305 bp. The genomic DNA G+C content is 62.0 mol%. The average nucleotide identity and digital DNA-DNA hybridization values between strain G124 T and other Sphingomonas species were in the range of 71.2-75.9 % and 18.7-19.9 %, respectively. Based on the polyphasic analysis such as biochemical, phylogenetic and chemotaxonomic characteristics, strain G124 T represents a novel species of the genus Sphingomonas , for which the name Sphingomonas cremea sp. nov. is proposed. The type strain is G124 T (=KACC 21691 T =LMG 31729 T ).

A Gram-staining-positive, catalase positive and oxidase negative, non-motile, non-flagellated, and oval-shaped bacterium, was designated as I2-34  T , isolated from wetland in Soul South Korea. Colonies were round, entire, raised, and cream colored after two days of incubation on R2A agar plates at 25 °C. Based on genomes (both 16S rRNA gene and draft genome) sequence analysis, strain I2-34  T belongs to the genus Arthrobacter and was most closely related to Arthrobacter deserti YIM CS25 T (98.0%). The strain I2-34  T had a circular genome with length of 5,186,447 base pairs (67 contigs) and 4830 total genes. Out of 4696 were protein-coding genes, 54 tRNA and 4 rRNA genes. The chemotaxonomic analysis indicates iso-C 16:0 , anteiso-C 15:0 , and anteiso - C 17:0 as major fatty acids, phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), and two unidentified glycolipids (GL1, GL2) as major polar lipids. The predominant quinone was MK-8(H 2 ). The peptidoglycan type was A3α with an. L-Lys-L-Ala interpeptide bridge. Thus, the experimental data demonstrated here show that the novel isolate shares the similar major fatty acids, major polar lipid PG, DPG, and GLs, major and major quinone MK8-(H 2 ) with the described members of the genus Arthrobacter. However, the low 16S rRNA gene sequence (98.0%), and some physiological and biochemical characteristics differentiate the I2-34  T from its closest phylogenetic neighbors. As a result, the isolate represents a novel species in within the genus Arthrobacter and family Micrococcaceae for which the name Arthrobacter hankyongi sp. nov. is proposed. The type strain is I2-34  T (= KACC 22217  T , LMG 32197  T )., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Rg3-ginsenoside, a protopanaxadiol saponin, is a well-known adaptogen used for the prevention of cancer and inflammation. However, despite its distinct biological activity, the concentration of Rg3 in the total ginseng extract is insufficient for therapeutic applications. This study aims to convert PPD-class of major ginsenosides into a mixture of minor ginsenoside, to analyze its immune-regulatory role in macrophage cells., Results: Using heat and organic acid treatment, three major ginsenosides, Rc, Rd, and Rb1, were converted into a mixture of minor ginsenosides, GRg3-mix [Rg3(S), Rg3(R), Rg5, and Rk1]. Purity and content analysis of the transformed compound were performed using thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC), compared with their standards. Preceding with the anti-inflammatory activity of GRg3-mix, lipopolysaccharide (LPS)-stimulated murine RAW264.7 macrophage cells were treated with various concentrations of GRg3-mix (6.25, 12.5, 25, and 50 μg/mL). The cell viability assay revealed that the level of cell proliferation was increased, while the nitric oxide (NO) assay showed that NO production decreased dose-dependently in activated RAW264.7 cells. The obtained results were compared to those of pure Rg3(S) ≥ 98% (6.25, 12.5, and 25 μg/mL). Preliminary analysis of the CCK-8 and NO assay demonstrated that GRg3-mix can be used as an anti-inflammatory mediator, but mRNA and protein expression levels were evaluated for further confirmation. The doses of GRg3-mix significantly suppressed the initially upregulated mRNA and protein expression of inflammation-related enzymes and cytokines, namely inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear transcription factor kappa B (NF-κB), tumor necrosis factor (TNF-α), and interleukins (IL-6 and IL1B), as measured by reverse transcription-polymerase chain reaction and western blotting., Conclusions: Our pilot data confirmed that the mixture of minor ginsenosides, namely GRg3-mix, has high anti-inflammatory activity and has an easy production procedure., (© 2023. The Author(s).)

A novel Gram-stain-positive, thin rod-shaped, non-motile, aerobic and creamy-white pigmented bacterium (designated strain WJ7-1 T ) was isolated from activated sludge sampled in Suwon, Republic of Korea. 16S rRNA gene sequence analysis indicated that the isolate belongs to the genus Humibacter , family Microbacteriaceae , with a sequence similarity of 97.9 % to its nearest neighbour Humibacter albus KACC 20986 T . Strain WJ7-1 T contained menaquinone (MK)-10 (16.0 %), 11 (48.5 %) and 12 (35.5 %) as major respiratory quinones. The predominant cellular fatty acids (>15 %) were anteiso-C 17 : 0 , iso-C 16 : 0 and anteiso-C 15 : 0 . The peptidoglycan of strain WJ7-1 T contained the diagnostic diamino acid ornithine and 2,4-diaminobutyric acid alanine. Alanine, glutamic acid and glycine were also present in the cell wall. The characteristic whole-cell sugars present were glucose, galactose, xylose and rhamnose. The polar lipids consisted of phosphatidylglycerol, diphosphatidylglycerol, an unidentified glycolipid and two unidentified phospholipids. Strain WJ7-1 T possessed ginsenoside-converting ( β -glucosidase) activity, which enabled it to transform ginsenoside protopanaxadiol-type Rb 1 (one of the dominant active components of ginseng) to compound K. The genome size of strain WJ7-1 T has 4.2 Mbp and the G+C content is 68.3 mol%. Average nucleotide identity, amino acid identity and digital DNA-DNA hybridization values between strain WJ7-1 T and the closely related strain were 79.8, 36.1 and 23.5 %, respectively, indicating that strain WJ7-1 T represents a novel species of the genus Humibacter . Strain WJ7-1 T could be distinguished from the other members of the genus Humibacter by a number of chemotaxonomic and phenotypic characteristics. Based on polyphasic taxonomic analysis, strain WJ7-1 T (=KACC 19729 T =LMG 30802 T ) represents a novel species within the genus Humibacter , for which the name Humibacter ginsenosidimutans sp. nov. is proposed.

A bacterial strain designated as G188 T was isolated from ginseng field soil in the Republic of Korea. Phylogenetic analysis of 16S rRNA gene sequences showed that strain G188 T formed a distinct lineage within the genus Nocardioides, family Nocardioidaceae , order Propionibacteriales . Sequence similarity revealed that strain G188 T was most closely related to Nocardioides iriomotensis IR27-S3 T (97.7 % 16S rRNA similarity). The genome size of strain G188 T was 4 901 775 bp, and the genomic DNA G+C content was 72.3 mol%. The average nucleotide identity and DNA-DNA hybridization values with other Nocardioides species were less than 75.6 and 20.1 %, respectively. The main fatty acids of strain G188 T were C 17 : 0 , C 17 : 1  ω 8 c and iso-C 16 : 0 . The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylinositol, and the major respiratory quinone was menaquinone 8, supporting that strain G188 T was affiliated with the genus Nocardioides . Based on biochemical, chemotaxonomic and phylogenetic analyses, the novel species Nocardioides panacis G188 T (KACC 21695 T =LMG 31733 T ) is proposed.

The saponins of Bupleurum falcatum L., saikosaponins, are the major components responsible for its pharmacological and biological activities. However, the anti-cancer effects of prosaikogenin and saikogenin, which are glycoside hydrolyzed saikosaponins, are still unknown due to its rarity in plants. In this study, we applied two recombinant glycoside hydrolases that exhibit glycoside cleavage activity with saikosaponins. The two enzymes, BglPm and BglLk, were cloned from Paenibacillus mucilaginosus and Lactobacillus koreensis , and exhibited good activity between 30-37 °C and pH 6.5-7.0. Saikosaponin A and D were purified and obtained from the crude B. falcatum L. extract using preparative high performance liquid chromatography technique. Saikosaponin A and D were converted into saikogenin F via prosaikogenin F, and saikogenin G via prosaikogenin G using enzyme transformation with high β-glycosidase activity. The two saikogenin and two prosaikogenin compounds were purified using a silica column to obtain 78.1, 62.4, 8.3, and 7.5 mg of prosaikogenin F, prosaikogenin G, saikogenin F, and saikogenin G, respectively, each with 98% purity. The anti-cancer effect of the six highly purified saikosaponins was investigated in the human colon cancer cell line HCT 116. The results suggested that saikosaponins and prosaikogenins markedly inhibit the growth of the cancer cell line. Thus, this enzymatic technology could significantly improve the production of saponin metabolites of B. falcatum L.

A Gram-reaction-negative, facultatively aerobic, motile, non-spore-forming, rod-shaped, and denitrifying bacterium, designated dN18-1 T , was isolated from activated sludge, Republic of Korea. This bacterium was investigated via a polyphasic approach to reveal its taxonomic position. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain dN18-1 T belongs to the genus Paludibacterium and is most closely related to P. purpuratum KCTC 42852 T (96.2% sequence similarity), P. yongneupense KACC 11601 T (96.1%), and P. paludis BCRC 80514 T (95.2%). The average nucleotide identity values and digital DNA-DNA hybridization values calculated between strain dN18-1 T and the closely related strains were 72.5-73.1% and 19.0-19.6%. The genome comprises of 3,347,996 bp with a G + C content of 57.3 mol%. Strain dN18-1 T possesses ubiquinone Q-8 as a predominant respiratory quinone, and summed feature 3 (C 16:1 ω6c and/or C 16:1 ω7c), summed feature 8 (C 18:1 ω6c/C 18:1 ω7c), C 16:0 and C 12:0 , as its major fatty acids (>5%). The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified phospholipids and four unidentified aminophospholipids. The results of ANI calculation, digital DNA-DNA hybridization, physiological and biochemical tests allowed phenotypic differentiation of strain dN18-1 T from rephrase other genus Paludibacterium species with validly published names. Therefore, this isolate represents a novel species, for which the name Paludibacterium denitrificans sp. nov. (type strain dN18-1 T  = KACC 19537 T  = CGMCC 1.16961 T ) is proposed., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

A novel Gram-stain-negative, aerobic, yellowish-pigmented, non-motile, rod-shaped bacterial strain, designated strain BO-59 T , was isolated from the activated sludge of a wastewater treatment plant in Hanam City, South Korea. Phylogenetic study based on the 16S rRNA gene sequence positioned BO-59 T in a distinct lineage in the family Chitinophagaceae, sharing less than 92.8% sequence similarity with members of the closely related genera Ferruginibacter, Flavitalea, Pseudoflavitalea, Flavisolibacter, Niastella, and Terrimonas. Phylogenomic- and genomic relatedness analyses revealed that strain BO-59 T is clearly distinguished from other genera in the family Chitinophagaceae by average nucleotide identity < 66.9%) and the genome-to-genome distance (< 29.5%) values. The strain BO-59 T contained MK-7 as the predominant quinone, and iso-C 15:0 , iso-C 17:0 3OH, and iso-C 15:1 G as major fatty acids (> 10%). The DNA G + C content was 39.1 mol% based on genome sequence analysis. The polar lipids of strain BO-59 T were phosphatidylethanolamine, an unidentified aminophospholipid and three unidentified polar lipids. 16S rRNA gene sequence similarity, physiological, and biochemical characteristics indicated that strain BO-59 T represents a novel species of a new genus, for which the name Hanamia caeni gen. nov., sp. nov. is proposed. The type strain is BO-59 T (= KACC 19646 T  = LMG 30865  T )., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

A Gram-stain-negative, non-motile, non-spore-forming, aerobic, rod-shaped and yellow-pigmented bacterium, designated strain Gsoil 183 T , was isolated from ginseng-cultivation soil sampled in Pocheon Province, Republic of Korea. This bacterium was characterized to determine its taxonomic position by using a polyphasic approach. Strain Gsoil 183 T grew at 10-37 °C and at pH 5.0-9.0 on tryptic soy agar. Strain Gsoil 183 T had β -glucosidase activity, which was responsible for its ability to convert ginsenoside Rb1 (one of the dominant active components of ginseng) to F2. Based on 16S rRNA gene sequencing, strain Gsoil 183 T clustered with species of the genus Chryseobacterium and appeared to be closely related to Chryseobacterium sediminis LMG 28695 T (99.1 % sequence similarity), Chryseobacterium lactis NCTC 11390 T (98.6%), Chryseobacterium rhizoplanae LMG 28481 T (98.6%), Chryseobacterium oncorhynchi CCUG 60105 T (98.5%), Chryseobacterium viscerum CCUG 60103 T (98.4%) and Chryseobacterium joostei DSM 16927 T (98.3%). Menaquinone MK-6 was the predominant respiratory quinone and the major fatty acids were iso-C 15 : 0 , iso-C 17 : 0 -3OH and summed feature 3 (C 16 : 1  ω 6 c and/or C 16 : 1  ω 7 c ). The polar lipids were phosphatidylethanolamine, six unidentified glycolipids, five unidentified aminolipids and three unidentified lipids. The G+C content of the genomic DNA was 36.6 mol%. Digital DNA-DNA hybridization between strain Gsoil 183 T and the type strains of C. sediminis , C. lactis , C. rhizoplanae , C. oncorhynchi , C. viscerum and C. joostei resulted in values below 70 %. Strain Gsoil 183 T could be differentiated genotypically and phenotypically from the recognized species of the genus Chryseobacterium . The isolate therefore represents a novel species, for which the name Chryseobacterium panacisoli sp. nov. is proposed, with the type strain Gsoil 183 T (=KACC 15033 T =LMG 23397 T ).

Background: Ginsenoside Rh2( S ) is a promising compound for the prevention of various kinds of cancers, inflammation, and diabetes. However, due to its low concentration (<0.02%), researchers are still trying to find an efficient glycoside hydrolase for the scaled-up production of Rh2( S )., Method: Three glycoside hydrolases (BglBX10, Abf22-3, and BglSk) were cloned in Escherichia coli BL21 (DE3) and the expressed recombinant enzyme was used for the scaled-up production of Rh2( S ) through the conversion of PPD-type (protopanaxadiol) major ginsenosides (Rb1, Rc, and Rd, except Rb2) extracted from Korean red ginseng. Specific and specialized bioconversion pathways were designed that evolved the initial bioconversion of PPD-mix → Rg3( S ) → Rh2( S ). The reaction was started with 50 mg/mL of PPD-mix, 20 mg/mL of BglBX10, Abf22-3, and BglSk in series, respectively. The process was completed in a 10 L jar fermenter with a 5 L working volume at 37 °C for 48 hrs., Results: The designed bioconversion pathways show that Abf22-3 and BglBX10 were responsible for the conversion of Rb1, Rc and Rd → Rg3( S ), and then Rg3( S ) was completely transformed to Rh2( S ) by BglSk. As a result, 15.1 g of ginsenoside Rh2( S ) with 98.0 ± 0.2% purity was obtained after strict purification using the Prep-HPLC system with a 100 φ diameter column. Additionally, BglSk was also investigated for its production activity with seven different kinds of PPD-mix type ginsenosides., Conclusion: Our pilot data demonstrate that BglSk is a suitable enzyme for the gram unit production of ginsenoside Rh2( S ) at the industrial level., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 Published by Elsevier B.V. on behalf of King Saud University.)

The conversion of major ginsenosides into minor ginsenosides attracts a lot of interest because of their biological and pharmaceutical activities. Therefore, for the conversion of ginsenosides, finding a novel competent glycoside hydrolase-producing bacterial strain is useful for future research studies and the mass production of minor ginsenosides. Wastewater samples were collected and screened for novel glycoside hydrolase bacterial strains using Reasoner's 2A+aesculin agar medium. As a result, a novel glycoside hydrolase positive bacterial strain (SB-02 T ) was identified and subjected to a polyphasic taxonomic analysis. Based on genome analysis, strain SB-02 T was found to be affiliated with the family Chitinophagaceae and have less than 92.8 % sequence similarity to other members of the same family. Functional analysis indicated that SB-02 T was able to hydrolyse the ginsenosides Rb1, Rc and Rd to F2 and C-K. Due to the conversion of ginsenosides, the strain's genome was sequenced and the genes were annotated by the NCBI. The average amino acid identity and average nucleotide identity values between SB-02 T and the available reference genomes were 65.7 and 65.9 %, respectively. The novel isolate contained MK-7 as the predominant menaquinone, the major polyamine putrescine, and iso-C 15 : 0 , iso-C 15 : 1 G and iso-C 17 : 0 3-OH as major fatty acids. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Thus, based on the data presented here, strain SB-02 T represents a novel species within a new genus in the family Chitinophagaceae , for which the name Phnomibacter ginsenosidimutans gen. nov., sp. nov. is proposed. The type strain of Phnomibacter ginsenosidimutans is SB-02 T (=KACC 21266 T =LMG 31707 T ). The genome annotation of SB-02 T shows many glycoside hydrolase genes, which may be responsible for the efficient production of many kinds of minor ginsenosides and will be very helpful for future research (target gene cloning) and mass production of either F2 or C-K.

A novel yellow-pigmented bacterial strain (designated BR5-29 T ), was isolated and its taxonomy was studied. Phylogenetic study based on the 16S rRNA and draft genome sequence placed the strain BR5-29 T in a distinct lineage within the family Chitinophagaceae, sharing ≤ 93.4% sequence similarity with members of the closely related genera Ferruginibacter, Flavisolibacter, Flavitalea and Niastella. The novel isolate showed the highest sequence similarity to the genus Ferruginibacter. The draft genome of strain BR5-29 T had a total length of 5,505,520 base pairs. A total of 4585 genes were identified, in which 4537 were CDS and 48 RNA genes were assigned a putative function. The genome annotation of BR5-29 T showed 225 carbohydrate genes which may be responsible for the conversion of major ginsenosides to minor ginsenosides. Strain BR5-29 T contained MK-7 as a predominant quinone, and iso-C 15:0 , iso-C 15:0 G, iso-C 17:0 3-OH, and C 16:1 ω7c and/or C 16:1 ω6c (summed feature 3) as major fatty acids. The polar lipids found in the strain BR5-29 T were phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), five unidentified polar lipids (L1-L5), two unidentified aminolipid and one unidentified aminophospholipid. Our pilot data demonstrate that the novel isolate shares the similar major polar lipid PE, major quinone MK-7 and major fatty acids with the described members of the family Chitinophagaceae. However, the low 16S rRNA gene sequence (< 93.4%), the little high amount of C 12:0 , iso-C 17:0 2-OH and iso-C 15:1 2-OH fatty acids, low DNA G + C content, and the presence of DPG, PG and two unidentified polar lipids (L1 and L3 differentiate the BR5-29 T from its closest phylogenetic neighbors. Thus, the isolate represents a novel genus and species in the family Chitinophagaceae for which the name Ginsengibacter hankyongi gen. nov., sp. nov. is proposed. The type strain is BR5-29 T (= KACC 19446 T  = LMG 30462 T ). Thus, we predict that this novel strain may prove useful for the future research analysis (target gene cloning) and mass production of Rg3.

Two bacterial strains designated as W3-2-3 T and HKS04 T were isolated from mineral water and a soil sample, respectively, in the Republic of Korea. The 16S rRNA genes of the two strains shared a sequence similarity of 93.5 %. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains W3-2-3 T and HKS04 T formed a distinct lineage within the genus Nocardioides of the family Nocardioidaceae (order Propionibacteriales ). The closely related species of strain W3-2-3 T were Nocardioides albidus (98.9 %), Nocardioides caeni (98.8 %), Nocardioides kongjuensis (98.6 %), Nocardioides aromaticivorans (98.5 %), Nocardioides nitrophenolicus (98.4 %), Nocardioides flava (98.2 %) and Nocardioides ginsengisoli (98.1 %). The closest species of strain HKS04 T was Nocardioides halotolerans (98.7 %). The genome sizes of strains W3-2-3 T and HKS04 T were 4741198 and 5 120341 bp, respectively. The genomic DNA G+C contents of strains W3-2-3 T and HKS04 T were 73.3 and 72.1 mol%, respectively. The main fatty acids of strain W3-2-3 T were C 17:1 ω 6 c and iso-C 16:0 and those of strain HKS04 T were iso-C 16:0 and iso-C 16:0 H. The main polar lipids of both strains were diphosphatidylglycerol and phosphatidylglycerol and the predominant respiratory quinone was MK-8(H 4 ), supporting the affiliation of these strains with the genus Nocardioides . Based on the results of biochemical, chemotaxonomic and phylogenetic analyses, two novel species, Nocardioides convexus W3-2-3 T (KACC 21211 T =LMG 31251 T ) and Nocardioides anomalus HKS04 T (KACC 18879 T =LMG 31249 T ), are proposed.

Background: The separation of isomeric compounds from a mixture is a recurring problem in chemistry and phytochemistry research. The purification of pharmacologically active ginsenoside Rb 3 from ginseng extracts is limited by the co-existence of its isomer Rb 2 . The aim of the present study was to develop an enzymatic elimination-combined purification method to obtain pure Rb 3 from a mixture of isomers., Methods: To isolate Rb 3 from the isomeric mixture, a simple enzymatic selective elimination method was used. A ginsenoside-transforming glycoside hydrolase (Bgp2) was employed to selectively hydrolyze Rb 2 into ginsenoside Rd. Ginsenoside Rb 3 was then efficiently separated from the mixture using a traditional chromatographic method., Results: Chromatographic purification of Rb 3 was achieved using this novel enzymatic elimination-combined method, with 58.6-times higher yield and 13.1% less time than those of the traditional chromatographic method, with a lower minimum column length for purification. The novelty of this study was the use of a recombinant glycosidase for the selective elimination of the isomer. The isolated ginsenoside Rb 3 can be used in further pharmaceutical studies., Conclusions: Herein, we demonstrated a novel enzymatic elimination-combined purification method for the chromatographic purification of ginsenoside Rb 3 . This method can also be applied to purify other isomeric glycoconjugates in mixtures., (© 2019 The Korean Society of Ginseng. Publishing services by Elsevier B.V.)

A Gram-reaction negative, aerobic, non-motile, light yellow colored, and rod-shaped bacterium (designated Gr-4 T ) isolated from granules of a wastewater treatment plant, was characterized by a polyphasic approach to clarify its taxonomic position. Strain Gr-4 T was observed to grew optimally at 30 ºC and at pH 7.0 on R2A medium. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain Gr-4 T belongs to the genus Luteimonas of the family Xanthomonadaceae and was most closely related to Luteimonas padinae CDR SL 15 T (99.1%), Luteimonas terricola DSM 22344 T (98.5%) and Luteimonas arsenica 26-35 T (97.6). The genome comprises 2,917,404 bp with a G+C content of 70.5 mol%. The ANI value between strain Gr-4 T and Luteimonas padinae CDR SL 15 T was 87.3%. The DNA-DNA relatedness value between strain Gr-4 T and Luteimonas padinae CDR SL 15 T , Luteimonas terricola DSM 22344 T was 36.4 ± 1.3% and 14.2 ± 1.7%, respectively. The predominant quinone was Q-8. The major fatty acids were iso-C 15:0 , iso-C 16:0 and summed feature 9 (comprising iso-C 17:1 ω9c and/or C 16:0 10-methyl) supported the affiliation of strain Gr-4 T to the genus Luteimonas. Moreover, the physiological, biochemical results, and low level of ANI and DNA-DNA relatedness value allowed the phenotypic and genotypic differentiation of strains Gr-4 T from other Luteimonas species with validly published names. The novel isolate therefore represents a novel species, for which the name Luteimonas granuli sp. nov. is proposed, with the type strain Gr-4 T (=KACC 16614 T  = JCM 18203 T ).

A Gram stain negative, motile, non-spore-forming, rod-shaped, strictly aerobic, beige-pigmented bacterium, designated strain BO-7 T , was isolated from soil of cattle farm, in Seosan, Republic of Korea. On the basis of 16S rRNA gene sequencing, strain BO-7 T clustered with species of the genus Ochrobactrum and appeared closely related to O. haematophilum CCUG 38531 T (98.9%), O. daejeonense KCTC 22458 T (98.1%), O. rhizosphaerae DSM 19824 T (98.1%), O. pituitosum DSM 22207 T (98.0%), and O. pecoris DSM 23868 T (98.0%). The digital DNA-DNA hybridization and average nucleotide identity between strain BO-7 T and the closely related strains were 21.9-39.1%, 78.5-89.5%, respectively, indicating that BO-7 T is a novel species of the genus Ochrobactrum. The DNA G + C content of the genomic DNA was 57.1 mol%, and ubiquinone Q-10 was the predominant respiratory quinone. The polar lipids consisted of phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylmonomethyl-ethanolamine, di-phosphatidylglycerol, the major polyamines were spermidine, putrescine, and sym-homospermidine. The major cellular fatty acids (> 5%) were C 16:0 , C 19:0 cycle ω7c, and C 18:1 ω7c and/or C 18:1 ω6c (summed feature 8). ANI calculation, digital DNA-DNA hybridization, physiological and biochemical characteristics indicated that strain BO-7 T represents a novel species of the genus Ochrobactrum, for which the name Ochrobactrum soli sp. nov. is proposed. The type strain is BO-7 T (= KACC 19676 T  = LMG 30809 T ).

In this study, a novel ginsenoside transforming bacterium, strain W1-2-3 T , was isolated from mineral water. The 16S rRNA gene sequence analysis showed that strain W1-2-3 T shares 93.7-92.2% sequence similarity with the members of the family Sphingomonadaceae and makes a group with Sphingoaurantiacus capsulatus YLT33 T (93.7%) and S. polygranulatus MC 3718 T (93.4%). The novel isolate efficiently hydrolyses the ginsenoside Rc to Rd. The genome comprises a single circular 2,880,809, bp chromosome with 3211 genes in total, and 1993 protein coding genes. The isolate was observed to grow at 10-37 °C and at pH 6-10 on R2A agar medium; maximum growth was found to occur at 25 °C and pH 7.0. Strain W1-2-3 T was found to contain ubiquinone-10 as the predominant quinone and the fatty acids C 16:1 , C 17:1 ω6c, C 14:0 2-OH, summed feature 3 (C 16:1 ω6c/C 16:1 ω7c) and summed feature 8 (C 18 : 1 ω6c/C 18:1 ω7c). The DNA G+C content was determined to be 65.9 mol%. Strain W1-2-3 T can be distinguished from the other members of the family Sphingomonadaceae by a number of chemotaxonomic and phenotypic characteristics. The major polar lipids of strain W1-2-3 T were identified as phosphatidylethanolamine, an unidentified glycolipid and an unidentified polar lipid. The major poly amine was found to be homospermidine. Based on polyphasic taxonomic analysis, strain W1-2-3 T is concluded to represent a novel species within a new genus, for which the name Hankyongella ginsenosidimutans gen. nov., sp. nov. is proposed. The type strain of Hankyongella ginsenosidimutans is W1-2-3 T (= KACC 18307 T  = LMG 28594 T ).

A gram-negative, motile, strictly aerobic, and rod-shaped bacterium designated 176GS2-150 T was isolated from the sponge Hymeniacidon sinapium. The taxonomic position of the novel isolate was confirmed using the polyphasic approach. Strain 176GS2-150 T grew well at 25 °C on marine agar. Based on its 16S rRNA gene sequence, we showed that strain 176GS2-150 T belongs to the family Psychromonadaceae and class Gammaproteobacteria and is related to Corallincola platygyrae JLT2006 T (96.84% sequence similarity). The G + C content of the genomic DNA was 49.0 mol%. The assembled draft genome of strain 176GS2-150 T was 4.2 Mbp and consisted of 14 contigs. The major respiratory quinone was Q-8, and the major fatty acids were summed feature 3 (comprising C 16 :1 ω6c and/or C 16:1 ω7c), summed feature 8 (comprising C 18 :1 ω7c and/or C 18:1 ω6c), C 17:0 iso, C 16:0 , and C 15:0 iso. The polar lipids were phosphatidylglycerol, phosphatidylethanolamine, 3 unidentified phospholipids, and 1 unidentified polar lipid. On the basis of the genotypic and phenotypic characteristics, strain 176GS2-150 T can be placed as a new species within the genus Corallincola; the name Corallincola spongiicola sp. nov. has been proposed, with type strain 176GS2-150 T (= KACC 19890 T  = LMG 31317 T ).

An aerobic and Gram-stain-negative bacterial strain, designated UKS-15 T , was isolated from lake water in the Republic of Korea. Results of 16S rRNA gene sequence and phylogenetic analyses indicated that the novel isolate belongs to the genus Lysobacter and was most closely related to Lysobacter xinjiangensis RCML-52 T (98.0 %), Lysobacter mobilis 9 NM-14 T (97.4 %) and Lysobacter humi FJY8 T (97.2 %). The DNA G+C content was 69.1 mol%. Strain UKS-15 T possessed ubiquinone-8 (Q-8) as the sole respiratory quinone and the fatty acid profile comprised iso-C 15 : 0 , iso-C 17 : 0 and summed feature 9 (iso-C 17 : 1 ω9 c and/or C 16 : 0 10-methyl) as its major components. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and one unidentified aminophospholipid. Moreover, the physiological and biochemical results and low level of DNA-DNA relatedness (<22.0 %) allowed the phenotypic and genotypic differentiation of strain UKS-15 T from other Lysobacter species. Therefore, on the basis of the data from this polyphasic taxonomic study, strain UKS-15 T should represent a novel species of the genus Lysobacter , for which the name Lysobacter lacus sp. nov. is proposed. The type strain is UKS-15 T (=JCM 30983 T =KACC 18719 T ).

In this study, we used a novel α-L-arabinopyranosidase (AbpBs) obtained from ginsenoside-converting Blastococcus saxobsidens that was cloned and expressed in Escherichia coli BL21 (DE3), and then applied it in the biotransformation of ginsenoside Rb 2 into Rd. The gene, termed AbpBs , consisting of 2,406 nucleotides (801 amino acid residues), and with a predicted translated protein molecular mass of 86.4 kDa, was cloned into a pGEX4T-1 vector. A BLAST search using the AbpBs amino acid sequence revealed significant homology with a family 2 glycoside hydrolase (GH2). The over-expressed recombinant AbpBs in Escherichia coli BL21 (DE3) catalyzed the hydrolysis of the arabinopyranose moiety attached to the C-20 position of ginsenoside Rb2 under optimal conditions (pH 7.0 and 40°;C). Kinetic parameters for α-Larabinopyranosidase showed apparent K m and V max values of 0.078 ± 0.0002 micrometer and 1.4 ± 0.1 μmol/min/mg of protein against p -nitrophenyl-α-L-arabinopyranoside. Using a purified AbpBs (1 μg/ml), 0.1% of ginsenoside Rb 2 was completely converted to ginsenoside Rd within 1 h. The recombinant AbpBs could be useful for high-yield, rapid, and low-cost preparation of ginsenoside Rd from Rb 2 .

A Gram-stain-negative, facultatively anaerobic, motile, light-yellow, and rod-shaped bacterium, designated as NY-02 T , was isolated from the sludge of a wastewater treatment plant in Hanam City, Republic of Korea. The comparison of 16S rRNA gene sequences revealed that the strain formed a distinct lineage within the genus Simplicispira and was most closely related to S. suum SC1-8 T (99.0%), S. limi EMB325 T (98.3%), S. psychrophila LMG 5408 T (98.2%), and S. piscis RSG39 T (97.4%). Both average nucleotide identity (ANI) and DNA-DNA hybridization (DDH) values between strain NY-02 T , and its closes type strains [S. suum SC1-8 T , S. limi EMB325 T , S. psychrophila LMG 5408 T , and S. piscis RSG39 T ] were lower than the cut-off (≥ 95-96% for ANI and ≥ 70% for DDH) to define a bacterial species. The genome comprises of 3,709,074 bp with a G + C content of 64.2 mol%. Ubiquinone 8 (Q-8) was the predominant quinone. The major fatty acids were C 16:0 and summed feature 3 (C 16:1 ω7c and/or C 16:1 ω6c), and the major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylethanolamine. The results of the physiological, biochemical, and taxonomic analyses in addition to low ANI and DNA-DNA relatedness values (82.2% and < 34.0%, respectively) indicate that the strain NY-02 T represents a novel species of the genus Simplicispira. The name proposed for strain NY-02 T (= KACC 19731 T  = LMG 31165 T ) is Simplicispira hankyongi sp. nov.

A Gram-stain-negative, strictly aerobic, motile, ivory-coloured and rod-shaped bacterium (designated Gsoil 520 T ) isolated from ginseng cultivation soil was characterized by using a polyphasic approach to clarify its taxonomic position. Strain Gsoil 520 T was observed to grow optimally at 30 °C and pH 7.0 on Reasoner's 2A agar medium. The results of phylogenetic analysis, based on 16S rRNA gene sequence similarities, indicated that Gsoil 520 T belongs to the genus Devosia of the family Hyphomicrobiaceae and was most closely related to Devosia epidermidihirudinis E84 T (98.0 %), Devosia yakushimensis Yak96B T (97.7 %), Devosia neptuniae J1 T (97.7 %) and Devosia chinhatensis IPL18 T (96.8 %). The complete genome of strain Gsoil 520 T is a presumptive circular chromosome of 4 480 314 base pairs having G+C content of 63.7 mol%. A total of 4 354 genes, 4 303 CDS and 43 rRNA genes were assigned a putative function. The major isoprenoid quinone was Q-10. The main polar lipids were phosphatidylglycerol, diphosphatidylglycerol and two unidentified aminolipids (AL1 and AL3). The predominant fatty acids of strain Gsoil 520 T were C 18 : 1 ω7 c 11-methyl, C 16 : 0 and C 18 : 1 ω7 c /C 18 : 1 ω6 c (summed feature 8) supporting the affiliation of strain Gsoil 520 T to the genus Devosia . The low values of DNA-DNA hybridization distinguished strain Gsoil 520 T from the recognized species of the genus Devosia . Thus, the novel isolate represents a novel species of the genus Devosia , for which the name Devosia ginsengisoli sp. nov. is proposed, with the type strain Gsoil 520 T (=KACC 19440 T =LMG 30329 T ).

Background: Several studies have reported that ginsenoside Rg3( S ) is effective in treating metastatic diseases, obesity, and various cancers, however, its presence in white ginseng cannot be estimated, and only a limited amount is present in red ginseng. Therefore, the use of recombinant glycosidases from a Generally Recognized As Safe (GRAS) host strain is a promising approach to enhance production of Rg3( S ), which may improve nutritional activity, human health, and quality of life., Method: Lactobacillus ginsenosidimutans EMML 3041 T , which was isolated from Korean fermented pickle (kimchi), presents ginsenoside-converting abilities. The strain was used to enrich the production of Rg3( S ) by fermenting protopanaxadiol (PPD)-mix-type major ginsenosides (Rb1, Rb2, Rc, and Rd) in four different types of food-grade media (1, MRS; 2, Basel Food-Grade medium; 3, Basel Food-Grade medium-I, and 4, Basel Food-Grade medium-II). Due to its tendency to produce Rg3( S ), the presence of glycoside hydrolase in Lactobacillus ginsenosidimutans was proposed, the whole genome was sequenced, and the probable glycoside hydrolase gene for ginsenoside conversion was cloned., Results: The L. ginsenosidimutans EMML 3041 T strain was whole genome sequenced to identify the target genes. After genome sequencing, 12 sets of glycoside hydrolases were identified, of which seven sets (α,β-glucosidase and α,β-galactosidase) were cloned in Escherichia coli BL21 (DE3) using the pGEX4T-1 vector system. Among the sets of clones, only one clone (BglL.gin-952) showed ginsenoside-transforming abilities. The recombinant BglL.gin-952 comprised 952 amino acid residues and belonged to glycoside hydrolase family 3. The enzyme exhibited optimal activity at 55 °C and a pH of 7.5 and showed a promising conversion ability of major ginsenoside Rb1→Rd→Rg3( S ). The recombinant enzyme (GST-BglL.gin-952) was used to mass produce Rg3( S ) from major ginsenoside Rb1. Scale-up of production using 50 g of Rb1 resulted in 30 g of Rg3( S ) with 74.3% chromatography purity., Conclusion: Our preliminary data demonstrated that this enzyme would be beneficial in the preparation of pharmacologically active minor ginsenoside Rg3( S ) in the functional food and pharmaceutical industries.

A Gram-stain-positive, strictly aerobic, non-motile, non-spore-forming and oval-shaped bacterium, designated strain BO-16 T was isolated from activated sludge. In this study, we describe the taxonomic characterization and classification of this bacterium by using the polyphasic approach. Growth of BO-16 T was observed at 10-40 °C (optimum, 25-37 °C) and at pH 5.0-10.0 (optimum, pH 7.0) on R2A agar. The major fatty acids it contained were iso -C 16:0 , anteiso -C 17:0 and iso -C 17 : 0 and the major polar lipids were diphosphatidylglycerol, phosphatidylinositol and phosphatidylethanolamine. This isoprenoid quinones included MK-8 (H 4 ) and MK-8 (H 6 ). The peptidoglycan contained lysine, serine, alanine, glycine and glutamic acid and represented the peptidoglycan type A4 α . On the basis of 16S rRNA gene sequence similarity, BO-16 T was shown to represent a member of the genus Flexivirga and to be related to Flexivirga oryzae KACC 18597 T (98.4 % sequence similarity), Flexivirga endophytica KCTC 39536 T (97.5 %), Flexivirga alba DSM 24460 T (97.4 %) and Flexivirga lutea KCTC 39625 T (97.3 %). The G+C content of the genomic DNA was 68.0 mol%. The DNA-DNA relatedness values between BO-16 T and its closest phylogenetic neighbours were much lower than 70 %. BO-16 T could be differentiated phylogenetically and phenotypically from the species of the genus Flexivirga with validly published names. Therefore the isolate represents a novel species, for which the name Flexivirga caeni sp. nov. is proposed, with the type strain BO-16 T (=KACC 19647 T =LMG 30859 T ).

A novel Gram-stain-negative, yellowish-pigmented bacterial strain, designated LA-38 T , was isolated from activated sludge of wastewater treatment plants in Hanam city, South Korea. Cell of LA-38 T were rod-shaped, aerobic, motile and non-spore-forming. In phylogenetic analyses based on 16S rRNA genes, LA-38 T clustered with species of the genus Hydrogenophaga and appeared closely related to Hydrogenophaga intermedia DSM 5680 T (99.2 % similarity), Hydrogenophaga palleronii DSM 63 T (98.2 %), Hydrogrenophaga laconesensis KCTC 42478 T (98.1 %), Hydrogenophaga. atypica DSM 15342 T (98.1 %), Hydrogenophaga defluvii DSM 15341 T (98.0 %) and Hydrogenophaga taeniospiralis DSM 2082 T (97.2 %). The average nucleotide identities between LA-38 T and the closely related strains were 79.3-88.5 %, indicating that LA-38 T represents a novel species of the genus Hydrogenophaga . The DNA G+C content of the genomic DNA was 69.9 mol% and ubiquinone Q-8 was the predominant respiratory quinone. The major cellular fatty acids (>5 %) were C 16 : 0 , cyclo-C 19 : 0 , C 16 : 1 ω7 c and/or C 16 : 1 ω6 c (summed feature 3), and C 18 : 1 ω7 c and/or C 18 : 1 ω6 c (summed feature 8). The major polar lipids consisted of phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine, the major polyamines were 2-hydroxyputrescine and putrescine. ANI calculation, physiological and biochemical characteristics indicated that LA-38 T represents a novel species of the genus Hydrogenophaga , for which the name Hydrogenophaga borbori sp. nov. is proposed. The type strain is LA-38 T (=KACC 19730 T =LMG 30805 T ).

A novel Gram-reaction-negative, facultative-aerobic, motile, non-spore-forming, oval-shaped and denitrifying bacterium, designated BO-81 T , was isolated from sludge sampled in the Republic of Korea. This bacterium was investigated via a polyphasic approach to reveal its taxonomic position. The results of phylogenetic analysis based on 16S rRNA gene sequencing indicated that strain BO-81 T belonged to the family Rhodobacteraceae and was related to the closest species Sinirhodobacter ferrireducens (98.8 % sequence similarity), ' Sinorhodobacter hungdaonensis ' (98.4 %), Rhodobacter lacus (97.8 %), Sinorhodobacter populi (96.8 %) and Rhodobacter maris (96.2 %). The average nucleotide identity and DNA-DNA hybridization values between strain BO-81 T and its closely related strains were 77.7-94.2 % and 20.1-55.9 %, respectively, indicating that BO-81 T represents a novel species of the genus Sinirhodobacter . Growth occurred at 18-40 °C on Reasoner's 2A medium in the presence of 0-7 % NaCl (w/v) and at pH 6.0-9.0. Strain BO-81 T was characterized chemotaxonomically as having ubiquinone 10 as its predominant respiratory quinone, summed feature 8 (C 18 : 1 ω 6 c and/or C 18 : 1 ω 7 c ) as its major fatty acid and phosphatidylglycerol and phosphatidylethanolamine as its predominant polar lipids. The G+C content of the genomic DNA was 68.3 mol%. The results of physiological and biochemical tests allowed phenotypic differentiation of strain BO-81 T from other Sinirhodobacter species with validly published names. Therefore, the isolate represented a novel species, for which the name Sinirhodobacter hankyongi sp. nov. (type strain BO-81 T =KACC 19677 T =LMG 30808 T ) is proposed.

A Gram-stain-negative, rod-shaped, motile, facultatively aerobic and ivory-pigmented bacterium (designated strain LA-55 T ) was isolated from a river in the Republic of Korea. On the basis of 16S rRNA gene sequencing, strain LA-55 T clustered with species of the genus Brevundimonas and was closely related to B revundimonas kwangchunensis KSL-102 T (97.3 %), B revundimonas aurantiaca DSM 4731 T (97.1 %), B revundimonas albigilva NHI-13 T (97.0 %), B revundimonas balnearis FDRGB2b T (97.0 %) and Brevundimonas aveniformis DSM 17977 T (97.0 %). The average nucleotide identity value between strain LA-55 T and its closest-related strain was 74.1 %, indicating that strain LA-55 T represents a novel species of the genus Brevundimonas . Growth occurred at 15-40 °C on Reasoner's 2A medium in the presence of 0-2 % NaCl (w/v) and at pH 6.0-8.0. The genomic DNA G+C content was 70.5 mol% and ubiquinone 10 (Q-10) was the major respiratory quinone. The major cellular fatty acids (>5 %) were C 1 8 :1 ω 6 c and/or C 1 8 :1 ω 7 c (summed feature 8), C 16 : 0 , C 1 6 :1 ω 6 c and/or C 1 6 :1 ω 7 c (summed feature 3) and C 18 : 1 ω 7 c 11-methyl. The polar lipids consisted of phosphatidylglycerol, 1,2-di- O -acyl-3- O -[d-glucopyranosyl-(1→4)- α -d-glucopyranuronosyl]glycerol, 1,2-di- O -acyl-3- O-α -d-glucopyranuronosyl glycerol, unidentified aminolipid, unidentified phosphoglycolipid and unidentified lipids. Physiological and biochemical characteristics indicated that strain LA-55 T represents a novel species of the genus Brevundimonas , for which the name Brevundimonas fluminis sp. nov. is proposed. The type strain is LA-55 T (=KACC 19639 T =LMG 30850 T ).

Use of recombinant glycosidases is a promising approach for the production of minor ginsenosides, e.g., Compound K (CK) and F 1 , which have potential applications in the food industry. However, application of these recombinant enzymes for food-grade preparation of minor ginsenosides are limited by the lack of suitable expression hosts and low productivity. In this study, Corynebacterium glutamicum ATCC13032, a GRAS strain that has been used extensively for the industrial-grade production of additives for foodstuffs, was employed to express a novel β-glucosidase (MT619) from Microbacterium testaceum ATCC 15829 with high ginsenoside-transforming activity. A cellulose-binding module was additionally fused to the N-terminus of MT619 for immobilization on cellulose, which is an abundant and safe material. Via one-step immobilization, the fusion protein in cell lysates was efficiently immobilized on regenerated amorphous cellulose at a high density (maximum 984 mg/g cellulose), increasing the enzyme concentration by 286-fold. The concentrated and immobilized enzyme showed strong conversion activities against protopanaxadiol- and protopanaxatriol-type ginsenosides for the production of CK and F 1 . Using gram-scale ginseng extracts as substrates, the immobilized enzyme produced 7.59 g/L CK and 9.42 g/L F 1 in 24 h. To the best of our knowledge, these are the highest reported product concentrations of CK and F 1 , and this is the first time that a recombinant enzyme has been immobilized on cellulose for the preparation of minor ginsenosides. This safe, convenient, and efficient production method could also be effectively exploited in the preparation of food-processing recombinant enzymes in the pharmaceutical, functional food, and cosmetics industries.

In this study, many bacterial strains were screened for the production of minor ginsenosides, but based on conversion competence among the strains, the strain Niabella ginsenosidivorans BS26 T has the good ginsenoside-transforming ability. Therefore, the strain BS26 T was selected for complete genome sequence analysis to determine the target (glycoside hydrolase) functional genes. Whole genome analysis of strain BS26 T showed 43 glycoside hydrolase genes in total. To determine the target functional gene, 12 sets of six different glycoside hydrolases (3 set of β-glucosidase; 3 set of trehalase; 3 set of arabinofuranosidase; 2 set of xylosidase; and one set of each α-galactosidase and α-fucosidase, respectively) were selected and cloned in E. coli BL21 (DE3) using the pGEX4T-1 vector and were characterized. Among these 12 sets of clones, only one, β-glucosidase (BglNg-767), showed ginsenoside conversion ability. The BglNg-767 comprised 767 amino acids and belonged to glycoside hydrolase family 3 (GH3). The recombinant GST-BglNg-767 was capable of altering the ginsenosides Rb 1 , Rd, and gypenoside XVII (Gyp-XVII) to F 2 ; Rb 2 to C-O; Rb 3 to C-Mx1, and Rc to C-Mc1. Besides, complete genome sequence analysis of strain BS26 T also indicates 30 endopeptidase genes, which may be responsible for self-hydrolysis of the proteins. Therefore, using SDS-PAGE analysis, we predict that the difference between the molecular weight of the expressed protein (around 90 kDa) and the predicted amino-acid sequence (102.7 kDa) is due to self-hydrolysis of the proteins., Competing Interests: Conflict of interestThe authors declare that there are no conflict of interest.

A Gram-stain-negative, aerobic, rod-shaped bacterium, designated strain C8-1 T , was isolated from the rhizosphere soil of Nicotiana tabacum L. collected from Kunming, south-west China. The cells showed oxidase-positive and catalase-positive reactions. Growth was observed at 10-40 °C, at pH 6.0-8.0 and in the presence of up to 1 % (w/v) NaCl, with optimal growth at 30 °C and pH 7.0. The predominant isoprenoid quinone was Q-8. The major fatty acids were identified as iso-C15 : 0, iso-C17 : 0 and iso-C17 : 1ω9c. The cellular polar lipids contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmethylethanolamine, five unidentified phospholipids and two unidentified aminophospholipids. The genomic DNA G+C content was 70.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain C8-1 T should be assigned to the genus Lysobacter. 16S rRNA gene sequence similarity analysis showed that strain C8-1 T was closely related to Lysobacter cavernae YIM C01544 T (98.6 %), Lysobacter soli DCY21 T (97.6 %), Lysobacter panacisoli CJ29 T (97.3 %), Lysobacter firmicutimachus PB-6250 T (97.3 %), Lysobacter niastensis GH41-7 T (97.3 %) and Lysobacter gummosus KCTC 12132 T (97.1 %). DNA-DNA hybridization data indicated that the isolate may represent a novel genomic species belonging to the genus Lysobacter. Polyphasic taxonomic characteristics indicated that strain C8-1 T represents a novel species of the genus Lysobacter, for which the name Lysobacter tabacisoli sp. nov. is proposed. The type strain is C8-1 T (=KCTC 62034 T =CGMCC 1.16271 T ) .

A newly isolated bacterial strain designated as HKS19 was isolated from a ginseng cultivation soil sample collected in South Korea. Cells of the strain HKS19 were Gram-stain-negative, rod, oval-shaped and they formed yellow colonies when grown on R2A agar at 30 °C. HKS19 showed the highest 16S rRNA gene sequence similarity (98.6%) with Sphingomonas asaccharolytica NBRC 15499 T . Its growth was observed at 10-37 °C (optimum 30 °C), pH 6-9 (optimum pH 7), and in the presence of 0-1% NaCl (optimum 0%). The genome size of HKS19 was 3.4 Mb and the G+C content was 65.1 mol%. The main polar lipid of strain HKS19 was diphosphatidylglycerol (DPG), the predominant respiratory quinone was Q-10 and the major fatty acids were a summed feature 8 (C 18 : 1 ω6 c / C 18 : 1 ω7 c) and C 16 : 0 . Based on the phylogenetic, genotypic, phenotypic and chemotaxonomic analysis, strain HKS19 represents a newly isolated species of the genus Sphingomonas , for which the name Sphingomonas panacisoli is proposed. The type strain is HKS19 T (=KACC 18881 T =LMG 29564 T ).

A Gram-stain-negative, strictly aerobic, non-motile, ivory colored and rod-shaped bacterium (designated Gsoil 652 T ) isolated from ginseng cultivating soil, was characterized using a polyphasic approach to clarify its taxonomic position. Strain Gsoil 652 T was observed to grow optimally at 30 °C and at pH 7.0 on R2A agar medium. Phylogenetic analysis, based on 16S rRNA gene sequences similarities, indicated that Gsoil 652 T belongs to the genus Caballeronia of the family Burkholderiaceae and was most closely related to Caballeronia choica LMG 22940 T (98.9%), Caballeronia udeis LMG 27134 T (98.9%), Caballeronia sordidicola LMG 22029 T (98.2%) and Caballeronia humi LMG 22934 T (98.1%). The DNA G+C content was 62.1 mol% and Q-8 was the major isoprenoid quinone. The main polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, unidentified aminophospholipid, and unidentified phospholipid. The predominant fatty acids were C 16:0 , C 17:0 cyclo and C 19:0 cyclo ω8c. The DNA-DNA relatedness value between strain Gsoil 652 T and closely related type strains of Caballeronia species were less than 36.0%. Moreover, strain Gsoil 652 T could be distinguished phenotypically from the recognized species of the genus Caballeronia. The novel isolate, therefore, represents a novel species, for which the name Caballeronia ginsengisoli sp. nov. is proposed, with the type strain Gsoil 652 T (= KACC 19441 T  = LMG 30326 T ).

Background: Notoginseng stem-leaf (NGL) ginsenosides have not been well used. To improve their utilization, the biotransformation of NGL ginsenosides was studied using ginsenosidase type-I from Aspergillus niger g.848., Methods: NGL ginsenosides were reacted with a crude enzyme in the RAT-5D bioreactor, and the dynamic changes of multi-ginsenosides of NGL were recognized by HPLC. The reaction products were separated using a silica gel column and identified by HPLC and NMR., Results: All the NGL ginsenosides are protopanaxadiol-type ginsenosides; the main ginsenoside contents are 27.1% Rb3, 15.7% C-Mx1, 13.8% Rc, 11.1% Fc, 7.10% Fa, 6.44% C-Mc, 5.08% Rb2, and 4.31% Rb1. In the reaction of NGL ginsenosides with crude enzyme, the main reaction of Rb3 and C-Mx1 occurred through Rb3→C-Mx1→C-Mx; when reacted for 1 h, Rb3 decreased from 27.1% to 9.82 %, C-Mx1 increased from 15.5% to 32.3%, C-Mx was produced to 6.46%, finally into C-Mx and a small amount of C-K. When reacted for 1.5 h, all the Rb1, Rd, and Gyp17 were completely reacted, and the reaction intermediate F2 was produced to 8.25%, finally into C-K. The main reaction of Rc (13.8%) occurred through Rc→C-Mc1→C-Mc→C-K. The enzyme barely hydrolyzed the terminal xyloside on 3- O - or 20- O -sugar-moiety of the substrate; therefore, 9.43 g C-Mx, 6.85 g C-K, 4.50 g R7, and 4.71 g Fc (hardly separating from the substrate) were obtained from 50 g NGL ginsenosides by the crude enzyme reaction., Conclusion: Four monomer ginsenosides were successfully produced and separated from NGL ginsenosides by the enzyme reaction.

A Gram-stain-negative, non-spore-forming, facultative, rod-shaped bacterium (designated LA-28 T ) was isolated from a sludge sample from a wastewater treatment plant in Hanam city, Republic of Korea. On the basis of 16S rRNA gene sequencing, strain LA-28 T clustered with species of the genus Mesorhizobium and appeared closely related to M. jarvisii LMG 28313 T (96.8%), M. waimense ICMP 19557 T (96.7%), and M. huakuii LMG 14107 T (96.7%). Growth occurs at 18-40°C on R2A medium in the presence of 1-4% NaCl (w/v) and at pH 6-8. The DNA G+C content was 61.2 mol%, and the predominant quinone was ubiquinone-10 (Q-10). The major cellular fatty acids (> 5%) were C 16:0 , C 19:0 ω8c cyclo, C 18:1 ω7c 11-methyl, and C 18:1 ω7c and/or C 18:1 ω6c (summed feature 8). Major polar lipids were phosphatidylglycerol (PG), phosphatidylethanolamine (PE), phosphatidyl-N-methylethanolamine (PME), and phosphatidylcholine (PC). Physiological and biochemical characteristics indicated that strain LA-28 T represents a novel species of the genus Mesorhizobium, for which the name Mesorhizobium denitrificans sp. nov. is proposed. The type strain is LA-28 T (= KACC 19675 T = LMG 30806 T ).

Neuroblastoma (NB) is the most common childhood cancer, with a very poor prognosis. More than 60% of children with NB die within five years; therefore, a more effective therapy for NB is required. Although ginsenoside has been shown to significantly inhibit the growth of various cancers, the effect of ginsenoside Rk1 on neuroblastoma has not been known yet. Hence, we examined the anticancer effects of highly pure Rk1 on neuroblastoma cell lines. The apoptotic effects of Rk1 on neuroblastoma cells were examined using cell viability assay, flow cytometry and cell staining assay, and the change in gene expression levels were analysed using RT-PCR, western blots, and immunohistochemistry. The metastatic effect of Rk1 was monitored by wound healing assay, invasion and migration with Matrigels. Rk1 inhibited neuroblastoma cell viability dose-dependently. Rk1-induced apoptosis was investigated through nuclear condensation and mitochondrial membrane potential loss, and it showed that Rk1 can induce cell cycle arrest at the G0/G1 phase but also inhibit the metastatic ability of neuroblastoma cells. Moreover, Rk1 (30 mg/kg) injections markedly inhibited xenograft tumor growth. These findings demonstrate that Rk1 might be valuable in the development of anti-cancer agents for neuroblastoma treatment.

A Gram-negative, aerobic, non-motile, non-spore-forming and rod-shaped bacterial strain, designated SK3863 T , was isolated from rotten biji (residue remaining after making tofu). This bacterium was characterized in order to determine its taxonomic position by using the polyphasic approach. Strain SK3863 T grew well at 25-37 °C on Reasoner's 2A agar plates. On the basis of 16S rRNA gene sequence similarity, strain SK3863 T belonged to the family Comamonadaceae and was related to Ottowia beijingensis GCS-AN-3 T (96.5 % sequence similarity) and Ottowia pentelensis RB3-7 T (96.4 %). Lower sequence similarities (96.2 %) were found to all of the other recognized members of the genus Ottowia. The G+C content of the genomic DNA was 65.8 mol%. The major respiratory lipoquinone was ubiquinone 8 and the major fatty acids were C16 : 1ω6c/C16 : 1ω7c, C16 : 0 and C18 : 1ω7c/C18 : 1ω6c. Strain SK3863 T could be differentiated genotypically and phenotypically from the recognized species of the genus Ottowia. The isolate therefore represents a novel species, for which the name Ottowia konkukae sp. nov. is proposed, with the type strain SK3863 T (=KCCM 43236 T =DSM 105395 T ).

A Gram-negative, non-spore-forming and rod-shaped, bacterium (designated Gsoil 531 T ) was isolated from soil of a ginseng field. On the basis of 16S rRNA gene sequence, strain Gsoil 531 T clustered with species of the genus Mesorhizobium and was closely related to M. camelthorni CCNWXJ 40-4 T (98.9%) and M. alhagi CCNWXJ12-2 T (98.7%). The DNA G + C content was 62.9 mol% and the predominant quinone was ubiquinone-10 (Q-10). The major cellular fatty acids were C 16:0 , C 19:0 cyclo ω8c and summed feature 8 (C 18:1 ω7c/C 18:1 ω6c). The DNA-DNA hybridization values were less than 35.0% between novel isolate and its closest reference strains M. camelthorni HAMBI 3020 T , M. alhagi HAMBI 3019 T and M tamadayense LMG 26736 T . Physiological, biochemical and low values of DNA-DNA hybridization results enabled strain Gsoil 531 T to be differentiated genotypically and phenotypically from all known species of the genus Mesorhizobium. Therefore, strain Gsoil 531 T signifies a novel species of the genus Mesorhizobium, for which the name Mesorhizobium hankyongi sp. nov. is proposed. The type strain Gsoil 531 T (= KACC 19443 T  = LMG 30463 T ).

An obligate anaerobic, Gram-reaction-negative, non-sporeforming, non-motile, rod shaped bacterium designated YMC B3181 T was isolated from the blood of a patient with peritonitis. Strain B3181 T grew at 20 to 40°C with optimum growth at 37°C. 16S rRNA gene sequence similarity showed strain B3181 T belongs to the genus Parabacteroides and is closely related to Parabacteroides faecis 157 T (97.3%), Parabacteroides gordonii MS-1 T (96.6%), and Parabacteroides goldsteinii DSM 19448 T (95.7%). The G + C content of the genomic DNA was 42.3 mol%. The major cellular fatty acids were anteiso-C 15:0 and iso-C 17:0 3-OH, and the predominant respiratory quinones were MK-9 and MK-10 menaquinones. Genomic and chemotaxonomic data supported affiliation of B3181 T with the genus Parabacteroides. Strain B3181 T was phylogenetically and phenotypically different from recognized species of the genus Parabacteroides. Accordingly, this isolate belongs to a novel species for which the name Parabacteroides chongii sp. nov. (type strain YMC B3181 T = LMG 30065 T = KACC 19034 T ) is proposed.

A Gram-stain negative, aerobic, motile, non-spore-forming and rod-shaped bacterial strain, designated YIM 730227 T , was isolated from a soil sample, collected from Karak district, Khyber-Pakhtun-Khwa, Pakistan. The bacterium was characterized using a polyphasic taxonomic approach. Pairwise comparison of the 16S rRNA gene sequences showed that strain YIM 730227 T is closely related to Phenylobacterium lituiforme FaiI3 T (97.5% sequence similarity), Phenylobacterium muchangponense A8 T (97.4%), Phenylobacterium panacis DCY109 T (97.1%), Phenylobacterium immobile E T (97.1%) and Phenylobacterium composti 4T-6 T (97.0%), while also sharing 98.0% sequence similarity with Phenylobacterium hankyongense HKS-05 T after NCBI blast, showing it represents a member of the family Caulobacteraceae. The major respiratory quinone was Q-10 and the major fatty acids were C 16:0, summed feature 8 (comprising C 18:1 ω7c and/or C 18:1 ω6c), C 18:1 ω7c 11-methyl and C 17:0 . The polar lipids were phosphatidylglycerol, unidentified glycolipids, phospholipid and unidentified lipid. The G + C content of the genomic DNA was 68.2 mol%. The DNA-DNA relatedness values of strain YIM 730227 T with P. hankyongense HKS-05 T , P. lituiforme FaiI3 T , P. muchangponense A8 T , P. panacis DCY109 T , P. immobile E T and P. composti 4T-6 T were 31.3 ± 0.6, 26.1 ± 0.2, 24.3 ± 0.1, 21.8 ± 0.9, 19.8 ± 0.6 and 18.2 ± 1.1%, respectively, values lower than 70%. Besides the morphological and chemotaxonomic characteristics, phylogenetic analyses of 16S rRNA gene sequences and the biochemical characteristics indicated that the strain YIM 730227 T represents a novel member of the genus Phenylobacterium, for which the name Phenylobacterium terrae sp. nov. (type strain YIM 730227 T = KCTC62324 T  = CGMCC 1.16326 T ) is proposed.

Gram-positive, aerobic, non-motile, pale-yellow, and rodshaped bacterium, designated as Gsoil 188 T , was isolated from the soil of a ginseng field in Pocheon, South Korea. A phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that the strain formed a distinct lineage within the genus Brevibacterium and was most closely related to B. epidermidis NBRC 14811 T (98.4%), B. sediminis FXJ8.269 T (98.2%), B. avium NCFB 3055 T (98.1%), and B. oceani BBH7 T (98.1%), while it shared less than 98.1% identity with the other species of this genus. The DNA G + C content was 68.1 mol%. The predominant quinone was MK-8(H 2 ). The major fatty acids were anteiso-C 15:0 and anteiso-C 17:0 . The cell wall peptidoglycan of strain Gsoil 188 T contained meso-diaminopimelic acid. The major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, and an unidentified aminolipid. The physiological and biochemical characteristics, low DNA-DNA relatedness values, and taxonomic analysis allowed the differentiation of strain Gsoil 188 T from the other recognized species of the genus Brevibacterium. Therefore, strain Gsoil 188 T represents a novel species of the genus Brevibacterium, for which the name Brevibacterium anseongense sp. nov. is proposed, with the type strain Gsoil 188 T (= KACC 19439 T = LMG 30331 T ).

Ginsenoside Rh 2 , a protopanaxadiol saponin from ginseng, has been reported to have strong anti-inflammatory activity. However, the concentration of ginsenoside Rh 2 is very low (>0.001%) in the total ginseng extracted, which is not enough for production despite its high pharmacological effects. Thus, in this study, we evaluated the anti-inflammatory effect of ginsenoside Rh 2 -mix (GRh 2 -mix) on lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophage cells. From the high-performance liquid chromatography analysis, it was confirmed that the GRh 2 -mix was mainly composed of 20(S)-Rh 2 , 20(R)-Rh 2 , Rk 2 , and Rh 3 . The LPS-stimulated RAW 264.7 cells were treated with different concentrations of GRh 2 -mix (100, 200, 400, 500 μg/mL). The cell counting kit-8 assay showed that the GRh 2 -mix treatment increased cell proliferation in LPS-stimulated RAW 264.7 murine macrophage cells. The GRh 2 -mix inhibited nitric oxide production in a dose-dependent manner, suggesting an anti-inflammatory effect. Furthermore, reverse transcription polymerase chain reaction and Western blot results also indicated that the GRh 2 -mix suppressed inflammatory genes such as iNOS, TNF-α, COX-2, IL-1β, IL-6, and NF-κB. In summary, these results suggest that the GRh 2 -mix exhibits anti-inflammatory activity via the downregulation of the NF-κB pathway and has high efficiency with a simple production procedure.

A Gram-positive, rod-shaped, non-spore-forming, and aerobic bacterium (Gsoil 556 T ) was isolated from soil of a ginseng field and subjected to its taxonomic position. Based on 16S rRNA gene sequence similarity, strain Gsoil 556 T was shown to belong to the genus Actinomadura of the family Thermomonosporaceae and was closely related to A. montaniterrae CYP1-1B T (99.3%), A. nitritigenes DSM 44137 T (98.7%), and A. rudentiformis HMC1 T (98.5%), while it showed less than 98.4% sequence similarity to the other species of this genus. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that it is most closely related to A. rudentiformis HMC1 T and A. nitritigenes DSM 44137 T . The DNA G+C content was 73.1 mol%. The peptidoglycan was meso-diaminopimelic acid and the whole-cell sugar contained fucose, galactose, glucose, mannose, and ribose. The predominant menaquinone (KK) was MK-9(H 8 ) [55%] and MK-9(H 6 ) [45%]. The major cellular fatty acids were C 14:0 , C 16:0 , C 18:1 ω9c and summed feature 3 (C 16:1 ω6c/C 16:1 ω7c). All these data supported the affiliation of strain Gsoil 556 T to the genus Actinomadura. The DNA-DNA hybridization between strain Gsoil 556 T and its phylogenetically closest relatives were less than 40%. Furthermore, the results of physiological and biochemical tests enabled strain Gsoil 556 T to be differentiated genotypically and phenotypically from currently known Actinomadura species. Therefore, strain Gsoil 556 T represents a novel species of the genus Actinomadura, for which the name Actinomadura hankyongense sp. nov. is proposed. The type strain Gsoil 556 T (=KACC 19438 T =LMG 30327 T ).