Chang-Tian Li

Activation Effects of Polysaccharides of Flammulina velutipes Mycorrhizae on the T Lymphocyte Immune Function

Flammulina velutipes mycorrhizae have increasingly been produced with increasing of F. velutipes production. A mouse model was thus used to examine potential effect of F. velutipes mycorrhizae on the immune function. Fifty female Wistar mice (5-weeks-old) weighed 15–20 g were randomly allocated into five groups. Polysaccharide of F. velutipes mycorrhizae were treated with mice and mice spleen lymphocytes. The levels of CD3+, CD4+, and CD8+ T lymphocyte, interleukin-2 (IL-2), and tumor necrosis factor-a (TNF-α) were determined. The results showed that the proportions of CD3+, and CD4+ T lymphocyte, the ratio of CD4+/CD8+, and the levels of IL-2 and TNF-a were significantly increased in polysaccharide of F. velutipes mycorrhizae, while the proportion of CD8+ T lymphocyte was decreased in polysaccharide of F. velutipes mycorrhizae-dose dependent manner. Our findings indicated that a long term exposure of polysaccharide of F. velutipes mycorrhizae could activate the T lymphocyte immune function. Polysaccharide of F. velutipes mycorrhizae was expected to develop into the immune health products.

Paracoccus hibisci sp. nov., isolated from the rhizosphere of Hibiscus syriacus L. (Mugunghwa flower)

A Gram-stain-negative, aerobic, short-rod-shaped bacterium, motile by means of one flagellum (THG-T2.8T), was isolated from the rhizosphere of Mugunghwa flower. Growth occurred at 10-37 °C (optimum 28 °C), at pH 6-8 (optimum 7) and with 0-5 % NaCl (optimum 1 %). The major quinone was ubiquinone-10 (Q-10). The major fatty acids were C10 : 0 3-OH, C16 : 0, C18 : 0 and C18 : 1ω7c. The polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylglycerol, one unidentified aminolipid, two unknown phospholipids, one unknown glycolipid and one unidentified lipid. The DNA G+C content of strain THG-T2.8T was 65.5 mol%. Based on 16S rRNA gene sequence analysis, the nearest phylogenetic neighbours of strain THG-T2.8T were identified as Paracoccus tibetensis Tibet-S9a3T (98.6 %), Paracoccus aestuarii B7T (98.4 %), Paracoccus rhizosphaerae CC-CCM15-8T (98.3 %) and Paracoccus beibuensis JLT1284T (98.2 %). Levels of sequence similarity among strain THG-T2.8T and other species of the genus Paracoccus were lower than 98.0 %. DNA-DNA hybridization values between strain THG-T2.8T and P. tibetensis Tibet-S9A3TT, P. aestuarii B7T, P. rhizosphaerae CC-CCM15-8T and P. beibuensisJLT1284T were 36.5 % (38.8 %, reciprocal analysis), 32.8 % (34.8 %), 31.6 % (33.8 %) and 15.3 % (24.8 %), respectively. On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain THG-T2.8T represents a novel species of the genus Paracoccus, for which the name Paracoccus hibisci sp. nov. is proposed. The type strain is THG-T2.8T (=KACC 18932T=CCTCC AB 2016181T).

Inhibitory and Acceleratory Effects of Inonotus obliquus on Tyrosinase Activity and Melanin Formation in B16 Melanoma Cells

The aim of the present study is to preliminarily investigate the antimelanogenesis effect of Inonotus obliquus extracts by cell-free mushroom tyrosinase assay. It was found that petroleum ether and n-butanol extracts might contain unknown potential tyrosinase inhibitors, while its ethyl acetate extract might contain some unknown accelerators. Six compounds were isolated and their structures were identified by interpretation of NMR data and nicotinic acid was first discovered in Inonotus obliquus. In cells testing, betulin and trametenolic acid decreased tyrosinase activity and melanin content, while inotodiol and lanosterol significantly increased tyrosinase activity and melanin content, showing an AC⁡50 of 9.74 and 8.43 μM, respectively. Nicotinie acid, 3β,22,25-trihydroxy-lanosta-8-ene, had a little or no effect on tyrosinase. Betulin exhibited a mode of noncompetitive inhibition with a K I = K IS of 0.4 μM on tyrosinase activity showing an IC50 of 5.13 μM and being more effective than kojic acid (6.43 μM), and trametenolic acid exhibited a mode of mixed inhibition with a K I of 0.9 μM, K IS of 0.5 μM, and an IC50 of 7.25 μM. We proposed betulin and trametenolic acid as a new candidate of potent tyrosinase inhibitors and inotodiol and lanosterol as accelerators that could be used as therapeutic agent.

Paracoccus hibiscisoli sp. nov., isolated from the rhizosphere of Mugunghwa (Hibiscus syriacus)

A Gram-reaction-negative, aerobic, non-motile, short-rod-shaped bacterium (THG-T2.31T) was isolated from the rhizosphere of Mugunghwa (Hibiscus syriacus). Growth occurred at 10-35 °C (optimum 28 °C), at pH 5.0-8.0 (optimum pH 7.0) and with 0-4.0 % NaCl (optimum 1.0 %). Based on 16S rRNA gene sequence analysis, the nearest phylogenetic neighbours of strain THG-T2.31T were identified as Paracoccus marcusii DSM 11574T (98.4 %), Paracoccus haeundaensis BC74171T (98.3 %), Paracoccus carotinifaciens E-396T (98.3 %), Paracoccus aestuarii B7T (97.3 %) and Paracoccus seriniphilus MBT-A4T (97.0 %); levels of similarity with the type strains of other species of the genus Paracoccus were lower than 97.0 %. The polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, one unidentified aminolipid and two unidentified phospholipids. The major fatty acids were C16 : 0, C18 : 0, C10 : 0 3-OH, and C18 : 1ω7c. The quinone was ubiquinone-10 (Q-10). The DNA G+C content of strain THG-T2.31T was 69.1 mol%. DNA-DNA hybridization values between strain THG-T2.31T and P. marcusii DSM 11574T, P. haeundaensis BC74171T, P. carotinifaciens E-396T, P. aestuarii B7T and P. seriniphilus MBT-A4T were 38.9 % (34.9 %, reciprocal analysis), 29.1 % (23.5 %), 28.0 % (19.7 %), 18.9 % (9.3) and 13.1 % (6.2 %). On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain THG-T2.31T represents a novel species of the genus Paracoccus, for which the name Paracoccus hibiscisoli sp. nov. is proposed. The type strain is THG-T2.31T (=KACC 18933T=CCTCC AB 2016182T).

Actinotalea solisilvae sp. nov., isolated from forest soil and emended description of the genus Actinotalea

A Gram-stain-positive, aerobic, non-motile and short-rod-shaped actinobacterium, designated THG-T121T, was isolated from forest soil. Growth occurred at 10-40 °C (optimum 28-30 °C), at pH 6-8 (optimum 7) and at 0-4 % NaCl (optimum 1 %). Based on 16S rRNA gene sequence analysis, the nearest phylogenetic neighbours of strain THG-T121T were identified as Actinotalea ferrariae KCTC 29134T (97.9 %), Actinotalea fermentans KCTC 3251T (97.3 %), Cellulomonas carbonis KCTC 19824T (97.2 %). 16S rRNA gene sequence similarities among strain THG-T121T and other recognized species were lower than 97.0 %. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, two phosphatidylinositol mannosides, one unidentified phospholipid, three unidentified glycolipids and one unidentified lipid. The isoprenoid quinone was menaquinone (MK-10(H4)). The major fatty acids were anteiso-C15 : 0, anteiso-C15 : 1 A, C16 : 0, iso-C16 : 0, anteiso-C17 : 0 and iso-C17 : 0. The whole-cell sugars of strain THG-T121T were rhamnose, ribose, mannose and glucose. The peptidoglycan type of strain THG-T121T is A4β, containing l-Orn-D-Ser-L-Asp. The DNA G+C content of strain THG-T121T was 72.4 mol%. DNA-DNA hybridization values between strain THG-T121T and A. ferrariae KCTC 29134T, A. fermentans KCTC 3251T and C. carbonis KCTC 19824T were 30.2 % (27.3 %, reciprocal analysis), 28.4 %, (17.3 %) and 16.9 %, (9.3 %), respectively. On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain THG-T121T represents a novel species of the genus Actinotalea, for which the name Actinotaleasolisilvae sp. nov. is proposed. The type strain is THG-T121T (=KACC 19191T=CGMCC 4.7389T).

Corynebacterium defluvii sp. nov., isolated from Sewage

A Gram-positive, aerobic, non-motile, rod-shapeds, catalase-positive, and oxidase-negative strain, designated Y49T, was isolated from sewage collected from Jilin Agricultural University, China. It grew at 20–40°C (optimum at 30°C), at pH 6.0–8.0 (optimum at 7.0) and at 0–1.0% sodium chloride (optimum at 0%). The major isoprenoid quinone was menaquinone-8 (MK-8) and the polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmethylethanolamine, four unidentified lipids, and two unidentified aminolipids. The peptidoglycan was meso-diaminopimelic acid. The cell-wall sugars were galactose, arabinose, and glucose. The fatty acids were C9:0, C16:0, C16:1ω9c, C17:1ω9c, C18:3ω6c (6,9,12), C18:1ω9c, and C18:0. The DNA G+C content was 51.4 mol%. Based on the 16S rRNA gene sequence analysis, the nearest phylogenetic neighbors of strain Y49T were Corynebacterium efficiens DSM 44549T (97.5%), Corynebacterium callunae DSM 20147T (97.2%), Corynebacterium deserti GIMN 1.010T (96.8%), Corynebacterium glutamicum ATCC 13032T (96.4%), and other species belonging to this genus (92.3–95.4%). The DNA-DNA relatedness value between strain Y49T and C. efficiens DSM 44549T, C. callunae DSM 20147T, C. deserti GIMN1.010T, and C. glutamicum ATCC 13032T was 25.5±2.0%, 21.1±1.0%, 16.5±0.5%, and 13.5±0.9%, respectively. Based on the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain Y49T represents a novel species of the genus Corynebacterium, for which the name Corynebacterium defluvii sp nov. is proposed. The type strain is Y49T (= KCTC 39731T =CGMCC 1.15506T).

Altererythrobacter deserti sp. nov., isolated from desert soil

A Gram-stain-negative, aerobic, short rod-shaped, non-motile bacterium (THG-S3T), was isolated from desert soil. Growth occurred at 15-35 °C (optimum 28 °C), at pH 5-10 (optimum 7) and at 0-4 % NaCl (optimum 1 %). Based on 16S rRNA gene sequence analysis, the nearest phylogenetic neighbours of strain THG-S3T were identified as Altererythrobacter rigui KCTC 42620T (99.0 %), Altererythrobacter dongtanensis KCTC 22672T (97.1 %), Altererythrobacter xinjiangensis CCTCC AB 207166T (96.9 %), Altererythrobacter troitsensis KCTC 12303T (96.9 %). Levels of relatedness among strain THG-S3T and other Altererythrobacter species were lower than 96.0 %. DNA-DNA hybridization values between strain THG-S3T and A. rigui KCTC 42620T, A. dongtanensis KCTC 22672T, A. xinjiangensis CCTCC AB 207166T and A. troitsensis KCTC 12303T were 59.7 % (42.8 %, reciprocal analysis), 45.1 % (36.3 %), 34.7 % (25.1 %) and 15.1 % (12.3 %), respectively. The DNA G+C content of strain THG-S3T was 69 mol%. The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and three unidentified lipids The quinone was ubiquinone-10. The major fatty acids were C16 : 0, C17 : 1 ω6c, C18 : 1 ω7c and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c). On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain THG-S3T represents a novel species of the genus Altererythrobacter, for which the name Altererythrobacter deserti sp. nov. is proposed. The type strain is THG-S3T (=KACC 19190T=CGMCC 1.15959T).

Chryseomicrobium deserti sp. nov., isolated from desert soil in South Korea

A Gram-stain positive, aerobic, non-motile, rod-shaped bacterium (THG-T1.18T) was isolated from desert soil. Growth occurred at 20-35 °C (optimum 28-30 °C), at pH 5-7 (optimum 7) and at 0-4 % NaCl (optimum 0-1 %). Based on 16S rRNA sequence analysis, the nearest phylogenetic neighbours of strain THG-T1.18T were identified as Chryseomicrobium amylolyticum DSM 23442T (96.6 %), Chryseomicrobium imtechense JCM 16573T (96.3 %) and Chryseomicrobium aureum KACC 17219T (96.1 %). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unidentified aminolipids and one unidentified glycolipid. The quinone system was composed of MK-7, MK-8 and MK-6. The major fatty acids were iso C15 : 0 and anteiso C15 : 0. The type of peptidoglycan was A4β, containing of l-Orn-D-Glu. The DNA G+C content of strain THG-T1.18T was 50.4 mol%. DNA-DNA hybridization values between strain THG-T1.18T and C. amylolyticum DSM 23442T, C. imtechense JCM 16573T, C. aureum KACC 17219T were 24.7 % (20.1 % reciprocal analysis), 19.5 % (16.1 %) and 10.4 % (6.7 %) respectively. On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain THG-T1.18T represents a novel species of the genus Chryseomicrobium, for which the name Chryseomicrobium deserti sp. nov. is proposed. The type strain is THG-T1.18T (=KACC 18929T=CCTCC AB 2016179T).

Niastellahibisci sp. nov., isolated from rhizosphere soil of mugunghwa, the Korean national flower.

A Gram-stain-negative, aerobic, non-motile, long rods with no flagellum strain, designated THG-YS3.2.1T, was isolated from rhizosphere soil of mugunghwa, collected from Kyung Hee University, Yongin, South Korea. Growth occurred at 10-40 °C (optimum 28 °C), at pH 6.0-8.0 (optimum pH 7.0) and with 0-1.0 % NaCl (optimum 1.0 %). The predominant menaquinone was menaquinone-7 (MK-7). The major cellular fatty acids were anteiso-C12 : 0, iso-C14 : 0, iso-C15 : 0, anteiso-C15 : 0, iso-C15 : 1 G, anteiso-C15 : 1 A, C15 : 0 2-OH, C16 : 0, iso-C16 : 0, C16 : 0 3-OH, iso-C16 : 0 3-OH, iso-C16 : 1 G, C17 : 0 2-OH, iso-C17 : 0 3-OH, C17 : 1ω6c, C18 : 3ω6c (6, 9, 12), C18 : 0, summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH). The major polar lipids were phosphatidylmethylethanolamine), phosphatidylethanolamine, five unidentified aminolipids and three unidentified lipids. The DNA G+C content of strain THG-YS3.2.1T was 45.3 mol%. Based on 16S rRNA gene sequence analysis, the nearest phylogenetic neighbours of strain THG-YS3.2.1T were Niastella populi KCTC 22560T (98.7 % 16S rRNA gene sequence similarity), Niastella gongjuensis KACC 17399T (96.9 %), Niastella vici KCTC 42474T (96.2 %), Niastella yeongjuensis KACC 11466T (95.5 %) and Niastella koreensis KACC 11465T (95.1 %). DNA-DNA hybridization values between strain THG-YS3.2.1T and N. populi KCTC 22560T, N. gongjuensis KACC 17399T, N.vici KCTC 42474T, N. yeongjuensis KACC 11466T and N. koreensis KACC 11465Twere 55.8±1.0, 39.5±0.5, 35.2±0.1, 17.6±0.3 and 12.5±1.2 %, respectively. On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain THG-YS3.2.1T represents a novel species of the genus Niastella, for which the name Niastellahibisci sp. nov. is proposed. The type strain is THG-YS3.2.1T (=KCTC 52084T=CCTCC AB 2015356T).

Active Compounds from Schisandra chinensis Exhibiting Tyrosinase Activity and Melanin Content Inhibition in B16 Melanoma Cells

Schisandra chinensis has been used as traditional medicine. The structures of isolate active compounds (schisandrin B, deoxyschisandrin, schisandrin C) from S. chinensis were characterized by physical and spectroscopic analyses. Active compounds were tested for their potential to act as anti-melanogenesis or skin-whitening agents by their abilities to inhibit tyrosinase activity in the cell-free mushroom tyrosinase assay and cellular tyrosinase derived from B16 melanoma cells. The tyrosinase inhibitory activity was correlated to the inhibition of melanin productions in a-MSH-stimulated and unstimulated B16 cells. Cellular tyrosinase kinetics were analyzed and showed by Lineweaver- Burk plot. Schisandrin B was minimally cytotoxic (cell viability: 88.99% at 0.75 µM) and the IC50 value for suppression of mushroom tyrosinase activity was estimated as 0.6 µM. Zymography analysis demonstrated schisandrin B’s concentration-dependent effects and the kinetic analysis indicated schisandrin B’s noncompetitive-inhibitory action.