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Conceptualization, T.-H.Y., S.Z. and S.O.; methodology, S.Z. and M.F; software, S.O., S.Z. and T.T.M.N.; validation, S.O., M.F. and S.Z.; formal analysis, S.Z., S.O. and M.F.; investigation, T.-H.Y., S.O. and J.J. (Jeehaeng Jeong); resources, A.D.B., J.J. (Jeyong Jung) and T.T.M.N.; data curation, S.Z., J.J. (Jeyong Jung) and M.F.; writing—S.O. and A.D.B.; writing—review and editing, T.-H.Y. and S.O.; visualization, S.Z., A.D.B., T.T.M.N. and J.J. (Jeyong Jung); supervision, T.-H.Y. and J.J. (Jeehaeng Jeong); project administration, T.-H.Y. and J.J. (Jeehaeng Jeong). All authors have read and agreed to the published version of the manuscript.
Figure 1. Phylogenetic tree of S. chattanoogensis THA-663. Bootstrap percentages (1000 replicates) above 70% are shown at the nodes and toggle-scaled the tree.
Figure 1. Phylogenetic tree of S. chattanoogensis THA-663. Bootstrap percentages (1000 replicates) above 70% are shown at the nodes and toggle-scaled the tree.
Figure 2. DPPH and ABTS radical scavenging activity of THA-663S. DPPH radical (A) and ABTS+ cation (B) scavenging activity of THA-663S. The radical scavenging effect was presented as a percentage measured in the control group. Ascorbic acid serves as a positive control. IC50 values measured for ascorbic acid scavenging activity on DPPH and ABTS radicals were 521.2 and 160.5 ppm, respectively. The results are shown as the mean ± SD of three independent experiments.
Figure 2. DPPH and ABTS radical scavenging activity of THA-663S. DPPH radical (A) and ABTS+ cation (B) scavenging activity of THA-663S. The radical scavenging effect was presented as a percentage measured in the control group. Ascorbic acid serves as a positive control. IC50 values measured for ascorbic acid scavenging activity on DPPH and ABTS radicals were 521.2 and 160.5 ppm, respectively. The results are shown as the mean ± SD of three independent experiments.
Figure 3. Cell viability and intracellular reactive oxygen species (ROS) generations in UVB-exposed HaCaT cells treated with THA-663S. Following UVB irradiation (125 mJ/cm2), HaCaT cells were THA-663. S (0.1, 1, and 10 µg/mL) for 24 h. ROS levels were determined using flow cytometry (A). The relative intensity of ROS generation is shown in the bar graph (B). Cell viability was measured using an MTT assay (C). The results were shown as the mean ± SD of three independent experiments. ### p < 0.001 compared with non-irradiated group, * p < 0.05, ** p < 0.01, *** p < 0.001 compared with only UVB-irradiated group.
Figure 3. Cell viability and intracellular reactive oxygen species (ROS) generations in UVB-exposed HaCaT cells treated with THA-663S. Following UVB irradiation (125 mJ/cm2), HaCaT cells were THA-663. S (0.1, 1, and 10 µg/mL) for 24 h. ROS levels were determined using flow cytometry (A). The relative intensity of ROS generation is shown in the bar graph (B). Cell viability was measured using an MTT assay (C). The results were shown as the mean ± SD of three independent experiments. ### p < 0.001 compared with non-irradiated group, * p < 0.05, ** p < 0.01, *** p < 0.001 compared with only UVB-irradiated group.
Figure 4. THA-663S inhibited the secretion of MMP-1 and MMP-3 in UVB-irradiated HaCaT cells. Following UVB irradiation (125 mJ/cm2), HaCaT cells were treated with or without the indicated concentration of THA-663S (0.1, 1, and 10 µg/mL) for 24 h. MMP-1 (A) and MMP-3 (B) levels were determined using ELISA kits. The results are shown as the mean ± SD of three independent experiments. # p < 0.05, ### p < 0.001 compared with non-irradiated group, * p < 0.05 compared with only UVB-irradiated group.
Figure 4. THA-663S inhibited the secretion of MMP-1 and MMP-3 in UVB-irradiated HaCaT cells. Following UVB irradiation (125 mJ/cm2), HaCaT cells were treated with or without the indicated concentration of THA-663S (0.1, 1, and 10 µg/mL) for 24 h. MMP-1 (A) and MMP-3 (B) levels were determined using ELISA kits. The results are shown as the mean ± SD of three independent experiments. # p < 0.05, ### p < 0.001 compared with non-irradiated group, * p < 0.05 compared with only UVB-irradiated group.
Figure 5. MMP-1, TGF-β1, and procollagen type I mRNA expression on HaCaT cells of THA-663S. Following UVB irradiation (125 mJ/cm2), HaCaT cells were treated with or without the shown concentrations of THA-663S (0.1 and 1 µg/mL) for 24 h. The MMP-1, TGF-β1, and procollagen type I mRNA levels were measured using RT-PCR analysis (A). The band intensities were quantified by densitometry, normalized to the level of GAPDH, and calculated as the percentage of the non-irradiated group (B). The results are shown as the mean ± SD of three independent experiments. # p < 0.05, ## p < 0.01 compared with non-irradiated group, * p < 0.05, ** p < 0.01, *** p < 0.001 compared with only UVB-irradiated group.
Figure 5. MMP-1, TGF-β1, and procollagen type I mRNA expression on HaCaT cells of THA-663S. Following UVB irradiation (125 mJ/cm2), HaCaT cells were treated with or without the shown concentrations of THA-663S (0.1 and 1 µg/mL) for 24 h. The MMP-1, TGF-β1, and procollagen type I mRNA levels were measured using RT-PCR analysis (A). The band intensities were quantified by densitometry, normalized to the level of GAPDH, and calculated as the percentage of the non-irradiated group (B). The results are shown as the mean ± SD of three independent experiments. # p < 0.05, ## p < 0.01 compared with non-irradiated group, * p < 0.05, ** p < 0.01, *** p < 0.001 compared with only UVB-irradiated group.
Figure 6. MAPK and AP-1 signaling protein expression in HaCaT cells of THA-663S. Following UVB irradiation (125 mJ/cm2), HaCaT cells were treated with or without the indicated concentration of THA-663S (0.1, 1, and 10 µg/mL) for 1.5 h and 4 h. The p-p38, p-ERK, and p-JNK protein levels were measured using Western blotting (A). The band intensities were quantified by densitometry, normalized to the level of β-actin, and calculated as the percentage of the non-irradiated group (B). The p-c-jun and p-c-fos protein levels were measured using Western blot (C). The band intensities were quantified by densitometry, normalized to the level of β-actin, and calculated as the percentage of the non-irradiated group (D). The results are shown as the mean ± SD of three independent experiments. # p < 0.05, ## p < 0.01, ### p < 0.001 compared with non-irradiated group, * p < 0.05, ** p < 0.01, *** p < 0.001 compared with only UVB-irradiated group.
Figure 6. MAPK and AP-1 signaling protein expression in HaCaT cells of THA-663S. Following UVB irradiation (125 mJ/cm2), HaCaT cells were treated with or without the indicated concentration of THA-663S (0.1, 1, and 10 µg/mL) for 1.5 h and 4 h. The p-p38, p-ERK, and p-JNK protein levels were measured using Western blotting (A). The band intensities were quantified by densitometry, normalized to the level of β-actin, and calculated as the percentage of the non-irradiated group (B). The p-c-jun and p-c-fos protein levels were measured using Western blot (C). The band intensities were quantified by densitometry, normalized to the level of β-actin, and calculated as the percentage of the non-irradiated group (D). The results are shown as the mean ± SD of three independent experiments. # p < 0.05, ## p < 0.01, ### p < 0.001 compared with non-irradiated group, * p < 0.05, ** p < 0.01, *** p < 0.001 compared with only UVB-irradiated group.
Figure 7. TGF-β/Smad signaling protein expression in HaCaT cells of THA-663S. Following UVB irradiation (125 mJ/cm2), HaCaT cells were treated with or without the indicated concentration of THA-663S (0.1, 1, and 10 µg/mL) for 1.5 h. The TGF-β1 and Smad7 protein levels were measured using Western blotting (A). The band intensities were quantified using densitometry, normalized to the level of β-actin, and calculated as the percentage of the non-irradiated group (B). The results are shown as the mean ± SD of three independent experiments. ### p < 0.001 compared with non-irradiated group, * p < 0.05, ** p < 0.01, *** p < 0.001 compared with only UVB-irradiated group.
Figure 7. TGF-β/Smad signaling protein expression in HaCaT cells of THA-663S. Following UVB irradiation (125 mJ/cm2), HaCaT cells were treated with or without the indicated concentration of THA-663S (0.1, 1, and 10 µg/mL) for 1.5 h. The TGF-β1 and Smad7 protein levels were measured using Western blotting (A). The band intensities were quantified using densitometry, normalized to the level of β-actin, and calculated as the percentage of the non-irradiated group (B). The results are shown as the mean ± SD of three independent experiments. ### p < 0.001 compared with non-irradiated group, * p < 0.05, ** p < 0.01, *** p < 0.001 compared with only UVB-irradiated group.
Table 1. Differences in pH, temperature and NaCl concentration.
Table 1. Differences in pH, temperature and NaCl concentration.
CharacteristicTHA-663KACC 14678 2KACC 21704 3pH range 5–95–95–9Optimum pH 6–76–76–810 °C-W 1-30 °C+++37 °C+++Optimum temperature 303037Maximum NaCl con. (%, w/v)4%6%4%Table 2. Minimum inhibitory concentration (MIC) for antibiotics.
Table 2. Minimum inhibitory concentration (MIC) for antibiotics.
AntibioticsTHA-663KACC 14678 1KACC 21074 2Kanamycin1.01.01.5Ampicillin>256>256-1)Erythromycin1.50.380.38Streptomycin0.50.50.75Vancomycin0.380.380.38Clindamycin481.50.25Gentamycin0.380.250.38Tetracycline48832Chloramphenicol->256-Table 3. Antibacterial effect of THA-663S on MRSA.
Table 3. Antibacterial effect of THA-663S on MRSA.
MicroorganismsCollection Code No.THA-663SAMP 1NOR 2GEN 3NAT 4S. aureus 285CCARM 02041.31 ± 0.4%--1.1 ± 0.1%-S. aureus 503CCARM 02051.36 ± 0.3%--1.02 ± 0.2%-S. aureusCCARM 38551.02 ± 0.4%1.39 ± 0.1%---S. aureusCCARM 30891.01 ± 0.2%----Table 4. List of components in THA-663S identified by UPLC-Qtof-MS.
Table 4. List of components in THA-663S identified by UPLC-Qtof-MS.
No.Component NameObserved RT (min)FormulaObserved m/zMass Error (mDa)Adducts13,6,9-Trimethyl-benzo[de]chromene1.29 C15H14O233.0940 0.3 +Na22-(4-ethoxybenzylidene)malonic acid2.25 C11H10O5223.0598 −0.3 +H3N-[[3-[3-fluoro-4-(3-methyl-1,2,4-oxadiazol-5-yl)phenyl]-2-oxo-1,3-oxazolidin-5-yl]methyl]acetamide3.58 C16H18N2O6335.1226 −1.2 +H41H-Indole, 3-(1E)-1-propen-1-yl-3.80 C11H11N158.0964 0.0 +H5Kenganthranol A4.17 C25H28O5431.1810 −1.9 +Na6[(2-Methoxyphenyl)Table 5. MIC and MBC of THA-663S to multi-drug-resistant bacteria.
Table 5. MIC and MBC of THA-663S to multi-drug-resistant bacteria.
GramMicroorganismsCollection Code No.MICMBCGram-positiveS. aureus 285CCARM 02042.55S. aureus 503CCARM 02052.55S. aureusCCARM 38551.255S. aureusCCARM 308955
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