Acinetobacter faecalis Sp. Nov., Isolated from Elephant Faeces.

A Gram-negative coccobacillus, YIM 103518T, isolated from wild elephant feces in Xishuangbanna, Yunnan Province, West China, was characterized and identified using a polyphasic taxonomic approach. The strain was strictly aerobic, non-motile, catalase-positive and oxidase-negative, colonies were round, convex, smooth, and pale yellow. The strain growth at 4-40 ℃ (optimum, 28 ℃), pH 6.0-10.0 (optimum, pH 7.0) and 0-4% NaCl (optimum, 0%) in culture medium YIM 38. The major fatty acids of strain YIM 103518T were summed feature 3 (C16:1 ω6c/C16:1 ω7c), C16:0, and C18:1 ω9c. The predominant ubiquinone was Q-9. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine and phospholipids. The 16S rRNA gene sequence showed moderate level of similarity with Acinetobacter portensis AC 877T (98.7%), Acinetobacter sichuanensis CCTCC AB 2018118T (97.1%), and Acinetobacter cumulans CCTCC AB 2018119T (97.1%). The G+C content of the genomic DNA was 36.5 mol%. Strain YIM 103518T showed an average nucleotide identity value of 86.6%, 77.3% and 78.5%, a digital DNA-DNA hybridizations value of 31.2%, 21.9% and 23.0% with the type strain of A. portensis, A. sichuanensis and A. cumulans based on draft genome sequences, respectively. The results of the phenotypic, chemotaxonomic and phylogenetic analyses, showed that strain YIM 103518T represents a novel species of the genus Acinetobacter, for which the name Acinetobacter faecalis sp. nov. is proposed. The type strain is YIM 103518T (=CCTCC AB 2019201T = NBRC 114057T).

Nakamurella leprariae sp. nov., isolated from a lichen sample.

A novel actinobacterium, YIM 132084T, was isolated from Lepraria sp. lichen collected from Yunnan province, south-west PR China and identified by a polyphasic taxonomic approach. The strain was Gram-stain-positive, aerobic, catalase-positive, oxidase-negative, non-motile and coccus-shaped. Colonies were round, convex, smooth and light orange yellow in color. It grew at 10-40 °C (optimum 28 °C), at pH 6.0-11.0 (optimum pH 7.0) and in the presence of 0-4% NaCl (optimum 0%). Strain YIM 132084T comprised diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol as the major polar lipids, MK-8(H4) as the predominant menaquinone, and anteiso-C15:0, anteiso-C17:0, iso-C15:0 and iso-C16:0 as major fatty acids. Strain YIM 132084T had meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, and mannose, ribose, glucose and rhamnose as whole-cell sugars. The 16S rRNA gene sequence showed high level of similarity with Nakamurella flavida KCTC 19127T (97.7%) and Nakamurella flava CGMCC 4.7524T (97.7%). The G + C content of the genomic DNA was 72.4 mol%. Based on draft genome sequences, strain YIM 132084T showed an average nucleotide identity value of 76.1% and 74.9%, a digital DNA-DNA hybridization value of 20.9% and 20.6% with the reference strains Nakamurella flavida and Nakamurella flava, respectively. The results of the phenotypic, chemotaxonomic and phylogenetic analyses showed that strain YIM 132084T represents a novel species of the genus Nakamurella, for which the name Nakamurella leprariae sp. nov. is proposed. The type strain is YIM 132084T (= CGMCC 4.7667T = NBRC 114280T = KCTC 49367T).

Paenibacillus oceani sp. nov., isolated from surface seawater.

A Gram-stain-positive and motile bacterial strain, designated IB182363T, was isolated from surface seawater of the South China Sea. Cells grew at pH 5.0-9.5 (optimum, pH 7.0-8.0), 20-40 °C (optimum, 30 °C) and with 1-8 % (w/v) NaCl (optimum, 2-4 %). On the basis of 16S rRNA gene sequence analysis, strain IB182363T was affiliated to the genus Paenibacillus and the closest phylogenetically related species was Paenibacillus ginsengarvi DSM18677T with 96.9 % sequence similarity. The values of whole genome average nucleotide identity analysis and digital DNA-DNA hybridization between the isolate and the closely related type strains were less than 86.3 and 25.6 %, respectively. Chemotaxonomic analysis revealed that strain IB182363T possessed meso-diaminopimelic acid in the cell-wall peptidoglycan and contained menaquinone MK-7 as the predominant isoprenoid quinone. The major cellular fatty acids were anteiso-C15 : 0, C16 : 0 and iso-C16 : 0. The polar lipids comprised phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, one unidentified glycolipid, two unidentified aminolipids, two unidentified phospholipids and four unidentified aminophospholipids. The genomic DNA G+C content was 54.5 mol%. On the basis of the above results, strain IB182363T represents a novel species of the genus Paenibacillus, for which we propose the name Paenibacillus oceani sp. nov. with the type strain IB182363T (=MCCC 1K04630T=JCM 34214T).

Nocardioides flavescens sp. nov., isolated from soil of Gaoligong Mountain, PR China.

A Gram-stain-positive, non-motile, aerobic, rod-shaped, golden yellow-pigmented actinobacterium, designated strain YIM 123512T, was isolated from soil sampled at Gaoligong Mountain, Yunnan Province, PR China. The strain grew at 10-40 °C (optimum, 28 °C), with 0-3.0 % (w/v) NaCl (optimum, 0-1.0 %) and at pH 5.0-11.0 (optimum, pH 7.0). The major menaquinone was MK-8(H4) and the major cellular fatty acids (>10 %) were identified as iso-C16 : 0 and C17 : 1 ω8c. Strain YIM 123512T had ll-2, 6-diaminopimelic acid as the diagnostic cell-wall diamino acid. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylglycerol, two unknown phospholipids and one unknown lipid. The genomic DNA G+C content of strain YIM 123512T was 72.7 mol% based on its draft genome sequence. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain YIM 123512T appeared to be most closely related to Nocardioides halotolerans KCTC 19274T (97.7 % sequence similarity) and to belong to the genus Nocardioides. The sequence similarity values of strain YIM 123512T to other currently described type strains of the genus Nocardioides were less than 97.0 %. Based on the draft genome sequence, the novel strain showed an average nucleotide identity value of 80.2 % and a digital DNA-DNA hybridization value of 23.1 % with the reference strain N. halotolerans KCTC 19274T. The results of the polyphasic taxonomic study including phenotypic, chemotaxonomic and phylogenetic analyses show that strain YIM 123512T represents a novel species of the genus Nocardioides, for which the name Nocardioides flavescens sp. nov. is proposed. The type strain is YIM 123512T (=KCTC 49303T=CGMCC 4.7628T).

Ottowia flava sp. nov., isolated from fish intestines.

A novel Gram-negative bacterium, non-motile and short rod-shaped, designated strain GY511T, was isolated from the intestines of fish collected from Maowei Sea, China. Growth occurred at pH 6.0-9.0 (optimum 7.0), 4-37 °C (optimum 28 °C) and at 0-2.5% (w/v) NaCl (optimum 1.0%). The result of 16S rRNA gene sequence analysis showed that strain GY511T is closely related to O. oryzae NBRC 113109T (97.6%), O. konkukae DSM 105395T (97.4%), Ottowia beijingensis CGMCC 1.12324T (95.9%), Ottowia pentelensis DSM 21699T (95.2%) and Ottowia thiooxydans DSM 14619T (95.0%). The DNA-DNA hybridization values of strain GY511T with O. oryzae NBRC 113109T and O. konkukae DSM 105395T were 35.4 ± 3.1% and 26.3 ± 1.8%, respectively. The major fatty acids (> 10%) were identified as summed feature 3 (C16:1ω7c and/or C16:1ω6c), C16:0 and summed feature 8 (C18:1ω7c and/or C18:1ω6c) and the major respiratory quinone was ubiquinone-8 (Q-8). The polar lipids comprised diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmethylethanolamine, two unidentified aminolipids and an unidentified phospholipid. The G+C content of the genomic DNA was 62.9 mol%. Thiosulfate could be utilized as co-substrate for aerobic growth and was oxidised to sulfate. On the basis of phenotypic, chemotaxonomic and molecular data, strain GY511T is considered to represent a novel species of the genus Ottowia, for which the name Ottowia flava sp. nov. is proposed. The type strain is GY511T (= NBRC 113500T = DSM 107425T = CGMCC 1.13650T).

Falsibacillus albus sp. nov., isolated from mangrove soil.

A novel Gram-stain-positive, catalase- and oxidase-positive, endospore-forming bacterium, designated GY 10110T, was isolated from mangrove soil collected from Qinzhou, Guangxi province, China. Cells were aerobic, motile with peritrichous flagella and rod-shaped. The strain grew at 15-37 °C (optimum, 28 °C), at 0-3 %(w/v) NaCl (1 %) and at pH 6.0-9.0 (pH 7.0). The major fatty acids of strain GY 10110T were anteiso-C15 : 0, iso-C15 : 0 and iso-C16 : 0. The predominant menaquinone was MK-7. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The polar lipid profile comprised diphosphatidylglycerol, phosphatidylethanolamine, phosphoglycolipid, glycolipid, two unidentified aminophospholipids and three unidentified phospholipids. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain GY 10110T was closely related to Falsibacillus pallidus CCTCC AB 207188T (98.0 % sequence similarity) and Bacillus oceanisediminis CGMCC 1.10115T (96.9 %), respectively. The G+C content of strain GY 10110T based on the whole genome sequence was 42.3 mol%. The novel strain showed an average nucleotide identity (ANI) value of 77.8 % and a digital DNA-DNA hybridization (dDDH) value of 15.6 % with Falsibacillus pallidus CCTCC AB 207188T based on draft genome sequences, followed by Bacillus oceanisediminis CGMCC 1.10115T with ANI and dDDH values of 75.2 and 12.8 %, respectively. The results of the polyphasic taxonomic study, including phenotypic, chemotaxonomic and phylogenetic analysis, showed that strain GY 10110T represents a novel species of the genus Falsibacillus, for which the name Falsibacillus albus sp. nov. is proposed. The type strain is GY 10110T (=CGMCC 1.13648T=NBRC 113502T).

Marmoricola endophyticus sp. nov., an endophytic actinobacterium isolated from Thespesia populnea.

A novel endophytic actinobacterium, designated strain 8BXZ-J1T, was isolated from surface-sterilized branches of Thespesia populnea collected from Beilun Estuary Mangrove Forest National Nature Reserve in Guangxi, China, and examined by a polyphasic approach to determine its taxonomic position. Cells of the isolate were Gram-stain-positive, aerobic, non-spore-forming, non-motile and short rod-shaped. Phylogenetic analysis based on 16S rRNA gene sequences suggested that strain 8BXZ-J1T belonged to the genus Marmoricola, sharing highest similarity with Marmoricola solisilvae DSM 27140T (96.9 %). The isolate grew at 10-35 °C (optimum, 28-30 °C), at pH 6.0-8.0 (optimum, pH 7.0) and in the presence of 0-10 % (w/v) NaCl (optimum, 0-5.0 %). The organism contained ll-2,6-diaminopimelic acid as the diagnostic diamino acid of the peptidoglycan, MK-8(H4) as the major menaquinone, and a polar lipid profile including diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylinositol and two unknown lipids. The major fatty acids of strain 8BXZ-J1T were C18 : 0 10-methyl, iso-C16 : 0 and C16 : 0. The G+C content of the genomic DNA was 68.7 mol%. These data demonstrate that strain 8BXZ-J1T is representative of a novel species of the genus Marmoricola, for which the name Marmoricola endophyticus sp. nov. is proposed. The type strain is 8BXZ-J1T (=KCTC 39789T=CGMCC 1.16067T).

Microlunatus endophyticus sp. nov., an endophytic actinobacterium isolated from bark of Bruguiera sexangula.

A Gram-stain-positive, aerobic, coccoid, non-motile, non-spore-forming bacterium, designated strain S3Af-1T, was isolated from surface-sterilized bark of Bruguiera sexangula collected from Dongzhaigang National Nature Reserve in Hainan, China, and examined using a polyphasic approach to clarify its taxonomic position. This bacterium did not produce substrate mycelia or aerial hyphae, and no diffusible pigments were observed on the media tested. Strain S3Af-1T grew optimally without NaCl, at 28-30 °C and at pH 7.0.Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain S3Af-1T belonged to the genus Microlunatus and shared highest similarity with 'Microlunatus terrae' BS6 (97.43 %) and Microlunatus soli CC-12602T (97.08 %). DNA-DNA hybridization results indicated that the level of relatedness between strain S3Af-1T and M. soli CC-12602T was less than 70 %. The DNA G+C content of strain S3Af-1T was 67.1 mol%. The cell-wall peptidoglycan contained ll-2,6-diaminopimelic acid. MK-9(H6) and MK-9(H4) were the predominant menaquinones. Phosphatidylglycerol, diphosphatidylglycerol, an unidentified glycolipid, two unidentified phospholipids and other lipids were detected in the polar lipid extracts. The major fatty acids were iso-C16 : 0, anteiso-C17 : 0 and anteiso-C15 : 0. On the basis of phylogenetic analysis, and phenotypic and chemotaxonomic characteristics, strain S3Af-1T represents a novel species of the genus Microlunatus, for which the name Microlunatus endophyticus sp. nov. is proposed. The type strain is S3Af-1T ( = DSM 100019T = CGMCC 4.7306T).

Isolation and characterization of a new bioactive isoflavone from Derris eriocarpa.

Derris eriocarpa How is an important medicinal plant, which is used as Zhuang ethnomedicine and Dai ethnomedicine to treat various diseases. One new compound, 3',4'-di-O-methylene-5-hydroxy-7-methoxy-6-isopentenyl isoflavone (1) and a known synthetic but new naturally occurring compound trans-3,4,5-trimethoxy-4'-isopentenyloxyl-stilbene (2), together with five known compounds, 5,7-dimethoxy-6-(3-methyl-2-butenyl)-4'-hydroxyl isoflavones (3), robustone (4), trans-3,4,5,4'-tetramethoxy-stilbene (5), robustic acid (6), and robustin (7) were isolated from the stem of D. eriocarpa. Spectroscopic analysis revealed the chemical structures of compounds 1-7.. Compounds 1 and 3 exhibited significant scavenging activities against 1,1-diphenyl-2-picrylhydrazyl radical and superoxide anions. Compounds 1-3 exhibited potent antiproliferative activity on Hela cells.

Synthesis, biological evaluation and 3D-QSAR study of novel 4,5-dihydro-1H-pyrazole thiazole derivatives as BRAF(V6°°E) inhibitors.

Many reports implied that the BRAF serine/threonine kinase was mutated in various types of human tumors, which were related with cell growth, survival and differentiation. To provide new therapeutic opportunities, a series of novel 4,5-dihydro-1H-pyrazole derivatives (6a-10d) containing thiazole moiety as potential V600E mutant BRAF kinase (BRAF(V600E)) inhibitors were designed and synthesized. All compounds were evaluated in vitro for anticancer activities against WM266.4 human melanoma cell line and breast cancer MCF-7 cell line. Compound 10d displayed the most potential antiproliferative activity with an IC50 value of 0.12µM against cell line WM266.4 and 0.16µM against MCF-7 with positive control Sorafenib. Results of the inhibitory activity against BRAF(V600E) revealed that compound 10d was bearing the best bioactivity with IC50 of 0.05µM as well. On the basis of the result of flow cytometry, with the dose of compound 10d increasing, more and more cancer cell gradually encountered apoptosis or died, which indicated the compound 10d could induce remarkable apoptosis of MCF-7 and WM266.4 cells in a dose dependent manner. Furthermore, docking simulation of inhibitor analogues and 3D-QSAR modeling provided potential binding model and further knowledge of pharmacophore.