Cheng-Liang Cao

Nocardia endophytica sp. nov., an endophytic actinomycete isolated from the oil-seed plant Jatropha curcas L.

A novel actinomycete, designated strain KLBMP 1256(T), was isolated from a surface-sterilized stem of the oil-seed plant Jatropha curcas L. collected from Sichuan Province, south-west China, and was characterized to determine its taxonomic position. Phylogenetic analyses based on 16S rRNA gene sequences indicated that the isolate was closely related to members of the genus Nocardia in the family Nocardiaceae, being most closely related to Nocardia callitridis CAP 290(T) (98.4 % similarity) and Nocardia nova JCM 6044(T) (97.5 %). Levels of 16S rRNA gene sequence similarity between strain KLBMP 1256(T) and the type strains of other recognized species of the genus Nocardia were less than 97 %. Chemotaxonomic data supported the affiliation of the new isolate to the genus Nocardia. However, the novel strain could be distinguished from its closest phylogenetic neighbour, N. callitridis CAP 290(T), by a range of phenotypic properties. The combination of low DNA-DNA relatedness values and phenotypic differences from N. callitridis CAP 290(T) indicated that strain KLBMP 1256(T) represents a novel species of the genus Nocardia, for which the name Nocardia endophytica sp. nov. is proposed. The type strain is KLBMP 1256(T) ( = KCTC 19777(T)  = CCTCC AA 2010004(T)).

Schiff base – Chitosan grafted l-monoguluronic acid as a novel solid-phase adsorbent for removal of congo red

A novel modified chitosan adsorbent (GL-SBCS) was synthesized by covalently grafting a Schiff base-chitosan (SBCS) onto the surface of l-monoguluronic acid. Physico-chemical investigation on the adsorption of congo red, an anionic azo dye by GL-SBCS has been carried out. The effect of different weight contents of chitosan in GL-SBCS composite, adsorbent dosage, initial pH and contract time were studied in detail using batch adsorption. Results showed that GL-SBCS exhibited better than normal CS and l-monoguluronic acid. Further investigation demonstrated that the adsorption pattern fitted well with the Langmuir model (R(2)>0.99) but less-satisfied the Freundlich model. Both ionic interaction as well as physical forces is responsible for binding of congo red with GL-SBCS as determined by zeta potential measurement Both sodium chloride and sodium dodecyl sulfate significantly influenced the adsorption process. SBCS would be a good method and resource to increase absorption efficiency for the removal of anionic dyes in a wastewater treatment process.

Modestobacter roseus sp. nov., an endophytic actinomycete isolated from the coastal halophyte Salicornia europaea Linn., and emended description of the genus Modestobacter.

A novel actinomycete, designated strain KLBMP 1279(T), was isolated from surface-sterilized roots of a coastal halophyte, Salicornia europaea Linn., collected from Jiangsu Province, in the east of China. The taxonomic status of this organism was established using a polyphasic approach. 16S rRNA gene sequence analysis indicated that strain KLBMP 1279(T) was closely related to Modestobacter marinus 42H12-1(T) (99.5% 16S rRNA gene sequence similarity), Modestobacter versicolor CP153-2(T) (98.4%) and Modestobacter multiseptatus AA-826(T) (97.5%). Chemotaxonomic characteristics were consistent with its assignment to the genus Modestobacter in that the isolate had meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall, MK-9(H4) as major menaquinone and a polar lipid profile containing diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannosides, two unknown aminophospholipids and an unknown phospholipid. The predominant fatty acids were iso-C16:0, iso-C15:0 and C17:1ω8c. The DNA G+C content was 71.7 mol%. However, DNA-DNA hybridization assays as well as physiological and biochemical analyses differentiated strain KLBMP 1279(T) from its closest phylogenetic relatives. On the basis of phenotypic, chemotaxonomic and phylogenetic evidence, the isolate KLBMP 1279(T) represents a novel species of the genus Modestobacter, for which the name Modestobacter roseus sp. nov. is proposed; the type strain is KLBMP 1279(T) (=KCTC 19887(T)=NBRC 108673(T)=DSM 45764(T)). An emended description of the genus Modestobacter is also proposed.

Glycomyces phytohabitans sp. nov., a novel endophytic actinomycete isolated from the coastal halophyte in Jiangsu, East China

A novel endophytic actinomycete, designated strain KLBMP 1483T, was isolated from the stem of the coastal plant Dendranthema indicum (Linn.) Des Moul collected from Nantong, in East China. Phylogenetic analysis showed that strain KLBMP 1483T was affiliated with the genus Glycomyces within the family Glycomycetaceae and shared the highest 16S rRNA gene sequence similarities with the type strains of Glycomyces arizonensis NRRL B-16153T (96.7%) and Glycomyces tenuis IFO 15904T (96.2%), and lower similarities (94.1–95.1%) to the other members of the genus Glycomyces, which distinguished KLBMP 1483T from representatives of the genus Glycomyces. The whole-cell hydrolysates contained meso-diaminopimelic acid, glucose, xylose and galactose. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides, two unknown aminophospholipids, two phosphoglycolipids, two unknown phospholipids and one unknown lipid. MK-10(H4) was the predominant menaquinone. The major fatty acids were iso-C15:0, anteiso-C15:0, iso-C16:0, iso-C16:1 G and anteiso-C17:0. On the basis of the phenotypic and genotypic characteristics presented in this study, strain KLBMP 1483T represents a novel species, for which the name Glycomyces phytohabitans sp. nov. is proposed. The type strain is KLBMP 1483T (NBRC 109116T=DSM 45766T).

Streptomyces halophytocola sp. nov., an endophytic actinomycete isolated from the surface-sterilized stems of a coastal halophyte Tamarix chinensis Lour.

A novel actinomycete, designated KLBMP 1284(T), was isolated from the surface-sterilized stems of a coastal halophyte Tamarix chinensis Lour. collected from the city of Nantong, Jiangsu Province, east China. The strain was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. Analysis of the 16S rRNA gene sequence of strain KLBMP 1284(T) revealed that the strain formed a distinct clade within the phylogenetic tree based on 16S rRNA gene sequences and the highest sequence similarity (99.43 %) was to Streptomyces sulphureus NRRL B-1627(T). 16S rRNA gene sequence similarity to other species of the genus Streptomyces was lower than 97 %. Based on DNA-DNA hybridization values and comparison of morphological and phenotypic data, KLBMP 1284(T) could be distinguished from the closest phylogenetically related species, Streptomyces sulphureus NRRL B-1627(T). Thus, based on these data, it is evident that strain KLBMP 1284(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces halophytocola sp. nov. is proposed. The type strain is KLBMP 1284(T) (= KCTC 19890(T) = NBRC 108770(T)).

Nocardiajiangsuensis sp. nov., an actinomycete isolated from coastal soil

A novel actinomycete strain, designated strain KLBMP S0027T, was isolated from a coastal soil collected from the coastal region of Lianyungang, Jiangsu Province, in east China, and was studied in detail for its taxonomic position. Comparative 16S rRNA gene sequence analysis showed that this strain belonged to the genus Nocardia and was most closely related to Nocardia harenae WS-26T (98.5 %), Nocardiaasiatica NBRC 100129T (98.5 %), Nocardiaabscessus NBRC 100374T (98.2 %), Nocardia brasiliensis NBRC 14402T (98.2 %) and Nocardiacyriacigeorgica NBRC 100375T (98 %), respectively. The strain showed a combination of chemotaxonomic and morphological properties typical of the genus Nocardia. The cell wall contained meso-diaminopimelic acid (type IV), and whole-cell sugars were arabinose, galactose, glucose and ribose. Strain KLBMP S0027T contained MK-8(H4ω-cycl) as the predominant menaquinone; C16 : 0, C18 : 1ω9c, C18 : 0 10-methyl (TBSA) and summed feature 3 (C16 : 1ω7c/C16 : 1ω6c) were the major cellular fatty acids. The polar lipids detected were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, one unknown glycolipid and two unknown lipids. Mycolic acids were detected. The G+C content of the DNA was 70.5 %. However, a combination of DNA-DNA hybridization and phenotypic data demonstrated that strain KLBMP S0027T could be clearly distinguished from the type strain of the most closely related species, N. harenae WS-26T. On the basis of the data presented from a polyphasic study, it was evident that this strain should be assigned to a novel species of the genus Nocardia, for which the name Nocardia jiangsuensis sp. nov. is proposed. The type strain is KLBMP S0027T (=CGMCC 4.7330T=KCTC 39691T).

Methylated actinomycin D, a novel actinomycin D analog induces apoptosis in HepG2 cells through Fas- and mitochondria-mediated pathways.

Actinomycin D (Act D), a well known of clinical antitumor drug, has been used for the treatment of some highly malignant tumors, however, the clinical application was limited by its extreme cytotoxicity. In the present study, we reported that methylated actinomycin D (mAct D), a novel actinomycin D analog isolated from Streptomyces sp. KLBMP 2541 in our previous study, could not only exert stronger inhibitory effects on several human cancer cells than Act D in dose‐ and time‐dependent manner at ng concentrations, especially on HepG2 cells, but also lower cytotoxicity in normal cells (HL‐7702). Base on these results, HepG2 cells were treated for further study to illustrate the potential mechanism of mAct D. The results of nuclei morphology examination, DNA fragmentation detection, sub‐G1 analysis, annexin V‐FITC/PI staining and activation of caspase‐3 indicated mAct D significantly induced HepG2 cells apoptosis. Semiquantitative RT‐PCR and Western blot analysis revealed that mAct D induced apoptosis in HepG2 cells through mitochondria‐dependent pathway by increasing levels of caspase‐9, Bax, Bak while decreasing levels of Bcl‐2, Bid, and Fas‐dependent pathway by increasing levels of Fas, FasL, FADD, and caspase‐8. Subsequently, pretreatment with specific inhibitor of caspase‐8 Z‐LEHD‐FMK and caspase‐9 Z‐LEHD‐FMK significantly attenuated caspase‐3 activity, the cleavage of caspase‐3 and PARP, meanwhile increased the cell viability. In addition, p53 and mitochondrial transcription factor A (mtTFA) were also upregulated. Taken together, ng concentrations mAct D induces the apoptosis of HepG2 through Fas‐ and mitochondria‐mediated pathway and presents a potential novel alternative agent for the treatment of human hepatic carcinoma.

Nocardia rhizosphaerihabitans sp. nov., a novel actinomycete isolated from a coastal soil

An actinomycete strain, designated KLBMP S0039T, was isolated from the rhizosphere soil of Lycium Linn., collected from the coastal region in Lianyungang, Jiangsu Province, eastern PR China, and was studied to determine its taxonomic position. The isolate showed a combination of morphological and chemotaxonomic properties typical of the members of the genus Nocardia. The cell wall contained meso-diaminopimelic acid as the diagnostic diamino acid and the whole-cell sugars were galactose, arabinose, glucose and ribose. The predominant menaquinone was identified as MK-8(H4ω-cycl). The diagnostic phospholipids were found to be diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and unknown lipids. The predominant cellular fatty acids were identified as C16 : 0, C18 : 0, C18 : 1ω9c, and 10-methyl C18 : 0 [tuberculostearic acid (TBSA)]. The G+C content of the genomic DNA was determined to be 68.2 mol%. The 16S rRNA gene sequence similarity indicated that KLBMP S0039T was most closely related to Nocardia neocaledoniensis NBRC 108232T (99.4 % 16S rRNA gene sequence similarity) and Nocardia asteroides NBRC 15531T (99.2 %), similarities to other type strains of species of the genus Nocardia were found to be less than 98.6 %. However, DNA-DNA relatedness values and phenotypic data indicated that KLBMP S0039T could be clearly distinguished from the closely related species of the genus Nocardia. On the basis of polyphasic taxonomic data, it is concluded that KLBMP S0039T represents a novel species of the genus Nocardia, for which the name Nocardiarhizosphaerihabitans sp. nov. is proposed. The type strain is KLBMP S0039T (=KCTC 39693T=CGMCC 4.7329T).

Actinophytocola glycyrrhizae sp. nov. isolated from the rhizosphere of Glycyrrhiza inflata

A Gram-stain-positive, aerobic actinomycete, designated strain BMP B8152T, was isolated from the rhizosphere of Glycyrrhiza inflata collected ashore, in Kashi, Xinjiang province, northwest PR China. A polyphasic approach was used to establish the taxonomic position of this strain. BMP B8152T was observed to form non-fragmented substrate mycelium, and relatively scanty aerial mycelium with rod-shaped spores. Cell-wall hydrolysates contained meso-diaminopimelic acid, galactose, arabinose, glucose and rhamnose (trace). Mycolic acids were not detected. The diagnostic phospholipids were identified as diphosphatidylglycerol, phosphatidylethanolamine, hydroxyphosphatidylethanolamine, ninhydrin-positive phosphoglycolipid and phosphatidylinositol. The predominant menaquinone and fatty acid were MK-9(H4) and iso-branched hexadecanoate (iso-C16 : 0), respectively. The phylogenetic analyses based on the 16S rRNA gene sequences indicated that BMP B8152T formed a distinct monophyletic clade clustered with Actinophytocola timorensisID05-A0653T (98.8 % 16S rRNA gene sequence similarity), Actinophytocola oryzaeGMKU 367T (98.6 %), Actinophytocola corallinaID06-A0464T (98.2 %) and Actinophytocola burenkhanensisMN08-A0203T (97.5 %). In addition, DNA-DNA hybridization values between BMP B8152T and A. timorensisID05-A0653T(44.2±3.6 %) and A. oryzaeGMKU 367T(36.7±2.3 %) were well below the 70 % limit for species identification. The combined phenotypic and genotypic data indicate that the isolate represents a novel species of the genus Actinophytocola, for which the name Actinophytocola glycyrrhizae sp. nov., is proposed, with the type strain BMP B8152T (=KCTC 49002T=CGMCC 4.7433T).

Actinomadura deserti sp. nov., isolated from desert soil

A Gram-positive, strictly aerobic actinobacterium, designated BMP B8004T, was isolated from desert soil collected in Xinjiang Province, Northwest China. It produced an extensively branched non-fragmenting substrate mycelium, and very scanty aerial mycelium that formed a short hooked chain of arthrospores with a smooth surface. Optimum growth occurred at 28 °C, pH 7.0 and 0 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain BMP B8004T formed a distinct phyletic lineage within the genus Actinomadura. It shared the highest 16S rRNA gene sequence similarity to Actinomadura apis IM17-1T (99.2 %) and Actinomadura rifamycini NBRC 14183T (98.6 %). However, it could be distinguished from the two closest strains based on the low levels of DNA-DNA relatedness (52.7±0.7 and 45±1.8 %, respectively). Chemotaxonomic characteristics, including the main phospholipids, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides, the major menaquinones MK-9(H6) and MK-9(H8), the predominant fatty acids iso-C16 : 0, C16 : 0, C18 : 0 10-methyl and C18 : 1ω9c, were also consistent with the properties of the genus Actinomadura. The DNA G+C content of strain BMP B8004T was 71.9 mol%. Based on phenotypic and genotypic features, strain BMP B8004T is considered to represent a novel species of the genus Actinomadura, for which the name Actinomadura deserti sp. nov. is proposed. The type strain is BMP B8004T (=CGMCC 4.7432T=KCTC 39998T).