Cell Wall Hydrolysates Research Articles

Streptomyces fuscus sp. nov., a brown-black pigment producing actinomycete isolated from dry mudflat sand.

A novel Gram-stain-positive, aerobic actinobacterial strain, designated GXMU-J15T, was isolated from dry mudflat sand. A polyphasic approach was employed for its taxonomic characterization. The strain developed extensively branched yellowish white to light yellow substrate mycelia and white aerial mycelia, and produced smooth cylindrical spores in a loose straight spore chain on International Streptomyces Project 2-7 agar media. Strain GXMU-J15T grew at 20-50 °C (optimum, 35 °C), at pH 5.0-8.0 (optimum, pH 7.0) and in the presence of 0-8 % (w/v) NaCl. Analysis of 16S rRNA gene sequences indicated that strain GXMU-J15T represents a member of the genus Streptomyces. Strain GXMU-J15T showed the highest 16S rRNA gene sequence similarity to Streptomyces lusitanus CGMCC 4.1745T (99.1 %) and Streptomyces thermocarboxydus CGMCC 4.1883T (98.8 %). Phylogenetic tree analysis based on multilocus sequence analysis (MLSA) and whole genome sequence construction revealed that strain GXMU-J15T was most closely related to Streptomyces cupreus PSKA01T, Streptomyces cinnabarinus DSM 40467T and Streptomyces davaonensis JCM 4913T. The MLSA and genome-to-genome distances between strain GXMU-J15T and its relatives were 0.0418, 0.0443 and 0.0485 and 0.1237, 0.1188 and 0.1179, respectively. The results of orthologous average nucleotide identity and digital DNA-DNA hybridization analysis corroborated the results of the MLSA and whole genome sequence evolution analysis, indicating that the novel isolate represents a distinct species of the genus Streptomyces. The whole-cell sugars of strain GXMU-J15T were xylose, glucose and galactose. The characteristic diamino acid in the cell-wall hydrolysate was ll-diaminopimelic acid. The lipids contained diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol, phosphatidylglycerides, phosphatidylcholine, two phospholipids of an unknown structure containing glucosamine, one unknown phospholipid and two unknown lipids. The major cellular fatty acid components were iso-C15 : 0, anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. The main respiratory quinone types were MK-9(H6) and MK-9(H8). The whole genome size of strain GXMU-J15T was 8.68 Mbp, with 71.23 mol% G+C content. Genomic analysis indicated that strain GXMU-J15T has the potential to synthesize polyketides, terpenes and a series of important antibiotics besides the gene cluster for melanin synthesis. Based on these genotypic and phenotypic data, strain GXMU-J15T is proposed to represent a new species of the genus Streptomyces named Streptomyces fuscus sp. nov. The type strain is GXMU-J15T (=MCCC 1K08211T=JCM 35917T).

Profundirhabdus halotolerans gen. nov., sp. nov., an haloalkaliphilic actinobacterium isolated from seawater of the Mariana Trench.

A Gram-stain-positive, strictly aerobic, rod-shaped actinobacterium, designated strain ZYF776T, was isolated from seawater of the Mariana Trench collected at a depth of 4000 m. Results of 16S rRNA gene sequence analysis indicated that strain ZYF776T was a member of the class Nitriliruptoria and closely related to Nitriliruptor alkaliphilus DSM 45188T (member of the order Nitriliruptorales, 94.94 % sequence similarity) and Egicoccus halophilus KCTC 33612T (member of the order Egicoccales, 94.46 %). Strain ZYF776T was catalase-positive and oxidase-negative. Growth occurred at 16-37 °C (optimum, 28 °C), in the presence of 0-13 % NaCl (w/v; optimum, 4 %) and at pH 7.0-10.0 (optimum, pH 8.0). Cell-wall hydrolysates of strain ZYF776T contained meso-diaminopimelic (peptidoglycan type A1γ), with ribose, rhamnose and a smaller amount of xylose as the cell-wall sugars. The major menaquinone was MK-10. The predominant fatty acids (>10 %) were C16:0, C17:1 ω8c and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c). The polar lipid profile mainly contained diphosphatidylglycerol, phosphatidylglycerol and phosphoglycolipid. The genomic DNA G+C content of strain ZYF776T was 68.7 mol%. The genome of strain ZYF776T was about 5.61 Mbp in size, which was larger than those of the reference strains N. alkaliphilus DSM45188T (5.56 Mbp) and E. halophilus KCTC 33612T (3.98 Mbp). The average nucleotide identity and digital DNA-DNA hybridization values between ZYF776T and the related strains N. alkaliphilus DSM 45188T and E. halophilus KCTC 33612T were 76.7 and 20.3 % and 75.8 and 20.0 %, respectively. Based on the polyphasic evidence, a novel genus and species with the name Profundirhabdus halotolerans gen. nov., sp. nov. is proposed. The type strain is ZYF776T (=JCM 33008T=MCCC 1K03555T).

Actinomadura spongiicola sp. nov., isolated from the marine sponge Leucetta chagosensis.

A novel actinomycete strain, named LHW52907T, was isolated from a marine sponge (Leucetta chagosensis) collected in the South China Sea. The strain developed branched mycelia without fragmentation and short spore chains in hook-and- spiral form with wrinkled surfaces, bearing no more 10 spores. The cell-wall hydrolysates contained meso-diaminopimelic acid as the diagnostic diamino acid. The sugars in whole-cell hydrolysates consisted of mannose, ribose, glucose, galactose and madurose. The major fatty acids of the strain were C16 : 0, C17 : 0 and C18 : 1 ω9c. The predominant menaquinone was MK-9(H6). The strain had the highest 16S rRNA gene sequence similarity of 99.72 % to Actinomadura pelletieri DSM 43383T. However, the average nucleotide identity and in silico DNA-DNA hybridization values between them were 93.6 and 52.6 %, respectively, readily distinguishing them as two different species. The results indicate that strain LHW52907T represents a novel species of the genus Actinomadura, for which we propose the name Actinomadura spongiicola sp. nov, with the type strain LHW52907T (=DSM 106571T=CGMCC 4.7596T).

Agromyces mariniharenae sp. nov., a novel indole-acetic acid producing actinobacterium isolated from marine sand.

A Gram-positive, aerobic, heterotrophic, non-endospore-forming, rod-shaped and indole-acetic acid-producing strain, designated NEAU-184T, was isolated from marine sand collected in Sanya, PR China, and its taxonomic position was investigated using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequence data indicated that strain NEAU-184T should be assigned to the genus Agromyces and formed a distinct branch with its closest neighbour, Agromyces iriomotensis NBRC 106452T (99.1 %). 2,4-Diaminobutyric acid, d-alanine, d-glutamic acid and glycine were detected in cell-wall hydrolysate and glucose, rhamnose and xylose were detected in whole-cell hydrolysate. The polar lipids were found to contain diphosphatidylglycerol, glycolipid, phosphatidylglycerol and two unidentified lipids. The major menaquinone was MK-12 and the minor menaquinones were MK-13 and MK-11. The predominant fatty acids were anteiso-C17 : 0, anteiso-C15 : 0 and iso-C16 : 0. The DNA G+C content was 71.5 mol%. Furthermore, the strain could be clearly distinguished from its closely related type strains by the combination of DNA-DNA hybridization results and some phenotypic characteristics. Meanwhile, the strain has the ability to produce indole-acetic acid (0.334mg ml-1). Therefore, strain NEAU-184T represents a novel species of the genus Agromyces, for which the name Agromyces mariniharenae sp. nov. is proposed, with strain NEAU-184T (=CGMCC 4.7505T=JCM 32546T) as the type strain.

Complete and efficient conversion of plant cell wall hemicellulose into high-value bioproducts by engineered yeast

Plant cell wall hydrolysates contain not only sugars but also substantial amounts of acetate, a fermentation inhibitor that hinders bioconversion of lignocellulose. Despite the toxic and non-consumable nature of acetate during glucose metabolism, we demonstrate that acetate can be rapidly co-consumed with xylose by engineered Saccharomyces cerevisiae. The co-consumption leads to a metabolic re-configuration that boosts the synthesis of acetyl-CoA derived bioproducts, including triacetic acid lactone (TAL) and vitamin A, in engineered strains. Notably, by co-feeding xylose and acetate, an enginered strain produces 23.91 g/L TAL with a productivity of 0.29 g/L/h in bioreactor fermentation. This strain also completely converts a hemicellulose hydrolysate of switchgrass into 3.55 g/L TAL. These findings establish a versatile strategy that not only transforms an inhibitor into a valuable substrate but also expands the capacity of acetyl-CoA supply in S. cerevisiae for efficient bioconversion of cellulosic biomass.

Rhodococcus spongiicola sp. nov. and Rhodococcus xishaensis sp. nov., from marine sponges

Two novel Rhodococcus strains, LHW50502T and LHW51113T, were isolated from marine sponges obtained on Xisha Island, Hainan Province, PR China. Rods and cocci, typical characteristics of the genus Rhodococcus, were observed. The strains contained meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall hydrolysates and galactose, arabinose, ribose and glucose as the whole-cell sugars. The major fatty acid identified was C16 : 0. MK-8(H4) was the predominat menaquinone of both strains. Stains LHW50502T and LHW51113T had almost identical (99.6 %) 16S rRNA gene sequences but shared relatively low similarities with previously characterized Rhodococcus species (well below 98.7 %). The results of phylogenetic analysis supported their closest relationship; however, the average nucleotide identity and digital DNA-DNA hybridization values between these two strains indicated that they belonged to distinct species. Taken together, the results of this study indicate that strains LHW50502T and LHW51113T represent two novel species of the genus Rhodococcus, for which the names Rhodococcus spongiicola sp. nov. (type strain LHW50502T=DSM 106291T=CCTCC AA 2018033T) and Rhodococcus xishaensis sp. nov. (type strain LHW51113T=DSM 106204T=CCTCC AA 2018034T) are proposed.

Enhanced 2′-Fucosyllactose production by engineered Saccharomyces cerevisiae using xylose as a co-substrate

2′-Fucosyllactose (2′-FL), a human milk oligosaccharide with confirmed benefits for infant health, is a promising infant formula ingredient. Although Escherichia coli, Saccharomyces cerevisiae, Corynebacterium glutamicum, and Bacillus subtilis have been engineered to produce 2′-FL, their titers and productivities need be improved for economic production. Glucose along with lactose have been used as substrates for producing 2′-FL, but accumulation of by-products due to overflow metabolism of glucose hampered efficient production of 2′-FL regardless of a host strain. To circumvent this problem, we used xylose, which is the second most abundant sugar in plant cell wall hydrolysates and is metabolized through oxidative metabolism, for the production of 2′-FL by engineered yeast. Specifically, we modified an engineered S. cerevisiae strain capable of assimilating xylose to produce 2′-FL from a mixture of xylose and lactose. First, a lactose transporter (Lac12) from Kluyveromyces lactis was introduced. Second, a heterologous 2′-FL biosynthetic pathway consisting of enzymes Gmd, WcaG, and WbgL from Escherichia coli was introduced. Third, we adjusted expression levels of the heterologous genes to maximize 2′-FL production. The resulting engineered yeast produced 25.5 g/L of 2′-FL with a volumetric productivity of 0.35 g/L∙h in a fed-batch fermentation with lactose and xylose feeding to mitigate the glucose repression. Interestingly, the major location of produced 2′-FL by the engineered yeast can be changed using different culture media. While 72% of the produced 2′-FL was secreted when a complex medium was used, 82% of the produced 2′-FL remained inside the cells when a minimal medium was used. As yeast extract is already used as food and animal feed ingredients, 2′-FL enriched yeast extract can be produced cost-effectively using the 2′-FL-accumulating yeast cells.

Nocardia macrotermitis sp. nov. and Nocardia aurantia sp. nov., isolated from the gut of the fungus-growing termite Macrotermes natalensis.

The taxonomic positions of two novel aerobic, Gram-stain-positive Actinobacteria, designated RB20T and RB56T, were determined using a polyphasic approach. Both were isolated from the fungus-farming termite Macrotermes natalensis. Results of 16S rRNA gene sequence analysis revealed that both strains are members of the genus Nocardia with the closest phylogenetic neighbours Nocardia miyunensis JCM12860T (98.9 %) and Nocardia nova DSM44481T (98.5 %) for RB20T and Nocardia takedensis DSM 44801T (98.3 %), Nocardia pseudobrasiliensis DSM 44290T (98.3 %) and Nocardia rayongensis JCM 19832T (98.2 %) for RB56T. Digital DNA–DNA hybridization (DDH) between RB20T and N. miyunensis JCM12860T and N. nova DSM 44481T resulted in similarity values of 33.9 and 22.0 %, respectively. DDH between RB56T and N. takedensis DSM44801T and N. pseudobrasiliensis DSM44290T showed similarity values of 20.7 and 22.3 %, respectively. In addition, wet-lab DDH between RB56T and N. rayongensis JCM19832T resulted in 10.2 % (14.5 %) similarity. Both strains showed morphological and chemotaxonomic features typical for the genus Nocardia , such as the presence of meso-diaminopimelic acid (A2pm) within the cell wall, arabinose and galactose as major sugar components within whole cell-wall hydrolysates, the presence of mycolic acids and major phospholipids (diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol), and the predominant menaquinone MK-8 (H4, ω-cyclo). The main fatty acids for both strains were hexadecanoic acid (C16 : 0), 10-methyloctadecanoic acid (10-methyl C18 : 0) and cis-9-octadecenoic acid (C18 : 1 ω9c). We propose two novel species within the genus Nocardia : Nocardia macrotermitis sp. nov. with the type strain RB20T (=VKM Ac-2841T=NRRL B65541T) and Nocardia aurantia sp. nov. with the type strain RB56T (=VKM Ac-2842T=NRRL B65542T).

Planococcus lenghuensis sp. nov., an oil-degrading bacterium isolated from petroleum-contaminated soil.

A Gram-staining-positive and aerobic coccus with the ability to degrade petroleum bacterium, designated Y42T, was isolated from the Lenghu oil field located in the northern margin of the Qaidam Basin. Phylogenetic and signature nucleotides analyses revealed that strain Y42T belongs to the genus Planococcus. The multiple sequence alignments of 16S rRNA and housekeeping genes showed that strain Y42T formed a distinct lineage with the other Planococcus clade. The average nucleotide identity (ANI) and DNA-DNA hybridization values (DDH) between strain Y42T and the reference strains were 69.5-70.1 and 19.4-21.7%, respectively, which values were below the threshold for species delineation. The major fatty acids of strain Y42T were anteiso-C15:0. The respiratory quinone was MK-7 (71.8%) as the predominant menaquinone followed the MK-6 (28.2%) and the cell-wall hydrolysates contained LL-diaminopimelic acid. The polar lipid was composed of diphosphatidyl glycerol, phosphatidyl glycerol, phosphoglycolipid, aminophospholipid and four unidentified lipids. The peptidoglycan type was A4α (L-Lys-D-Glu). The strain Y42T possessed larger genome (approximately 4MB) and revealed obvious differences for the abundance of the COG categories compared with the other Planococcus bacteria. Also, the strain Y42T also possessed more unique orthologous proteins. The structural characteristics of the strain Y42T genome provided a competitive advantage for better survival in petroleum-polluted environments. Combined with the 16S rRNA gene and genome sequence, phenotypic as well as chemotaxonomic characterisations, strain Y42T is considered to represent a novel species of the genus Planococcus, for which the name Planococcus lenghuensis sp. nov. be proposed. The type strain is Y42T (= CGMCC 1.15921T = JCM 32719T).

Saccharothrix deserti sp. nov., an actinomycete isolated from desert soil.

A Gram-stain-positive, aerobic actinomycete, designated strain BMP B8144T, was isolated from desert soil, in Xinjiang province, northwest China. The isolate produced scanty aerial mycelium and fragmented substrate mycelium on most tested media. Cell-wall hydrolysates contained meso-diaminopimelic acid, galactose and mannose. The diagnostic phospholipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylhydroxylethanolamine, phosphatidylinositol, and phosphatidylinositol mannosides. The major fatty acids included iso-C16 : 0, C17 : 1 ω8c and iso-C15 : 0. The predominant menaquinones were MK-9(H4) and MK-10(H4). The DNA G+C content was 70.4 mol% (genome). Based on the 16S rRNA gene sequence analysis on EzBioCloud server, strain BMP B8144T showed the closest similarities to Saccharothrix lopnurensis YIM LPA2hT (98.9 %) and 'Saccharothrix yanglingensis' Hhs.015 (98.6 %). However, it can be distinguished from the closest strains based on the low levels of DNA-DNA relatedness (59.3±1.8 and 47.9±2.3 %, respectively). A combination of morphological, chemotaxonomic and phylogenetic characteristics, strain BMP B8144T represents a novel species of the genus Saccharothrix, for which the name Saccharothrix deserti sp. nov. is proposed. The type strain is BMP B8144T (=CGMCC 4.7490T=KCTC 49001T).

Synthesis of a Di-Mycoloyl Tri-Arabinofuranosyl Glycerol Fragment of the Mycobacterial Cell Wall, Based on Synthetic Mycolic Acids.

Fragments of mycobacterial cell walls such as arabinoglycerol mycolate and dimycoloyl diarabinoglycerol, comprising complex mixtures of mycolic acids, have immunostimulatory and antigenic properties. A related di-mycoloyl tri-arabinofuranosyl glycerol fragment has been isolated from cell wall hydrolysates. An effective stereoselective synthesis of tri-arabinofuranosyl glycerol, followed by coupling with stereochemically defined mycolic acids of different structural classes, to provide unique di-mycoloyl tri-arabinofuranosyl glycerols is now described.

Gandjariella thermophila gen. nov., sp. nov., a new member of the family Pseudonocardiaceae, isolated from forest soil in a geothermal area

Six mycelium-forming actinomycete strains were isolated from forest soil near the Cisolok geysers in West Java, Indonesia. The 16S rRNA gene sequences of these strains showed high similarity to members of genera in the family Pseudonocardiaceae with values less than 96.0 %, and most closely related to the genus Thermotunica, T. guangxiensis AG2-7T(94.6-95.2 % similarity). The type strain, designated SL3-2-4T, was aerobic, thermophilic, Gram-stain-positive that formed branched, non-fragmented substrate mycelia and unbranched aerial mycelia with long-chain, oval-shaped spores on International Streptomyces Project (ISP) 3 medium. It produced light-orange substrate mycelia and light-orange diffusible pigments on ISP 3 medium with 2 % gellan gum, grown at 30-55 °C, with optimum growth at 45 °C. The pH range for growth was 4.0-8.0, with optimum growth at pH 7.0. Strain SL3-2-4T was able to hydrolyze casein, esculin, gelatin, guanine, hypoxanthine, starch, L-tyrosine, and xanthine, but not adenine, carboxymethyl-cellulose, cellulose, chitin, Tween 20, or xylan. The major fatty acid was iso-C16 : 0, and the major menaquinone was MK-8 (H4). The detected polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidyl-N-methylethanolamine, unidentified aminophospholipids, unidentified glycolipids, and unidentified phospholipids. The cell wall hydrolysate of SL3-2-4T contained meso-2,4-diaminopimelic acid. The whole cell sugars were arabinose and galactose. The DNA G+C content was 71.6 mol%. Phenotypic features and phylogenetic data differentiated SL3-2-4T from members of the family Pseudonocardiaceae. Therefore, the strain SL3-2-4T is proposed as a representative of a novel species in a novel genus, Gandjariella thermophila gen. nov., sp. nov. The type strain is SL3-2-4T (=UICC B-83T=NRRL B-67478T=InaCC A981T).

Streptomyces cyaneochromogenes sp. nov., a blue pigment-producing actinomycete from manganese-contaminated soil.

A novel actinomycete isolate, designated strain MK-45T, was isolated from soil sampled at a manganese-contaminated area in Xiangtan, China. The isolate formed extensively branched substrate mycelia and aerial mycelia that differentiated into tightly coiled or spiral spore chains with smooth-surfaced spores. The cell-wall hydrolysates contained ll-diaminopimelic acid and traces of meso-diaminopimelic acid. The major menaquinones consisted of MK-9(H4), MK-9(H6) and MK-9(H8). The major polar lipids contained diphosphatidylgycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphotidylinositol and phosphotidylinositol mannosides. The predominant cellular fatty acids were iso-C16 : 0, C16 : 0 and summed feature 3. Phylogenetic analysis based on the 16S rRNA gene sequences and concatenated partial sequences of the five protein-coding genes indicated that strain MK-45T was a member of the genus Streptomyces and most closely related to Streptomyces caeruleatus GIMN4.002T (99.5 % similarity), Streptomyces curacoi CGMCC 4.1680T (99.5 %), Streptomyces shaanxiensis CCNWHQ 0031T (99.4 %) and Streptomyces cinnabarinus NRRL B12382T (98.9 %), respectively. However, the digital DNA-DNA hybridization values, the average nucleotide identity values and MLSA evolutionary distances between strain MK-45T and them showed that it belonged to a distinct species. Furthermore, the results of morphological, physiological and biochemical tests allowed further phenotypic differentiation of strain MK-45T from the four closely related type strains mentioned above and other species of the genus Streptomyces with which this strain has 98.0-99.2 % 16S rRNA gene sequence similarity. Therefore, it is concluded that strain MK-45T represents a novel species of the genus Streptomyces, for which the name Streptomycescyaneochromogenes sp. nov. is proposed, with MK-45T (=CICC 11045T=KCTC 49099T) as the type strain.

Streptomyces fodineus sp. nov., an actinobacterium with antifungal activity isolated from mine area soil.

A novel actinobacterial strain producing an antifungal substance was isolated from a sample of acidic mine area soil, and its taxonomic position was evaluated. The novel strain, designated TW1S1T, formed white-grey aerial mycelium and yellow substrate mycelium on oatmeal agar. Growth occurred at 10-45 °C (optimum, 30 °C), pH 4-9 (pH 6-7) and in the presence of up to 8 % (w/v) NaCl. Melanin was produced on peptone-yeast extract-iron agar. Phylogenetic analysis based on its 16S rRNA gene sequence indicated that the novel strain should be assigned to the genus Streptomyces, and the closest species was Streptomyces puniciscabiei S77T with 99.1 % sequence similarity, which was followed by Streptomyces durhamensis NRRL B-3309T (99.0 %), Streptomyces filipinensis NBRC 12860T (98.9 %) and Streptomyces yaanensis Z4T (98.7 %). The chemotaxonomic properties were consistent with those of Streptomyces. ll-Diaminopimelic acid was the diagnostic diamino acid, and alanine, glutamic acid and glycine were present in the peptidoglycan. The cell-wall hydrolysate also contained galactose, glucose, mannose and ribose. The predominant isoprenoid quinones were MK-9(H4) and MK-9(H6), the major polar lipids were phosphatidylglycerol and an unidentified phospholipid, and the main fatty acids were iso-C16 : 0 and anteiso-C15 : 0. However, strain TW1S1T could be distinguished from its neighbouring species by its phenotypic properties. In addition, the genome-based comparison with the closest species indicated that strain TW1S1T should be recognized as a separate species. The phylogenetic, phenotypic and chemotaxonomic as well as genomic evidence supported that TW1S1T represents a novel species of Streptomyces, for which the name Streptomycesfodineus sp. nov. is proposed (type strain, TW1S1T = KCTC 49013T = JCM 32404T).

Actinomadura craniellae sp. nov., isolated from a marine sponge in the South China Sea.

A novel marine actinomycete, designated LHW63021T, was isolated from a marine sponge, genus Craniella, collected in the South China Sea. A polyphasic approach was applied to characterize the taxonomic position of this strain. The strain was found to have scarce aerial mycelia that differentiated into spore chains. The cell-wall hydrolysates contained meso-diaminopimelic acid as the diagnostic diamino acid. Glucose, galactose, mannose and madurose were found in the whole-cell hydrolysates. The dominant polar lipids were phosphatidylinositol and diphosphatidylglycerol. MK-9(H6) and MK-9(H8) were the predominant menaquinones. The major fatty acids were iso-C16 : 0, iso-C18 : 0, 10-methyl C17 : 0 and C18 : 1 ω9c. The DNA G+C content based on the draft genome sequence was 72.0 mol%. 16S rRNA gene sequence analysis indicated that strain LHW63021T was a member of the genus Actinomadura and had the highest similarity to Actinomadura echinospora DSM 43163T (97.3 %). Phylogenetic trees supported their close relationship. The average nucleotide identity and digital DNA-DNA hybridization values between the whole genome sequences of strain LHW63021T and A. echinospora DSM 43163T were 79.13 and 23.20 %, respectively. The evidence from the polyphasic study shows that strain LHW63021T represents a novel species of the genus Actinomadura, for which the name Actinomadura craniellae sp. nov. is proposed. The type strain is LHW63021T (=DSM 106125T=CCTCC AA 2018015T).

Streptomyces cadmiisoli sp. nov., a novel actinomycete isolated from cadmium-contaminated soil.

A novel Streptomyces strain, ZFG47T, isolated from a cadmium-contaminated soil sample, was taxonomically studied in detail. Strain ZFG47T formed long, flexuous spiral spore chains consisting of elliptoid spores with spiny surfaces. The cell-wall hydrolysates contained ll-diaminopimelic acid as the diagnostic diamino acid. The major menaquinones consisted of MK-9(H2), MK-9(H4) and MK-9(H8). The major polar lipids contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylinositol mannosides. The predominant cellular fatty acids were iso-C16 : 0, C16 : 0 and anteiso-C15 : 0. The 16S rRNA gene sequence-based phylogenetic analysis indicated that this strain belongs to the genus Streptomyces, showing the highest sequence similarity to Streptomyceskoyangensis VK-A60T (98.7 %). However, the digital DNA-DNA hybridization value, the average nucleotide identity value and the MLSA evolutionary distance between this strain and S. koyangensis VK-A60T showed that it belonged to a distinct species. Furthermore, the novel isolate could be distinctly differentiated from S. koyangensis VK-A60T by morphological, physiological and biochemical characteristics. On the basis of the evidence from this polyphasic study, it is concluded that strain ZFG47T represents a novel species of the genus Streptomyces, for which the name Streptomyces cadmiisoli sp. nov. is proposed, with strain ZFG47T (CICC 11050T=JCM 32897T) as the type strain.

Nocardia mangyaensis sp. nov., a novel actinomycete isolated from crude-oil-contaminated soil.

A Gram-stain-positive, aerobic, non-motile and mycolic-acid-containing strain, designated Y48T, was isolated from soil contaminated by crude oil located in the northern margin of the Qaidam Basin. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Y48T belongs to the genus Nocardia and is closely related to N. cummidelens DSM 44490T (99.0 % similarity), N. soli DSM 44488T (99.0 %), N. lasii 3C-HV12T (98.9 %), N. salmonicida NBRC 13393T (98.6 %), N. ignorata NBRC 108230T (98.6 %) and N. coubleae NBRC 108252T (98.6 %). The average nucleotide identity and DNA-DNA hybridization values between strain Y48T and the reference strains were 75.9-84.5 and 27.5-29.0 %, respectively, values that were below the thresholds for species delineation. Chemotaxonomic analysis indicated that the major fatty acids of strain Y48T were C16 : 0, summed feature 3 (C16 : 1ω6c/C16 : 1ω7c), C18 : 1ω9c and C18 : 0 10-methyl (TBSA). The respiratory quinone was MK-8(H4, ω-cycl). The polar lipid profile was composed of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside, two glycolipids and three unidentified lipids. The cell-wall hydrolysates contained meso-diaminopimelic acid, with ribose, arabinose, glucose and galactose as whole-cell sugars. A combination of 16S rRNA gene sequence analysis, and phenotypic and chemotaxonomic characterizations demonstrated that strain Y48T represents a novel species of the genus Nocardia, for which the name Nocardia mangyaensis sp. nov. is proposed. The type strain is Y48T (=JCM 32795T=CGMCC 4.7494T).

Streptomyces reniochalinae sp. nov. and Streptomyces diacarni sp. nov., from marine sponges.

Two marine actinomycete strains, LHW50302T and LHW51701T, were isolated from marine sponges collected in Sansha, Hainan Province, China. The morphological, chemotaxonomic and phylogenetic characteristics were consistent with their classification in the genus Streptomyces. The strains formed hooked and looped chains of arthrospores with smooth surfaces. The cell-wall hydrolysates of the strains contained ll-diaminopimelic acid as the diagnostic diamino acid. MK-9(H8) was the predominant menaquinone. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. Major fatty acids of the strains were iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The 16S rRNA gene sequences indicated that the strains clustered together with Streptomyces albus CGMCC 4.1640T and Streptomyces qinglanensis CGMCC 4.6825T. Multilocus sequence analysis (MLSA) confirmed their relationship. Genome relatedness in forms of average nucleotide identity, digital DNA-DNA hybridization value and MLSA evolutionary distance between each of the strains and its closest relatives showed that they belonged to distinct species. On the basis of these results, strains LHW50302T and LHW51701T belong to two novel species in the genus Streptomyces, for which the names Streptomyces reniochalinae sp. nov. (type strain LHW50302T=CCTCC AA 2018013T=DSM 106194T) and Streptomyces diacarni sp. nov. (type strain LHW51701T=CCTCC AA 2018017T=DSM 106126T) are proposed, respectively.

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).

Naumannella huperziae sp. nov., an endophytic actinobacterium isolated from Huperzia serrata (Thunb.)

An endophytic, short rod-shaped, non-motile and non-spore-forming actinobacterium, designated strain CPCC 204135T, was isolated from a surface-sterilized medicinal plant, Huperzia serrata (Thunb.), collected from Sichuan Province, south-west China. Strain CPCC 204135T was observed to grow at temperatures between 20 and 37 °C (optimum, 28-32 °C), at pH 6.0-9.0 (optimum, pH 7.0-8.0) and in the presence of 0-9.0 % (w/v) NaCl (optimum, 0-3 %). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CPCC 204135T belonged to the genus Naumannella, showing the highest level of 16S rRNA gene sequence similarity with Naumannella halotolerans DSM 24323T (97.2 %), the only species of the genus Naumannella in the family Propionibacteriaceaewith avalidly published name. The DNA-DNA hybridization value between strain CPCC 204135T and N. halotolerans DSM 24323T was 20.1±1.8 %, which is far below the accepted 70 % threshold for species delineation. The cell-wall hydrolysates contained ll-diaminopimelic acid, alanine, glycine and glutamic acid, with the peptidoglycan type of A3γ. Diphosphatidylglycerol, phosphatidylglycerol, an unidentified phospholipid, one unidentified aminolipid, one unidentified polar lipid and several kinds of glycolipids were detected in the polar lipids profile. MK-9(H4) was identified as the predominant menaquinone. The major cellular fatty acids (>10 %) were anteiso-C15 : 0 and iso-C16 : 0. The G+C content of the genomic DNA of strain CPCC 204135T was determined to be 71.8 mol%. On the basis of phylogenetic analysis, and phenotypic and chemotaxonomic characteristics, we concluded that strain CPCC 204135T represents a novel species of the genus Naumannella, for which the name Naumannella huperziae sp. nov. is proposed, with strain CPCC 204135T (=DSM 101717T=NBRC 111773T) as the type strain.