Chromosome topology and genome size of selected actinomycetes species

Information about the genome organization of actinomycetes species is restricted to a few genera: Corynebacterium, Mycobacterium, Rhodococcus, Saccharopolyspora and Streptomyces. Streptomyces species and Saccharopolyspora erythraea were shown to contain a single linear 8 Mb chromosome. In contrast, the Corynebacterium, Mycobacterium and Rhodococcus species studied were demonstrated to possess a smaller (3 Mb–6.5 Mb) single circular chromosome. To investigate whether linear chromosome topology and genome sizes above 7 Mb are unique features of Streptomyces and S. erythraea we have started to investigate the chromosome topology, the genome size and the status of accessory elements of additional actinomycetes species: Actinoplanes philippinensis, Amycolatopsis orientalis, Micromonospora chalcea, Nocardia asteroides, Rhodococcus opacus and Streptoverticillium abikoense. Our data which are based on PFGE experiments clearly suggest that large genome sizes and chromosome linearity are seen in mycelium forming actinomycetes genera. In addition we have identified large linear plasmids in Nocardia asteroides, Streptoverticillium abikoense and Rhodococcus opacus.     

In vivo evolution of the Rhodococcus sp. strain DS7: selection of recombinants able to desulfurize both dibenzothiophene and benzothiophene

Rhodococcus sp. DS7, isolated from a polluted soil, has shown good desulfurizing activity towards dibenzothiophene (DBT) and its derivatives, but is not able to desulfurize benzothiophene (BT), the other thiophenic molecule recalcitrant to the chemical hydrodesulfurization (HDS) process, and most abundant in gasoline. To select a Rhodococcus DS7 derivative strain able to desulfurize both DBT and BT, we took advantage of the verified capacity of this strain to integrate exogenous DNA randomly, with a good efficiency. Heterologous chromosomal DNA, digested with restriction enzymes, from two BT but not DBT desulfurizing strains, Rhodococcus sp. ATCC 27778 and Gordonia sp. ATCC 19067, was electroporated into Rhodococcus DS7. Selection on minimal medium with BT as sole sulfur source allowed us to isolate several DS7 derivatives with the capacity to desulfurize both thiophenic molecules. Two strains, one derived from the integration and recombination of DNA from ATCC 27778, and the other from ATCC 19067, have been partially characterized. These recombinant microorganisms are an interesting starting point to develop new biodesulfurization processes.     

Microbial degradation of 7-ketocholesterol

7-Ketocholesterol (7KC) is an oxidized derivative of cholesterol suspected to be involved in the pathogenesis of atherosclerosis and possibly Alzheimer’s disease. While some oxysterols are important biological mediators, 7KC is generally cytotoxic and interferes with cellular homeostasis. Despite recent interest in preventing the accumulation of 7KC in a variety of matrices to avoid adverse biological effects, its microbial degradation has not been previously addressed in the peer-reviewed literature. Thus, the rate and extent of biodegradation of this oxysterol was investigated to bridge this gap. A wide variety of bacteria isolated from soil or activated sludge, including Proteobacterium Y-134, Sphingomonas sp. JEM-1, Nocardia nova, Rhodococcus sp. RHA1, and Pseduomonas aeruginosa, utilized 7KC as a sole carbon and energy source, resulting in its mineralization. Nocardia nova, which is known to produce biosurfactants, was the fastest degrader. This study supports the notion that microbial catabolic enzymes could be exploited to control 7KC levels in potential biotechnological applications for agricultural, environmental, or medical use.     

ESR Spectral Studies on the Free Radical Formed by the Reaction of Dehydroascorbic Acid with Amino Acid

Acrylonitrile-hydrating activity was found in various bacteria belonging to the genera Arthrobacter, Corynebacterium, Microbacterium, Nocardia, Pseudomonas and Rhodococcus. A strain, N-774, isolated by acetonitrile enrichment culture from soil showed the highest activity. Taxonomic studies indicated that strain N-774 belonged to the genus Rhodococcus. The acrylonitrile-hydrating enzyme of strain N-774 was constitutively formed in the cells. Besides acrylonitrile, many nitriles were hydrated to the corresponding amides. «-Butyronitrile, suc- cinonitrile and chloroacetonitrille served as suitable substrates. This bacterium could utilize many aliphatic nitriles and amides as a sole source of nitrogen but hardly utilized malononitrile, acrylonitrile or acrylamide. Cells having high nitrile hydratase activity (about 50units/mg of dry cells) could be easily prepared by cultivation at 30°C for 40 hr in a malt extract and yeast extract medium.     

Studio di una nuova specie di Streptomyces: Streptomyces nobilis sp. nov. e esame di ceppi affini appartenenti ai generi streptomyces,Streptoverticillium e Nocardia

Riassunto Alcuni ceppi di attinomiceti provenienti da diverso isolamento sono stati esaminati in comparazione con altri riferiti a specie o generi noti, onde procedere alla loro identificazione. I criteri diagnostici sono gli stessi di cui a precedenti nostre pubblicazione.I ceppi in studio sono riferibili a tre generi come da seguito: GenereStreptomyces. Sono riferiti aStreptomyces i ceppi 631 e 725, isolati da suolo, e sono descritti come specie nuovaStreptomyces nobilis sp. nov. Il ceppo 631 costituisce l'olotipo e il ceppo 725 una variante naturale.GenereStreptoverticillium. Viene riferito il ceppo in studio 174. Si è rinunziato ad una identificazione specifica limitando si all'inserimento nella serie Rubrireticuli della quale viene aggiornata la chiave delle specie.GenereNocardia. I ceppi 608 e 669 sono riferiti aNocardia asteroides e il ceppo 535 aProactinomyces (=Nocardia)ruber.

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Summary Strains of actinomycetes, isolated from different sources, have been examined comparatively together with cultures of known species and genera, in order to proceed to their identification. The diagnostic criteria followed are those described in previous papers.Our isolates have been referred to the following genera:GenusStreptomyces: to this genus belong our strains 631 and 725, isolated from soil and described as new species,Streptomyces nobilis sp. nov. Strain 631 is considered as the holotype and strain 725 as a natural variant.GenusStreptoverticillium: strain 174 belongs to this genus. No specific identification has been given, but the strain has been enclosed in the Rubrireticuli Series. A new key of the Series is provided.GenusNocardia: strains 608 and 669 are referred toNocardia asteroides, strain 535 toProactinomyces (=Nocardia)ruber.

     

Defluviimonas denitrificans gen. nov., sp. nov., and Pararhodobacter aggregans gen. nov., sp. nov., non-phototrophic Rhodobacteraceae from the biofilter of a marine aquaculture

Three Gram-negative bacterial strains were isolated from the biofilter of a recirculating marine aquaculture. They were non-pigmented rods, mesophiles, moderately halophilic, and showed chemo-organoheterotrophic growth on various sugars, fatty acids, and amino acids, with oxygen as electron acceptor; strains D9-3^T and D11-58 were in addition able to denitrify. Phototrophic or fermentative growth could not be demonstrated. Phylogenetic analysis of the 16S rRNA gene sequences placed D9-3^T and D11-58, and D1-19^T on two distinct branches within the alpha-3 proteobacterial Rhodobacteraceae, affiliated with, but clearly separate from, the genera Rhodobacter, Rhodovulum, and Rhodobaca. Based on morphological, physiological, and 16S rRNA-based phylogenetic characteristics, the isolated strains are proposed as new species of two novel genera, Defluviimonas denitrificans gen. nov., sp. nov. (type strain D9-3^T = DSM 18921^T = ATCC BAA-1447^T; additional strain D11-58 = DSM19039 = ATCC BAA-1448) and Pararhodobacter aggregans gen. nov., sp. nov (type strain D1-19^T = DSM 18938^T = ATCC BAA-1446^T).     

Characterization of selected actinomycetes degrading polyaromatic hydrocarbons in liquid culture and spiked soil

Summary Five strains of the Rhodococcus and Gordonia genera were evaluated for their potential use in bioremediation of polycyclic aromatic hydrocarbons (PAH) with or without another substrate (co-substrate). Their ability to produce biosurfactants or to degrade phenanthrene when growing on glucose, hexadecane and rapeseed oil was tested in liquid medium at 30 °C. All strains showed biosurfactant activity. The highest reduction in surface tension was recorded in whole cultures of Rhodococcus sp. DSM 44126 (23.1%) and R. erythropolis DSM 1069 (21.1%) grown on hexadecane and Gordonia sp. APB (20.4%) and R. erythropolis TA57 (18.2%) grown on rapeseed oil. Cultures of Gordonia sp. APB and G. rubripertincta formed emulsions when grown on rapeseed oil. After 14 days of incubation, Rhodococcus sp. DSM 44126 degraded phenanthrene (initial concentration 100 μg ml⁻¹) as sole carbon source (79.4%) and in the presence of hexadecane (80.6%), rapeseed oil (96.8%) and glucose (below the limit of detection). The other strains degraded less than 20%, and then with a co-substrate only. Rhodococcus sp. DSM 44126 was selected and its performance evaluated in soil spiked with a mixture of PAH (200 mg kg⁻¹). The effect of the addition of 0, 0.1 and 1% rapeseed oil as co-substrate was also tested. Inoculation enhanced the degradation of phenanthrene (55.7% and 95.2% with 0.1% oil and without oil respectively) and of anthracene (29.2% with 0.1% oil). Approximately 96% of anthracene and 62% of benzo(a)pyrene disappeared from the soil (inoculated and control) after 14 days and anthraquinone was detected as a metabolite. Rhodococcus sp. DSM 44126 was identified as Rhodococcus wratislaviensis by 16S rRNA sequencing and was able to degrade anthracene as sole carbon source in liquid culture.     

Characterization of clinical isolates of pathogenic <Emphasis Type="Italic">Nocardia</Emphasis> strains and related actinomycetes in Thailand from 1996 to 2003

In Thailand from 1996 to 2003, 171 strains of pathogenic aerobic actinomycetes from clinical specimens were isolated. Of those strains, 134 were mycolic acid containing actinomycetes, including 96 strains of Nocardia species. Others included 10 strains of Gordonia, 14 strains of Rhodococcus, and 22 strains of Mycobacterium. One strain each of the genera Tsukamurella and Corynebacterium were also isolated. Also identified were 27 strains of non-mycolic acid containing actinomycetes. Our identification studies of 96 strains of Nocardia species showed that significant pathogens in Thailand were N. beijingensis (18 strains), N. cyriacigeorgica (13 strains), and N. farcinica (34 strains); the most prevalent species was N. farcinica (35.4%). We also isolated four strains of N. asiatica, five strains of N. asteroides sensu stricto, four strains of N. nova, seven strains of N. otitidiscaviarum, eight strains of N. transvalensis, and two strains of N. pseudobrasiliensis.     

Prescottia equi gen. nov., comb. nov.: a new home for an old pathogen

The taxonomic status of Rhodococcus equi, originally isolated from foal specimens, has been the subject of discussion for a number of years. The chequered history of the taxon has prompted this polyphasic analysis of R. equi strains, close members of the genus Rhodococcus and representatives of other genera classified in the order Corynebacteriales, to establish the taxonomic position of this taxon. Thirty one R. equi strains, including the type strain, were examined for genotypic and numerical taxonomic properties. The resultant data are consistent with their classification in the order Corynebacteriales but the R. equi strains formed a distinct phyletic clade away from representatives of other members of the genus Rhodococcus in the 16S rRNA gene tree. Representatives of this clade shared their highest pairwise 16S rRNA gene sequence similarities with the type strain of Rhodococcus kunmingensis (95.2–98.1 %). However, the R. equi taxon was readily distinguished from R. kunmingensis and from the other members of the order Corynebacteriales using a combination of genotypic, chemotypic and phenotypic properties. On the basis of these data the R. equi strains are considered to represent a new genus. The name proposed for this taxon is Prescottia gen. nov., with Prescottia equi comb. nov. as the type species containing the type strain, C 7^T (= ATCC 25729^T = ATCC 6939^T = CCUG 892^T = CIP 54.72^T = DSM 20307^T = HAMBI 2061^T = NBRC 14956^T = JCM 1311^T = JCM 3209^T = LMG 18452^T = NBRC 101255^T = NCTC 1621^T = NRRL B-16538^T = VKM Ac-953^T).     

Three new anascosporic genera of the Saccharomycotina: Danielozyma gen. nov., Deakozyma gen. nov. and Middelhovenomyces gen. nov

Three new non-ascosporic, ascomycetous yeast genera are proposed based on their isolation from currently described species and genera. Phylogenetic placement of the genera was determined from analysis of nuclear gene sequences for D1/D2 large subunit rRNA, small subunit rRNA, translation elongation factor-1α and RNA polymerase II, subunits B1 and B2. The new taxa are: Deakozyma gen. nov., type species Deakozyma indianensis sp. nov. (type strain NRRL YB-1937, CBS 12903); Danielozyma gen. nov., type species Danielozyma ontarioensis comb. nov. (type strain NRRL YB-1246, CBS 8502); D. litseae comb. nov. (type strain NRRL YB-3246, CBS 8799); Middelhovenomyces gen. nov., type species Middelhovenomyces tepae comb. nov. (type strain NRRL Y-17670, CBS 5115) and M. petrohuensis comb. nov. (type strain NRRL Y-17663, CBS 8173).     

Numerical classification of some Rhodococci, Corynebacteria and related organisms

Nineteen strains of Corynebacterium sensu stricto, 23 received as Corynebacterium equi or Rhodococcus equi, marker cultures of Arthrobacter, Brevibacterium, Bacterionema matruchotii, Cellulomonas flavigena, Kurthia zopfii, Listeria denitrificans, Microbacterium lacticum, Rhodococcus rubropertinctus and 88 representatives of Mycobacterium, Nocardia, Rhodococcus and the 'aurantiaca' taxon were the subject of numerical phenetic analyses using 92 characters. The data were examined using the simple matching (SSM) and Jaccard (SJ) coefficients and clustering was achieved using the average linkage algorithm. With a single exception, strains containing meso-diaminopimelic acid, arabinose, galactose and mycolic acids were recovered in five aggregate clusters corresponding to Corynebacterium sensu stricto, Mycobacterium, Nocardia, Rhodococcus and the 'aurantiaca' taxon. Most of the Corynebacterium (Rhodococcus) equi strains formed a good taxospecies which included the type strain of Corynebacterium hoagii. The numerical data, and the results of earlier chemical and genetical studies, also provide sufficient evidence for the transfer of Bacterionema matruchotii to Corynebacterium sensu stricto as Corynebacterium matruchotii comb.nov. and for the recognition of Rhodococcus globerulus sp.nov. for some strains previously classified as Rhodococcus rubropertinctus (Hefferan) Goodfellow & Alderson. The classification of the remaining marker strains correlates well with other major developments in coryneform taxonomy.     

Enantioselective Hydrolysis of Butyl 2-Ethylhexanoate by a Strain of <Emphasis Type="Italic">Nocardia corynebacteroides</Emphasis>

The results of a screen for microbial esterases that have enantioselective activity for the hydrolysis of butyl 2-ethylhexanoate are described. The preliminary screen determined that a nocardioform bacterial strain, NRRL 21057, exhibited significant activity in preferentially hydrolyzing the S enantiomer of butyl 2-ethylhexanoate. Molecular systematics methods identified NRRL 21057 as a strain of Nocardia corynebacteroides. A survey of phylogenetically related species in the genera Gordonia, Rhodococcus, and Nocardia strains demonstrated that N. corynebacteroides NRRL 21057 is the most active strain known for the specific hydrolysis of the R-isomer of butyl 2-ethylhexanoate and that it provides the S-isomer of 2-ethylhexanoate in 86% enantiomeric excess within 22 h.     

Cloning vector system forNocardia spp.

An electro-transformation system has been developed forNocardia asteroides andNocardia corallina by using aNocardia-Escherichia coli shuttle vector. The shuttle vector, named pCY104, was constructed by joining a 2.5-kb crypticN. asteroides plasmid pCY101 with theE. coli plasmid pIJ4625. The resistance genes for kanamycin, chloramphenicol, and thiostrepton on plasmid pCY104 were expressed inN. asteroides andN. corallina. The transformation method was optimized forN. asteroides, and transformation efficiency of 8×10⁴ transformants per g plasmid DNA was achieved routinely.     

Antrihabitans stalagmiti sp. nov., isolated from a larva cave and a proposal to transfer Rhodococcus cavernicola Lee et al. 2020 to a new genus Spelaeibacter as Spelaeibacter cavernicola gen. nov. comb. nov

An actinobacterial strain, designated YC3-6^T, was isolated from a larva cave in Jeju, Republic of Korea. The novel isolate was found to grow at 10–30 °C, pH 5.0–10.0 and 0–4% (w/v) NaCl. The 16S rRNA gene phylogeny showed that the novel isolate formed a distinct subline within the family Nocardiaceae. Levels of 16S rRNA gene similarity indicated that the close relatives are Rhodococcus cavernicola (98.4% sequence similarity) and “Rhodococcus psychrotolerans” (98.2%) followed by Antrihabitans stalactiti (96.8%). However, the core gene-based phylogeny revealed that the novel isolate formed a tight cluster with A. stalactiti and was separated from R. cavernicola and other members of the family Nocardiaceae. The morphological and chemotaxonomic characteristics of strain YC3-6^T are in line with those of the genus Antrihabitans. Strain YC3-6^T showed an average nucleotide identity of 75.5% and a digital DDH of 20.3% with A. stalactiti. In addition, the core gene analysis showed that R. cavernicola formed a distinct subline between an Antrihabitans cluster and Aldersonia kunmingensis, and well separated from members of the genus Rhodococcus. The average amino acid identity values of R. cavernicola to closely related neighbours were 69.3–69.4% with members of the genus Antrihabitans and 67.3% with Ald. kunmingensis, while the POCP values ranged from 56.9 to 63.6%. On the basis of results obtained here, strain YC3-6^T is concluded to represent a novel species of the genus Antrihabitans, for which the name Antrihabitans stalagmiti sp. nov. (type strain, YC3-6^T?=?KACC 19963^T?=?DSM 107561^T) is proposed. Based on overall genome relatedness and chemotaxonomic differences, it is also proposed that R. cavernicola Lee et al. 2020 be transferred to a new genus Spelaeibacter as Spelaeibacter cavernicola gen. nov., comb. nov.

     

Morphological,Physiological, and Molecular Characterization of a Newly Isolated Steroid-Degrading Actinomycete,Identified as <Emphasis Type="Italic">Rhodococcus ruber</Emphasis> Strain Chol-4

The aerobic degradation of cholesterol, testosterone, androsterone, progesterone, and further steroid compounds as sole carbon source has been observed in the newly isolated bacterial Gram-positive strain Chol-4. The 16S rRNA gene sequence shares the greatest similarity with members of the genus Rhodococcus, with the closest shared nucleotide identities of 98–99% with Rhodococcus ruber (DSM 43338^T) and Rhodococcus aetherivorans (DSM 44752^T). Phylogenetic analysis of Rhodococcus 16S rRNA gene sequences consistently places strain Chol-4 in a clade shared with those both type strains within the Rhodococcus rhodochrous subclade. The results of DNA–DNA hybridization against its two phylogenetically closest neighbors as well as the results of morphological, physiological, and biochemical tests allowed genotypic and phenotypic differentiation of strain Chol-4 from Rhodococcus ruber (DSM 43338^T) on the species level and from the other validly described Rhodococcus species on the genus level. Strain Chol-4 therefore merits recognition as a novel strain of the species Rhodococcus ruber and demonstrates for the first time the capability of this species to utilize a great variety of steroid compounds as growth substrates never shown for other species of this genus so far. The genome of strain Chol-4 harbors at least one gene cluster that may be responsible for the degradation of steroid compounds. This gene cluster was identified in a cloned 5458 bp BamHI–EcoRV DNA fragment and compared to similar genes from other Gram-positive and Gram-negative bacteria described so far.     

Rhodococcus kronopolitis sp. nov., a novel actinobacterium isolated from a millipede (Kronopolites svenhedind Verhoeff)

A novel actinobacterium, designated strain NEAU-ML12^T, was isolated from a millipede (Kronopolites svenhedind Verhoeff), which was collected from Fenghuang Mountain in Wuchang, Heilongjiang Province, north China. The strain was characterized using a polyphasic approach. Strain NEAU-ML12^T was found to have morphological and chemotaxonomic characteristics typical of the members of the genus Rhodococcus. 16S rRNA gene sequence similarity analysis showed that the strain NEAU-ML12^T belongs to the genus Rhodococcus, and was most closely related to Rhodococcus tukisamuensis Mb8^T (98.9 %) and Rhodococcus koreensis DNP505^T (97.7 %). Phylogenetic analysis based on 16S rRNA gene sequences also demonstrated that strain NEAU-ML12^T should be classified in the genus Rhodococcus, forming a distinct clade with R. tukisamuensis Mb8^T supported by a 99 % bootstrap value. However, the DNA–DNA relatedness between strain NEAU-ML12^T and R. tukisamuensis Mb8^T was found to be 41.9 ± 0.7 %. Furthermore, strain NEAU-ML12^T could also be differentiated from R. tukisamuensis Mb8^T and other closely related strains (R. koreensis DNP505^T and Rhodococcus maanshanensis M712^T) by morphological and physiological characteristics. Therefore, it is proposed that strain NEAU-ML12^T represents a novel species of the genus Rhodococcus, for which the name Rhodococcus kronopolitis sp. nov. is proposed. The type strain is NEAU-ML12^T (=CGMCC 4.7145^T = DSM 46702^T).     

Degradation of 4-chlorobiphenyl and 4-chlorobenzoic acid by the strain <Emphasis Type="Italic">Rhodococcus ruber</Emphasis> P25

The strain Rhodococcus ruber P25 utilizes 4-chlorobiphenyl (4CB) and 4-chlorobenzoic acid (4CBA) as sole carbon and energy sources. 4CB degradation by washed cells of strain P25 was accompanied by transient formation of 4CBA, followed by its utilization and release of equimolar amounts of chloride ions into the medium. The strain R. ruber P25 possessed active enzyme systems providing 4CBA degradation via the stages of formation of intermediates, para-hydroxybenzoate (PHBA) and protocatechuic acid (PCA), to compounds of the basic metabolism. The involvement of protocatechuate 4,5-dioxygenase in 4CBA degradation by rhodococci was revealed. It was established that the initial stage of 4CBA degradation (dehalogenation) in the strain R. ruber P25 was controlled by the fcbA and fcbB genes encoding 4-CBA-CoA ligase and 4-CBA-CoA dehalogenase, respectively. The genes encoding 4CBA dehalogenase components have not been previously detected and characterized in bacteria of the genus Rhodococcus.     

A new family of Gonyleptoidea from South America (Opiliones,Laniatores)

A new family of Laniatores, Gerdesiidae fam. nov., is proposed based on molecular and morphological evidence. Data also indicate that this new family is the sister family of Tricommatidae. Gerdesiidae fam. nov. has a disjunct distribution, occurring in northern South America (Peru, Brazilian Amazon) and at a spot in south‐eastern Brazil (Minas Gerais State). The new family is composed of two genera: Gerdesius Roewer, 1952 (type genus) and G onycranaus gen. nov. (type species G onycranaus androgynus sp. nov. ). We propose the synonymy of Huralvioides H. Soares, 1970 with Gerdesius Roewer, 1952 based on molecular and morphological evidence. Three new species are described: G erdesius mapinguari sp. nov. (type locality: Brazil, Amazonas, Manaus, Reserva Km 41); G onycranaus androgynus sp. nov. (type locality: Brazil, Minas Gerais, Conceição do Mato Dentro); and an obligate cave‐dwelling species, G onycranaus pluto sp. nov. (type locality: Brazil, Minas Gerais, Morro do Pilar). © 2015 The Linnean Society of London     

Nocardia alba sp.nov., a novel actinomycete strain isolated from soil in China

A novel actinomycete strain, designated YIM 30243T, was isolated from a soil sample in Yunnan Province, China. Based on the results of phenotypic and genotypic characteristics, strain YIM 30243T should be assigned to a new species of the genus Nocardia, for which the name Nocardia alba sp. nov. is proposed. The type strain is YIM 30243T (= CCTCC AA001030T = DSM 44684T).     

<Emphasis Type="Italic">Nocardia zhihengii</Emphasis> sp. nov., an actinobacterium isolated from rhizosphere soil of <Emphasis Type="Italic">Psammosilene tunicoides</Emphasis>

A Nocardia-like actinobacterial strain, designated YIM TG2190^T, was isolated from rhizosphere soil of Psammosilene tunicoides collected from Gejiu, Yunnan province, China. The cells of strain YIM TG2190^T were observed to be Gram-stain positive and non-motile. The strain forms extensively branched substrate mycelia that fragments into rod-shaped elements. The 16S rRNA gene sequence analysis showed that strain YIM TG2190^T is closely related to Nocardia nova (97.5%), Nocardia jiangxiensis (97.1%) and Nocardia miyunensis (96.8%). Growth occurs at 4–30?°C (optimum 28?°C), pH 6.0–8.0 (optimum pH 7.0) and the strain can tolerate NaCl (w/v) up to 3% (optimum 0–1%). The cell walls were found to contain meso-diaminopimelic acid. The whole-cell sugars were identified as glucose, mannose, ribose, galactose, arabinose and fucose. The polar lipids were identified as diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannosides, phosphatidylglycerol and an unidentified phospholipid. The menaquinones detected were MK-9 (H₂) and MK-8 (H₄). The major fatty acids (>?5%) were found to be C_16:0 (33.9%), summed feature 3 (21.7%), C_18:0 10-methyl TBSA (13.7%) and C_18:1ω9c (7.0%). The DNA G+C content was determined to be 61.1 mol%. DNA-DNA relatedness between the strain YIM TG2190^T and N. nova CGMCC 4.1705^T, N. jiangxiensis CGMCC 4.1905^T and N. miyunensis CGMCC 4.1904^T were 46.9?±?2.6, 36.8?±?1.3, and 35.7?±?2.6%, respectively, values which are less than the threshold value (70%) for the delineation of prokaryotic genomic species. The phenotypic, chemotaxonomic and phylogenetic data indicates that strain YIM TG2190^T represents a novel species of the genus Nocardia, for which the name Nocardia zhihengii sp. nov. is proposed. The type strain is YIM TG2190^T (=KCTC 39596^T?=?DSM 100515^T).