Parapseudoflavitalea muciniphila gen. nov., sp. nov., a member of the family Chitinophagaceae isolated from a human peritoneal tumour and reclassification of Pseudobacter ginsenosidimutans as Pseudoflavitalea ginsenosidimutans comb. nov.

Abstract

A Gram-stain-negative, microaerophilic, non-motile, rod-shaped bacterium strain designated PMP191F^T, was isolated from a human peritoneal tumour. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the organism formed a lineage within the family Chitinophagaceae that was distinct from members of the genus Pseudoflavitalea (95.1–95.2 % sequence similarity) and Pseudobacter ginsenosidimutans (94.4 % sequence similarity). The average nucleotide identity values between strain PMP191F^T and Pseudoflavitalea rhizosphaerae T16R-265^T and Pseudobacter ginsenosidimutans Gsoil 221^T was 68.9 and 62.3% respectively. The only respiratory quinone of strain PMP191F^T was MK-7 and the major fatty acids were iso-C_15 : 0, iso-C_15 : 1 G and summed feature 3 (C_16:1 ω7c and/or C_16:1 ω6c). The polar lipids consisted of phosphatidylethanolamine and some unidentified amino and glycolipids. The G+C content of strain PMP191F^T calculated from the genome sequence was 43.4 mol%. Based on phylogenetic, phenotypic and chemotaxonomic evidence, strain PMP191F^T represents a novel species and genus for which the name Parapseudoflavitalea muciniphila gen. nov., sp. nov. is proposed. The type strain is PMP191F^T (=DSM 104999^T=ATCC BAA-2857^T = CCUG 72691^T). The phylogenetic analyses also revealed that Pseudobacter ginsenosidimutans shared over 98 % sequence similarly to members of the genus Pseudoflavitalea . However, the average nucleotide identity value between Pseudoflavitalea rhizosphaerae T16R-265^T, the type species of the genus and Pseudobacter ginsenosidimutans Gsoil 221^T was 86.8 %. Therefore, we also propose that Pseudobacter ginsenosidimutans be reclassified as Pseudoflavitalea ginsenosidimutans comb. nov.

The family Chitinophagaceae was created by Kämpfer et al., mostly on the basis 16S rRNA gene sequences of a number of Gram-negative staining, aerobic or facultatively anerobic organisms [1] and currently consists of 38 genera with validly published names (https://lpsn.dsmz.de/family/chitinophagaceae). Members of this family are widely distributed with the majority isolated from soil, sediment and water of lakes as well as seawater [1, 2]. The genus Pseudoflavitalea was proposed by Kim et al. to accommodate an organism recovered from the rhizosphere of tomato which was Gram-stain-negative, aerobic and rod-shaped and possessed MK-7 as the respiratory quinone and the major polar lipid was phoshatidylethanolamine and an unknown lipid [3]. The predominant fatty acids consisted of iso-C_15 : 0, iso-C_15 : 1 G and iso-C_17 : 0 3-OH. The authors also found that Flavitalea soli , isolated from soil was found to be phylogenetically related to Pseudoflavitalea and possessed phenotypic and chemotaxonomic traits that were consistent with Pseudoflavitalea and therefore was reclassified into this novel genus [3].

Pseudomyxoma peritonei (PMP) is a clinical syndrome where mucus-secreting tumours are disseminated throughout the peritoneal cavity [4]. In a continuing study into the microbial communities that make up the microbiome of such a tumour, a Gram-stain-negative, microaerobic, non-motile and rod-shaped bacterium that resembled members of the genus Pseudoflavitalea was isolated [5]. Based on the information obtained from a polyphasic taxonomic approach, it is proposed that strain PMP191F^T represents a novel species and genus for which the name Parapseudoflavitalea gen., nov. sp. nov. is proposed. Furthermore, it is proposed that Pseudobacter ginsenosidimutans be reclassified as Pseudoflavitalea ginsenosidimutans comb. nov.

Isolation and ecology

Samples were obtained during a study into the microbiome associated with PMP tumours, a rare form of peritoneal cancer that originates in the appendix [4]. The samples were collected under an institutional review board (IRB)-approved protocol at Mercy Medical Center, Baltimore, MD, USA, in which patients underwent cytoreductive surgery to remove all visible tumour nodules from the peritoneal cavity and from the surfaces of abdominal organs. All patients signed informed consent forms prior to participation. Isolate PMP191F^T was recovered from a female patient with a diagnosis of disseminated peritoneal adenomucinosis, a subtype of PMP. Isolates were initially recovered using Ham’s F-12 (Sigma) supplemented with 2 % foetal bovine serum. For routine maintenance, strain PMP191F^T was grown on Campylobacter Blood Agar (Hardy Diagnostics) supplemented with 10 % sheep blood.

16S rRNA gene phylogeny

The nearest known relatives for strain PMP191F^T were determined using the EzBioCloud-server (http://eztaxon-e.ezbiocloud.net/ [6]. The sequences were retrieved and aligned with strain PMP191F^T using Clustal W [7] and the phylogenetic analysis was performed using mega 7 [8] using the maximum-likelihood [9] and neighbor-joining methods [10] employing the Kimura two-parameter substitution model [11]. Bootstrap analysis with 1000 replicates were used to determine support for the tree topology [12]. The phylogenetic analyses demonstrated that strain PMP191F^T belonged within the family Chitinophagaceae , class Sphingobacteria , phylum Bacteroidetes . The highest sequence similarities were found to Pseudoflavitalea soli (95.2 %), Pseudoflavitalea rhizosphaerae (95.1 %) and Pseudobacter ginsenosidimutans (94.4 %); the phylogenetic tree demonstrates that Pseudoflavitalea soli , Pseudoflavitalea rhizosphaerae and Psudobacter ginsenosidimutans form a cluster that is supported by high bootstrap values (Fig. 1). Strain PMP191F^T represents a distinct lineage at the base of the cluster with Pseudoflavitala and Pseudobacter but this relationship is not supported by a significant bootstrap value (Fig. 1). The almost identical tree topology was obtained using the neighbor-joining method (Fig. S2, available in the online version of this article). The analyses demonstrated the close association of Pseudobacter ginsenosidimutans with Pseudoflavitalea soli (98.1 %), Pseudoflavitalea rhizosphaerae (98.5 %) that confirms the recent findings of Zhang et al. [13]. These values are below the value (98.8 %) recommended for delineation of species but higher than the value (94.5 %) routinely used for the separation of genera based on 16S rRNA gene sequence analysis [14]. On examination of the literature, members of Pseudoflavitalea were not included in the phylogenetic analysis with Pseudobacter ginsenosidimutans [15].

Fig. 1.

Phylogenetic tree of 16S rRNA gene sequences indicating the position of strain PMP191F^T with some representative members of the family Chitinophagaceae . The tree was reconstructed using the maximum-likelihood algorithm from mega (version 7) with Flavobacterium aquatile as the outgroup. Bootstrap values (%) were obtained with 1000 replicates and are displayed on their relative branches. Bar, 1 %.

Genome features

The entire genome of PMP191F^T was sequenced as reported previously [16], in brief PMP191F^T genomic DNA was subjected to DNA library construction using the Illumina MiSeq platform and sequenced with a 2×250 bp paired-end sequencing mode [17]. A total of 17.8 million quality reads of 225 bp average length were assembled into 66 contigs (N50, 269 kbp) using the CLC Genomics Workbench (version 6.0; CLC bio). These contigs yielded a total assembly size of 6 452 579 bp. Functional annotation was performed with Rapid Annotations using Subsystems Technology (rast) [18], which predicted 5577 coding regions with 380 subsystems and 46 tRNAs. Several genes were predicted to impart antibiotic resistance, including β-lactamase and multiple efflux pumps. Furthermore, it lacks most of the genes for flagella and twitching motility that supports the observation that the organism is non-motile.

Due to the high 16S similarity values, the average nucleotide identity (ANI) values were determined by the online calculator https://www.ezbiocloud.net/tools/ani [19] and the average amino acid identity (AAI) values [20] were also calculated using Prodigal [21] and the Enevomics suite of programmes [22]. The values between strain PMP191F^T (LGYU00000000) and the available genomes for Pseudoflavitalea rhizosphaerae T16R-265^T (NZ_RCSU00000000) and Pseudobacter ginsenosidimutans Gsoil 221^T (NZ_CP042431) were 68.9 and 62.3%, respectively, which are well below the 95–96 % cutoff value routinely used for the delineation of species [19]. For the AAI values between strain PMP191F^T (LGYU00000000) and the available genomes for Pseudoflavitalea rhizosphaerae T16R-265^T (NZ_RCSU00000000) and Pseudobacter ginsenosidimutans Gsoil 221^T (NZ_CP042431) were 63.4 and 63.1%, respectively [18]. The ANI/AAI values between Pseudoflavitalea rhizosphaerae T16R-265^T and Pseudobacter ginsenosidimutans Gsoil 221^T were 86.8 and 91.7% respectively. It has been demonstrated that AAI values of 74–76 % can be used to delineate species of the same genus [23] .Although the 16S rRNA gene similarity value between Pseudoflavitalea rhizosphaerae T16R-265^T and Pseudobacter ginsenosidimutans Gsoil 221^T was 98.5 % and is very close to values routinely used for the delineation of species [14] the ANI value of 86.8 % and AAI value of 91.7 % demonstrates that indeed it represents a distinct species of the genus Pseudoflavitalea but not a separate genus ( Pseudobacter ) as originally described [15].

Physiology and chemotaxonomy

The cell morphology of strain PMP191F^T was observed using a phase-contrast microscope (Olympus CX41) at ×1000 magnification after cells were grown for 2 days at 37 °C on Reasoner’s 2A agar medium (made using R2A broth media, Teknova R005) and using a Jeol JSM-IT100 scanning electron microscope following growth in Ham’s F12 medium, fixation with 2 % glutaraldehyde, and sputter coating with gold (Fig. S1). The physiological and biochemical characteristics of the isolate were determined using a combination of conventional tests as described previously [24]. Physiological characteristics were determined at varying temperature growth ranges (5.0–60 °C, in increments of 5 °C) and pH values (5.0–9.5, in increments of 0.5 pH units). For the different pH ranges, the following buffers were prepared at a final concentration of 10 mM with the following; sodium acetate (pH 5.0), potassium phosphate (6.0–8.0), Tris-HCl (pH 7.0–9.0), and sodium borate (pH 9.5). In addition, growth at pH 10 and 11 was measured using carbonate/bicarbonate buffer as described previously [3]. Salt tolerance was examined using different concentrations of NaCl (0.0, 0.5, 1.0–9.0 % (w/v) in increments of 1.0 % for concentrations above 1 %). Optimal growth conditions were determined by monitoring the optical density using a spectrophotometer at 600 nm (Spectronic 20D, Milton Roy). Growth resulting in an increase of OD_600nm of >0.1 was considered to indicate growth. All tests were performed in duplicate. Oxygen tolerance was tested using anaerobic jars, tri-gas variable atmosphere incubators, and standard incubators. The conditions tested were anaerobic, 5 % O₂+10 % CO₂, 10 % CO₂ (~17.5 % O₂), 5 % CO₂ (~18.5 % O₂) and atmospheric oxygen (~21 % O₂). All tests on strain PMP191F^T, Pseudoflavitalea soli (KACC 17319^T) and Pseudoflavitalea rhizosphaerae (KACC 18655^T) were performed in parallel. When compared to the soil isolates, strain PMP191F^T appeared to be quite unreactive for single carbon assimilation studies, only weakly utilising maltose, mannose and trehalose whereas Pseudoflavitalea rhizosphaerae (KACC 18655^T), utilised glucose, trehalose, mannose and lactate and Pseudoflavitalea soli (KACC 17319^T) utilised glucose, mannose, N-acetyl-glucosamine, maltose, lactate and trehalose (weak). Additional biochemical characterizations were carried out using the GEN III Microplate system (Biolog), API ZYM and API 20NE test systems (bioMérieux) following the manufacturer’s instructions. The results of these physiological tests are given in Tables 1 and S1 and the species description.

Table 1.

Cellular fatty acid compositions of strain PMP191F^T and its closest relatives

Strains: 1, PMP191F^T (this study); 2, Pseudoflavitalea rhizosphaerae T16R-265^T (this study); 3, Pseudoflavitalea soli KACC 17319^T (this study); 5, Pseudobacter ginsenosidimutans Gsoil 221^T [15]. Values are percentages of total fatty acids; fatty acids amounting to <1 % of total fatty acids in all strains studied are not listed. Major components are indicated in bold type.

Fatty acid	1	2	3	4
iso-C13 : 0	–	–	1.1	–
iso-C13 : 0 3-OH	–	–	–	–
C14 : 0	–	–	–	1.0
C15 : 0 2-OH	–	1.5	1.0	–
C15 : 0 3-OH	–	–	1.0	–
iso-C15 : 0 3-OH	–	–	3.1	2.0
anteiso-C15 : 0	2.0	3.6	3.2	4.3
iso-C15 : 0	34.7	33.3	27.9	35.8
anteiso-C15 : 1 A	1.1	1.2	0.8	–
iso-C15 : 1 G	20.1	16.1	14.6	10.6
C15 : 1ω5c	4.4
C16 : 0	7.6	2.5	1.5	4.3
C16 : 0 3-OH	1.3	2.7	2.8	4.1
iso-C16 : 0 3-OH	–	–	1.6	2.0
iso-C16 : 0	–	–	–	1.6
C17 : 0 2-OH	–	1.8	1.6	2.1
C17 : 0 3-OH	–	1.1	1.6
iso-C17 : 0 3-OH	6.1	24.3	26.2	24.0
Unknown 13.565	–	1.2	–	–
Unknown 16.582	–	1.6	1.8	–
Summed feature 3	13.0	5.8	7.7	3.7

For chemotaxonomic investigations, fatty acids and polar lipids analyses of PMP191F^T, Pseudoflavitalea soli (KACC 17319^T) and Pseudoflavitalea rhizosphaerae (KACC 18655^T) were performed at the Center for Microbial Identification and Taxonomy (CMIT; University of Oklahoma, Norman, Oklahoma). Cells were grown on R2A agar medium at 28 °C for 2 days, conditions were chosen to allow comparison with values available in the literature. Fatty acid methyl esters (FAMEs) were extracted using the Sherlock Microbial Identification System (midi, Inc.) version 6.1 as described previously [25, 26]. Analysis was carried out with an Agilent Technologies 6890 N gas chromatograph equipped with a phenyl methyl silicone fused silica capillary column (HP-2 25m×0.2 mm×0.33 μm film thickness) and a flame ionisation detector. Hydrogen was used as the carrier gas. The temperature programme was initiated at 170 °C and increased at 5 °C min⁻¹ to a final temperature of 270 °C. Using the RTSBA6 library, the relative amount of each fatty acid was expressed in terms of the percentage of total fatty acids. The major fatty acids were iso-C_15 : 0 (34.7 %), iso-C_15 : 1 G (20.1 %) and components contained in summed feature 3 (C_15 : 0 2-OH and/or C_16 : 1 ω7c; 13.0 %, a complete profile is shown in Table 1. iso-C_15 : 0 and iso-C_15 : 1 G are produced by strain PMP191F^T, P. rhizosphaerae and P. soli , this product is also reported from P. ginsenosidimutans grown under similar conditions [15]. However, the novel organism can be differentiated from its near relatives by failing to produce major amounts iso-C_17 : 0 3-OH and in producing major products contained in summed feature 3 (C_15 : 0 2-OH and/or C_16 : 1 ω7c). Species of the genus Pseudoflavitalea are recovered from soil, whereas strain PMP191F^T was isolated from human clinical material and has a higher optimum growth temperature than the former; therefore, the fatty acid profile was also determined at 37 °C (Table S2). This is important so this information is available for the comparison of related organisms isolated at some point in the future. As expected, some changes in the profile occurred from that obtained at 28 °C. The major product remained as iso-C_15 : 0 but iso-C_15 : 1 G fell below 10 % and C_15:1ω5c was increased to constitute a major product (12.6 %). Furthermore, summed feature 3 (C _15 : 0 2-OH and/or C_16 : 1 ω7c) was no longer produced as a major product (7.4 %).

Polar lipids were determined for PMP191F^T, Pseudoflavitalea soli (KACC 17319^T) and Pseudoflavitalea rhizosphaerae (KACC 18655^T) according to Tindall [27]. The only dominant polar lipid for all three strains was PE (phosphatidylethanolamine) (Fig. S3) which is consistent with published results for both P. rhizosphaerae and P. soli [3]. P. ginsenosidimutans differs in producing diphosphatidylglycerol (DPG) as a major product in addition to PE [15]. The genome is playing an increasing role in taxonomy, the application of in-silico analysis is now being introduced to replace characteristics traditionally determined in the laboratory with the dual goals of increasing the speed of the description of taxa and the accuracy and consistency of characteristics reported [28–30]. For example, laboratory-based DNA–DNA hybridization methods have now been routinely replaced by ANI and in silico DNA–DNA analysis [31–33]. Phenotypic and chemotaxonomic traits derived from genome-based studies have also been useful in taxonomic identification and classification of bacteria [34, 35]. In this study we have applied this approach to polar lipid analysis and used the KEGG database to investigate glycerophospholipid metabolism [36, 37]. The results indicate that strain PMP191F^T contains phosphatidylserine decarboxylase (psd, E.C. 4.1.1.65) that normally, and very efficiently, catalyses the production of PE from phosphatidyl-l-serine (Figs S4 and S5) [38, 39]. However, the intermediate phosphatidyl-l-serine is rarely seen on TLC plates and we hypothesize that this depends largely on the age of the cells when harvested for laboratory analysis and the fact this is an intermediate used in the formation of PE. It is pertinent to note that the enzyme phosphatidylserine synthase (E.C. 2.7.8.8, pssA) used in the synthesis of phosphatidylserine is not identified utilising the KEGG database with strain PMP191F^T. We have reported this discrepancy in our previous studies [37] and may be due to an alternative pathway in the synthesis of phosphatidylserine. This information is consistent with the results obtained from the laboratory-determined products (PE) observed on the TLC plate (Fig. S3). In addition, a number of enzymes involved in biosynthesis of other lipids are also present in the genome of PMP191F^T are likely to correspond to unidentified products seen on the TLC plate (Fig. S3). The same approach demonstrated that P. ginsenosidimutans Gsoil 221^T also contained the genes required for the synthesis of PE and DPG (Fig. S6) that was consistent with the previously reported results [15]. Isoprenoid quinones were extracted and analysed by HPLC according to previously described methods [40, 41]. The menaqinone was MK-7 which is also reported for the genera Pseudoflavitalea , Pseudobacter and other members of the family Chitinophagaceae [1]. The G+C content was calculated from whole genome sequencing and was determined to be 43.4 mol%.

Characteristics that serve to differentiate the novel organism from its close relatives are shown in Tables 1 and 2. Although the 16S rRNA gene similarly values for strain PMP191F^T and members of the genus Pseudoflavitalea are on the borderline of values used to delineate novel genera [14]; the topology of the phylogenetic tree demonstrating that strain PMP191^T forming a distinct lineage strongly supports the separateness of the novel organism from Pseudoflavitalea species and it represents a novel genus. Furthermore, members of the genus Pseudoflavitalea and P. ginsenosidimutans are recovered from soil whereas strain PMP191F^T was isolated from human clinical material. These different ecosystems will play a significant role in shaping the physiological properties and metabolic capabilities of the organisms contributing to evolutionary divergence between organisms [42, 43].

Table 2.

Phenotypic differences among strain PMP 191^T and its closest relatives

Taxa: 1, Parapseudoflavitalea muciniphila (this study); 2, Pseudoflavitalea rhizosphaerae (this study and [3]; 3. Pseudoflavitalea soli (this study and [3] Pseudobacer ginsenosidimutans [15]. w, Weak reaction; nr, not reported

Characteristic	1	2	3	4
Cell shape	Rod	Rod or long rod	Rod	Peudo-rod
Colony colour	Orange-red	Light yellow	Yellow	Milky
Temperature range for growth (°C)	20–45	10–40	10–37	10–37
pH range for growth	5–8	6–9	5–10	5–8
NaCl (% w/v) range for growth	<0.5–1.0	0.5–1.0	<0.5	0.5–1.0
API ZYM tests:
N-Acetyl-β-glucosaminidase	+	+	+	−
Acid phosphatase	+	w	+	+
α-Chymotrypsin	+	−	−	+
Cystine arylamidase	+	w	−	+
Esterase C4	−	−	−	+
Esterase lipase C8	−	−	−	+
β-Glucuronidase	+	−	−	−
α-Mannosidase	−	−	−	+
Trypsin	+	−	−	+
API 20NE tests:
Gelatinase	−	+	−	+
d-mannitol	−	−	−	+
l-Arabinose	+	w	−	−
Assimilation of:
Maltose	w	−	+	+
Glucose	−	+	+	−
Gluconate	−	+	−	nr
Trehalose	w	+	−	nr
Lactate	−	+	+	nr
DNA G+C (mol%)	43.4	46.2	55.7	47.1
Source	Human peritoneal tumour	Rhizosphere of tomato	Soil	Soil

On the basis of the data obtained from our study using a polyphasic taxonomic that included phenotypic and phylogenetic information, strain PMP191F^T merits recognition as representing a novel species of a novel genus for which the name Parapseudoflavitalea muciniphila gen. nov., sp. nov. is proposed. Furthermore, phylogenetic analyses, biochemical and chemotaxonomic characteristics of P. ginsenosidimutans are consistent with members of the genus Pseudoflavitalea and therefore we propose to reclassify P. ginsenosidimutans as Pseudoflavitalea ginsenosidimutans comb. nov.

Description of Parapseudoflavitalea gen. nov.

Parapseudoflavitalea GEN. NOV (Pa.ra.pseu.do.fla.vi.ta’le.a. Gr. prep. para beside, near, like; N.L. fem. n. Pseudoflavitalea a bacterial genus name, N.L. fem. n. Parapseudoflavitalea beside Pseudoflavitalea , referring to the similarity of the two genera).

Cells are Gram-stain-negative, microaerophilic, non-motile and rod-shaped. The only respiratory quinone is MK-7. At 28 °C and the major fatty acids are iso-C_15 : 0, iso-C_15 : 1 G and summed feature 3 (C_16:1 ω7c and/or C_16:1 ω6c), but at 37 °C the major products are iso-C_15 : 0 and C_15:1 ω5c. The polar lipids consisted of PE and some unidentified amino and glycolipids. The G+C content of the type species is 43.4 mol%. Based on 16S rRNA sequence analysis, the genus is a member of the family Chitinophagaceae . The type species is Parapseudoflavitalea muciniphila .

Description of Parapseudoflavitalea muciniphila sp. nov.

Parapseudoflavitalea muciniphila SP. NOV (mu.ci.ni’phi.la. N.L. neut. n. mucinum mucin; Gr. adj. philos loving; N.L. fem. adj. muciniphila mucin-loving).

Cells are medium to very long rods (average length 3.4 µm; range, 1.3–9.8 µm). When grown on R2A or blood agar, colonies are round, smooth, convex and orange-red in colour. Grows at 10–37 °C. Tolerates up to 1 % NaCl. The organism fails to grow under anaerobic or atmospheric oxygen conditions on agar plates or in broth. Growth is optimal in a 5 % CO₂ incubator, corresponding to ~18.5 % O_2. Growth was observed at pH values ranging from pH 5–8. Nitrate is not reduced to nitrite. Oxidase positive, catalase activity is very weak. Negative for indole production. Aesculin is hydrolysed. Using the single carbon plate assimilation test, maltose, mannose and trehalose are weakly utilised but glucose, arginine, norleucine, arabinose, mannitol, N-acetyl-glucosamine, gluconate, caprate, adipate, malate, citrate, phenylactate, sucrose and lactate were not. Using the Biolog GEN III MicroPlate system, positive reactions are produced from glucose, mannose, N-acetylglucosamine, dextrin, maltose, trehalose, cellobiose, gentiobiose, sucrose, lactose, melibiose, methyl β-d-glucoside, salicin, N-acetyl-β-d-mannosamine, d-galactose, l-rhamnose, ritamycin SV, gelatin, glycyl-l-proline, aspartic acid, lincomycin, pectin, d-galacturonic acid, glucuronic acid, tetrazolium blue, methyl pyruvate, d-lactic acid methyl ester, Tween 40 and aztreonam. Using the API ZYM test system, positive reactions are obtained with N-acetyl-β-glucosaminidase, acid phosphatase, alkaline phosphatase, α-chymotrypsin, cystine arylamidase, α-galactosidase, β-galactosidase, β-glucuronidase (weak), α-glucosidase, β-glucosidase, leucine arylamidase, lipase C₁₄ (weak), naphthol-AS-BI-phosphohydrolase, trypsin and valine arylamidase. According to the API 20NE test system, positive reactions are obtained with l-arabinose, aesculin, d-glucose, maltose, d-mannose, oxidase and PNPG-β-galactosidase.The major fatty acids are iso-C_15 : 0, iso-C_15 : 1 G, summed feature 3 (C_15 : 0 2-OH and/or C_16:1ω7c). The only quinone is MK-7. The predominant polar lipid is PE, with a number of unidentified products that consist of five phospholipids, two amino-containing lipids and three other lipids.

The type strain, PMP191F^T (=DSM 104999^T=ATCC BAA-2857^T =CCUG 72691^T), was isolated from a human peritoneal tumour. The DNA G+C content of the type strain is 43.4 % (Genome).

Description of Pseudoflavitalea ginsenosidimutans comb. nov.

Pseudoflavitalea ginsenosidimutans COMB. NOV (gin.se.no.si.di.mu’tans. N.L. neut. n. ginsenosidum ginsenoside; L. pres. part. mutans transforming, converting; N.L. part. adj. ginsenosidimutans ginsenoside-converting).

Basonym: Pseudobacter ginsenosidimutans Siddiqi and Im 2016.

The description is identical to that proposed for Pseudobacter ginsenosidimutans.

Source: Soil ginseng field.

The type strain is Gsoil 221^T (=KACC 14278^T=DSM 18116^T). The G+C content of the DNA of the type strain is 47.1 mol% (HPLC).

Emended description of the genus Pseudoflavitalea

The genus description is as given by Kim et al. (2016) with the addition of the following, While PE are observed to be the predominant polar lipid, minor amounts of DPG, PG and phosphatidylmethylethanolamine may also be produced.