Mumia flava gen. nov., sp. nov., an actinobacterium of the family Nocardioidaceae
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International Journal of Systematic and Evolutionary Microbiology (2014), 64, 1461–1467 DOI 10.1099/ijs.0.058701-0
Mumia flava gen. nov., sp. nov., an actinobacterium
of the family Nocardioidaceae
Learn-Han Lee,1 Nurullhudda Zainal,1,2 Adzzie-Shazleen Azman,1
Nurul-Syakima Ab Mutalib,3 Kui Hong4 and Kok-Gan Chan2
Correspondence 1
Jeffrey Cheah School of Medicine and Health Sciences, Monash University Sunway Campus,
Learn-Han Lee 46150 Bandar Sunway, Selangor Darul Ehsan, Malaysia
lee.learn.han@monash.edu or 2
Division of Genetics and Molecular Biology, Institute of Biological Sciences, Faculty of Science,
leelearnhan@yahoo.com University of Malaya, 50603 Kuala Lumpur, Malaysia
3
UKM Medical Molecular Biology Institute (UMBI), UKM Medical Centre, Bandar Tun Razak,
56000 Cheras, Kuala Lumpur, Malaysia
4
Key Laboratory of Combinatory Biosynthesis and Drug Discovery, Ministry of Education,
Wuhan University, School of Pharmaceutical Sciences, Wuhan, PR China
A novel actinobacterial strain, designated MUSC 201T, was isolated from a mangrove soil
collected from Kuantan, the capital city of Pahang State in Malaysia. The taxonomic status of this
strain was determined using a polyphasic approach. Comparative 16S rRNA gene sequence
analysis revealed that strain MUSC 201T represented a novel lineage within the class
Actinobacteria. Strain MUSC 201T formed a distinct clade in the family Nocardioidaceae and was
most closely related to the members of the genera Nocardioides (16S rRNA gene sequence
similarity, 91.9–95.1 %), Aeromicrobium (92.7–94.6 %), Marmoricola (92.5–93.1 %) and
Kribbella (91.5–92.4 %). The cells of this strain were irregular coccoid to short rod shaped. The
peptidoglycan contained LL-diaminopimelic acid as diagnostic diamino acid and the peptidogly-
can type was A3c. The peptidoglycan cell wall contained LL-diaminopimelic acid, glycine, glutamic
acid and alanine in a molar ratio of 1.5 : 0.9 : 1.0 : 1.5. The cell-wall sugars were galactose and
rhamnose. The predominant menaquinone was MK-9(H4). The polar lipids consisted of
diphosphatidylglycerol, phosphatidylglycerol, phosphoglycolipid, glycolipid and four unknown
phospholipids. The major cellular fatty acids were C18 : 1v9c (30.8 %), C16 : 0 (24.1 %), and 10-
methyl C18 : 0 (13.9 %). The DNA G+C content was 72.0±0.1 mol%. On the basis of
phylogenetic and phenotypic differences from members of the genera of the family
Nocardioidaceae, a novel genus and species, Mumia flava gen. nov., sp. nov. are proposed. The
type strain of Mumia flava is MUSC 201T (5DSM 27763T5MCCC 1A00646T5NBRC
109973T).
The family Nocardioidaceae was first proposed by Nesterenko belonging to the family Nocardioidaceae, designated
et al. (1985) and the name was validly published in 1990 MUSC 201T, which was isolated from a mangrove soil
(Nesterenko et al., 1990). The description of the family was sample collected from Kuantan, the capital city of Pahang
revised by Zhi et al. (2009). At the time of writing, the family State, Peninsular of Malaysia. In order to determine the
Nocardioidaceae comprises seven genera: Nocardioides (Prauser, taxonomic and phylogenetic position of strain MUSC 201T,
1976), Aeromicrobium (Miller et al., 1991), Kribbella (Park the morphology, physiological and biochemical character-
et al., 1999; Sohn et al., 2003), Marmoricola (Urzı̀ et al., 2000), istics, chemotaxonomic markers and 16S rRNA gene
Actinopolymorpha (Wang et al., 2001), Thermasporomyces sequence of the novel strain were examined and analysed.
(Yabe et al., 2011) and Flindersiella (Kaewkla & Franco, The results indicated that strain MUSC 201T represented a
2011). The present investigation was designed to determine novel species of a new genus, for which the name Mumia
the taxonomic status of a novel actinobacterial strain flava gen. nov., sp. nov. is proposed.
The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene A soil sample was collected in December 2012. Topsoil
sequence of strain MUSC 201T is KC907394. samples of the upper 20 cm layer (after removing the top
One supplementary figure and one supplementary table are available 2–3 cm) were collected and sampled into sterile plastic
with the online version of this paper. bags using an aseptic metal trowel, and stored at 220 uC.
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Five grams of air-dried soil sas mix with 45 ml sterilized sugars of strain MUSC 201T was carried out by the Iden-
water and mill-ground, then spread onto selective isolation tification Service of the DSMZ, Braunschweig, Germany.
medium: yeast malt agar [International Streptomyces Project The analyses were carried out according to published pro-
(ISP) 2 medium; Shirling & Gottlieb, 1966] supplemented tocols (Schumann, 2011). Major diagnostic whole-organism
with cycloheximide (25 mg ml21) and nystatin (10 mg ml21) sugars of strain MUSC 201T were obtained following a
and incubated at 28 uC for 7 days. The strain MUSC 201T procedure described by Whiton et al. (1985) and analysed by
was maintained on ISP2 medium at 28 uC and as glycerol TLC on cellulose plates according to Staneck & Roberts
suspensions (20 %, v/v) at 220 uC. (1974). Analysis of respiratory menaquinones and polar
lipids was carried out by the Identification Service of the
Cultural characteristics of strain MUSC 201T were deter-
DSMZ. The cellular polar lipids were extracted and analysed
mined following growth on ISP2 and ISP 7 media (Shirling
by TLC (Kates, 1986). Cellular fatty acid analysis of strain
& Gottlieb, 1966), starch casein agar (SCA; Küster &
MUSC 201T and closely related type strains was carried out
Williams 1964), Streptomyces agar (SA; Atlas 1993), actino-
by the Identification Service of the DSMZ. The cell mass of
mycete isolation agar (AIA; Atlas 1993) and nutrient agar
strain MUSC 201T was harvested from TSB after incubation
(MacFaddin, 2000) for 7 days at 28 uC. The ISCC-NBS
at 28 uC for 5 days. The fatty acids were extracted and
colour charts were used to determine the names and
prepared according to the standard protocol of the MIDI
designations of colony colours (Kelly, 1964). Light micro-
(Microbial Identification) system (Sasser, 1990).
scopy (80i, Nikon) and scanning electron microscopy (JSM
6400, JEOL) were used to observe the morphologies of Genomic DNA extractions, PCR amplification and sequen-
strains after incubation on ISP2 medium at 28 uC for 7 days. cing of the 16S rRNA gene of strain MUSC 201T were
Gram staining was performed by the standard Gram carried out as described by Hong et al. (2009). The 16S
reaction and was confirmed by using KOH lysis (Cerny, rRNA gene sequence of strain MUSC 201T was aligned
1978). The growth temperature was tested at 12–52 uC at manually with sequences from the most closely related
intervals of 4 uC on ISP2 medium. NaCl tolerance was tested genera classified in the family Nocardioidaceae that had
using tryptic soy broth (TSB) (casein, 17 g; soybean meal, 3 been retrieved from the GenBank/EMBL/DDBJ databases
g; dextrose, 2.5 g; dipotassium hydrogen phosphate, 2.5 g; using CLUSTAL X software (Thompson et al., 1997). The
distilled water, 1 L; pH 7.3) and salt concentrations ranging alignment was manually verified and adjusted prior to the
from 0–18 % (w/v) at intervals of 2 %. The pH range for reconstruction of a phylogenetic tree. Phylogenetic trees were
growth was tested between pH 4.0 and 10.0 at intervals of reconstructed using the neighbour-joining (Saitou & Nei,
1 pH unit. Carbon-source utilization and chemical sensitiv- 1987), maximum-likelihood (Felsenstein, 1981) and max-
ity assays were determined using Biolog GenIII MicroPlates imum-parsimony (Fitch, 1971) algorithms using MEGA
according to the manufacturer’s instructions. Catalase version 5.2 (Tamura et al., 2011). Calculations of sequence
activity was determined by bubble production in a 3 % (v/ similarity level were carried out using the EzTaxon-e server
v) hydrogen peroxide solution. Production of melanoid (http://eztaxon-e.ezbiocloud.net/; Kim et al., 2012). The
pigments was examined using tyrosine agar (ISP 7). stability of the resultant tree topologies was evaluated by
Haemolytic activity tests were performed in blood agar using the bootstrap resampling method of Felsenstein (1985).
medium containing 5 % (w/v) peptone, 3 % (w/v) yeast Evolutionary distances were computed using Kimura’s two-
extract, 5 % (w/v) NaCl and 5 % (v/v) human blood parameter model (Kimura, 1980). Terrabacter tumescens
(Carrillo et al., 1996). Plates were examined for haemolysis DSM 20308T was used as an outgroup. The genomic DNA
after incubation at 32 uC for 5 days. Presence of a clear zone of strain MUSC 201T for the determination of G+C content
around colonies signified the potential of isolates for sur- was extracted according to the method of Cashion et al.
factant production. Lipase, amylase, cellulase, chitinase, (1977). The G+C content of the DNA was determined by
protease and xylanase activities were determined by growing HPLC (Mesbah et al., 1989).
cells on ISP 2 medium and following protocols as described
An almost complete 16S rRNA gene sequence was
by Meena et al. (2013). Antibiotic susceptibility tests were
determined for strain MUSC 201T (1486 bp). A phylogen-
performed by the disc diffusion method as described by Shieh
etic tree was reconstructed based on the 16S rRNA gene
et al. (2003). Antimicrobials used and their concentrations
sequences (Fig. 1). The comparative 16S rRNA gene
were as follows: ampicillin (10 mg), ampicillin sulbactam
sequence analysis showed that strain MUSC 201T fell
(30 mg), cefotaxime (30 mg), cefuroxime (30 mg), cephalos-
within the evolutionary radiation occupied by the family
porin (30 mg), chloramphenicol (30 mg), ciprofloxacin
Nocardioidaceae (Fig. 1). The closest phylogenetic neigh-
(10 mg), erythromycin (15 mg), gentamicin (20 mg), nalidixic
bours were members of the genera of the family
acid (30 mg), penicillin G (10 mg), streptomycin (10 mg),
Nocardioidaceae. Strain MUSC 201T showed 16S rRNA
tetracycline (30 mg) and vancomycin (30 mg). These anti-
gene sequence similarities of 95.1, 94.8, 94.6, 93.1, 92.4,
microbial discs were purchased from Oxoid.
90.1, 89.9 and 89.7 % to the type strains of Nocardioides
Biomass for molecular systematic studies and freeze-dried panacisoli GSoil 346T, Nocardioides aquiterrae GW-9T,
cells for chemotaxonomic studies were obtained after Aeromicrobium erythreum NRRL B-3381T, Marmoricola
growing cells in TSB at 28 uC for 7 days on a rotary shaker. aurantiacus BC 361T, Kribbella flavida DSM 17836T,
The analysis of peptidoglycan amino-acid composition and Actinopolymorpha singaporensis IM 7744T, Flindersiella
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88* Nocardioides albus KCTC 9186T (AF004988)
0.005 96 Nocardioides panzhihuensis KLBMP 1050T (HM153774)
100*
Nocardioides luteus KCTC 9575T (AF005007)
*
Nocardioides albertanoniae CD40127T (HE801966)
Nocardioides marinus CL-DD14T (DQ401093)
Nocardioides panacihumi Gsoil 616T (AB271053)
71 Nocardioides terrae VA15T (FJ423762)
Nocardioides insulae DS-51T (DQ786794)
80* Nocardioides alpinus Cr7-14T (GU784866)
Nocardioides furvisabuli SBS-26T (DQ411542)
72* Nocardioides exalbidus RC825T (AB273624)
99
Nocardioides hwasunensis HFW-21T (AM295258)
97* Nocardioides oleivorans DSM 16090T (AJ698724)
85* Nocardioides ganghwensis JC2055T (AY423718)
Nocardioides panacisoli GSoil 346T (FJ666101)
Nocardioides aestuarii JC2056T (AY423719)
Nocardioides tritolerans MSL-14T (EF466107)
99* Nocardioides maradonensis RP-B30T (FM998000)
87* Nocardioides ultimimeridianus RP-B26T (FM997998)
Nocardioides humi DCY24T (EF623863)
* Nocardioides simplex KCTC 9106T (AF005009)
78 57* Nocardioides ginsengisoli Gsoil 1124T (AB245396)
Nocardioides aromaticivorans H-1T (AB087721)
99*
Nocardioides caeni MN8T (FJ423551)
Nocardioides daeguensis 2C1-5T (HQ246164)
* Nocardioides nitrophenolicus NSP41T (AF005024)
72* Nocardioides kongjuensis A2-4T (DQ218275)
Nocardioides sediminis MSL-01T (EF466110)
99* Nocardioides terrigena DS-17T (EF363712)
62* Nocardioides aquiterrae GW-9T (AF529063)
82 Nocardioides pyridinolyticus OS4T (U61298)
51*
Nocardioides hankookensis DS-30T (EF555584)
Nocardioides hungaricus 1RaM5-12T (AM981198)
Nocardioides fonticola NAA-13T (EF626689)
Nocardioides plantarum NCIMB 12834T (AF005008)
90* Nocardioides ginsengagri BX5-10T (GQ339904)
74* Nocardioides marinquilinus CL-GY44T (JX164255)
Nocardioides aquaticus EL-17KT (X94145)
*
Nocardioides perillae I10A-01402T (JN869461)
Nocardioides bigeumensis MSL-19T (EF466114)
Nocardioides lentus KSL-17T (DQ121389)
Nocardioides islandensis MSL-26T (EF466123)
100* Nocardioides agariphilus MSL-28T (EF466113)
82* Nocardioides psychrotolerans RHLT2-1T (JF750425)
93* Nocardioides szechwanensis RHLT1-17T (JF750424)
Nocardioides kribbensis KSL-2T (AY835924)
Nocardioides lianchengensis D94-1T (HQ657322)
Nocardioides dokdonensis FR1436T (EF633986)
95* Nocardioides marinisabuli SBS-12T (AM422448)
86* Nocardioides salarius CL-Z59T (DQ401092)
82* Nocardioides basaltis J112T (EU143365)
Nocardioides caricicola YC6903T (FJ750845)
‘Nocardioides panaciterrulae’ Gsoil 958 (GQ339903)
69* Nocardioides ginsengisegetis Gsoil 485T (GQ339901)
78* Nocardioides koreensis MSL-09T (EF466115)
Nocardioides daphniae D287T (AM398438)
Nocardioides alkalitolerans KSL-1T (AY633969)
94* Nocardioides dubius KSL-104T (AY928902)
Nocardioides daejeonensis MJ31T (JF937066)
85* Nocardioides daedukensis MDN22T (FJ842646)
94* Nocardioides jensenii DSM 20641T (Z78210)
80* Nocardioides mesophilus MSL-22T (EF466117)
86* 86* Nocardioides iriomotensis IR27-S3T (AB544079)
Marmoricola bigeumensis MSL-05T (EF466120)
93* Marmoricola aurantiacus BC 361T (Y18629)
78
Marmoricola scoriae Sco-D01T (FN386750)
85* Marmoricola korecus Sco-A36T (FN386723)
93* Marmoricola aequoreus SST-45T (AM295338)
Nocardioides halotolerans MSL-23T (EF466122) Fig. 1. Neighbour-joining tree based on
100* Nocardioides dilutus MSL-11T (EF466121) an almost complete 16S rRNA sequence
Mumia flava MUSC 201T (KC907394) (1486 nt) showing the relationship between
‘Aeromicrobium massiliense’ JC14 (JF824798)
89* 100* Aeromicrobium tamlense SSW1-57T (DQ411541) strain MUSC201T and representatives of the
100* Aeromicrobium flavum TYLN1T (EF133690) family Nocardioidaceae. Bootstrap values
73 Aeromicrobium erythreum NRRL B-3381T (AF005021)
Aeromicrobium ponti HSW-1T (AM778683)
(.50 %) based on 1000 resampled datasets
96 70*
Aeromicrobium halocynthiae KME 001T (FJ042789) are shown at branch nodes. Bar, 5 substitu-
57 Aeromicrobium fastidiosum DSM 10552T (Z78209) tions per 1000 nucleotide positions. Asterisks
67 Aeromicrobium alkaliterrae KSL-107T (AY822044)
Aeromicrobium marinum DSM 15272T (ACLF01000927)
indicate that the corresponding nodes were
94* Aeromicrobium ginsengisoli Gsoil 098T (AB245394) also recovered using maximum-likelihood and
60* Aeromicrobium panaciterrae Gsoil 161T (AB245387) maximum-parsimony tree-making algorithms.
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endophytica EUM 378T and Thermasporomyces composti occurred at pH 5.0–10.0 (optimum pH 7.0–8.0), with 0–
I3T, respectively. Strain MUSC 201T formed a distinct clade 8 % NaCl (optimum 0–4 %) and at 20–36 uC (optimum
from the type strains of the genus Nocardioides at a low 28–32 uC). Hydrolysis of soluble starch, CM-cellulose and
nucleotide sequence similarity (91.9–95.1 %); this asso- chitin was positive, but hydrolysis of tributyrin (lipase),
ciation was supported by all of the different tree-making casein and xylan was negative. The morphological, cultural
algorithms used in this study. and physiological properties of strain MUSC 201T are given
in the genus and species descriptions. The organism can be
The total hydrolysate (4 M HCl, 100 uC, 16 h) of distinguished from members of the family Nocardioidaceae
peptidoglycan of strain MUSC 201T contained LL-diami- using different chemotaxonomic characteristics (Table 1).
nopimelic acid, glycine, glutamic acid and alanine in a
molar ratio of 1.5 : 0.9 : 1.0 : 1.5. The partial hydrolysate Strain MUSC 201T was similar to members of the genera of
(4 M HCl, 100 uC, 45 min) contained peptides L-Ala-D- the family Nocardioidaceae (Nocardioides, Aeromicrobium,
Glu, Gly-D-Ala, LL-Dpm-D-Ala and LL-Dpm-Gly (Schleifer Marmoricola, Kribbella, Actinopolymorpha, Flindersiella and
& Kandler, 1972; Schleifer, 1985). From these analytical Thermasporomyces), which contain LL-diaminopimelic acid
data, it was concluded that strain MUSC 201T contained as the diagnostic diamino acid. Based on the phylogenetic
the peptidoglycan type A3c, LL-Dpm-Gly. The cell-wall tree generated using the neighbour-joining algorithm, strain
sugars were galactose and rhamnose. The menaquinones MUSC 201T could also be assigned to the family Nocar-
detected were MK-9(H4) (89 %), MK-9 (1 %), MK-8(H4) dioidaceae as it formed a distinct clade with the type strains
(1 %), MK-9(H2) (1 %), MK-9(H6) (1 %) and MK-10(H4) of the genera Nocardioides, Aeromicrobium and Marmoricola
(traces). The polar lipids were diphosphatidylglycerol, (Fig. 1). The DNA G+C content of 72±0.1 mol% also fell
phosphatidylglycerol, phosphoglycolipid, glycolipid and within the range of DNA G+C contents within the family
four unknown phospholipids (Fig. S1, available in the Nocardioidaceae that range from 69.2 to 73 % (Table 1).
online Supplementary Material). The major cellular fatty Strain MUSC 201T was similar to members of genera such as
acids (.5 %) were C18 : 1v9c (30.8 %), C16 : 0 (24.1 %), 10- Aeromicrobium, Kribbella and Thermasporomyces in contain-
methyl C18 : 0 (13.9 %), C16 : 0 2-OH (7.6 %), C18 : 0 (5.5 %) ing the same predominant menaquinone MK-9(H4), but
and C17 : 0 (5.4 %) (Table S1). The G+C content of the DNA differed from members of genera such as Nocardioides and
was 72.0±0.1 mol%, as determined by HPLC analysis. Marmoricola in that it contained MK-8(H4) as a minor
Differential chemotaxonomic characteristics between strain menaquinone. Furthermore strain MUSC 201T could be
MUSC 201T and other genera belonging to the family differentiated from members of the genus Nocardioides by
Nocardioidaceae are summarized in Table 1. many phylogenetic, chemotaxonomic and phenotypic
properties (Table 1), e.g. strain MUSC 201T had low 16S
Cells were Gram-stain-positive, non-motile, aerobic, non- rRNA gene sequence similarities with members of the genus
spore-forming and irregular cocci or rod-shaped (Fig. 2). Nocardioides (91.9–95.1 %) and was separated from them by
Cells could occur singly, in pairs, in short chains or in small a long evolutionary distance in the phylogenetic tree (Fig. 1).
irregular clusters. Good growth was observed on ISP2 Furthermore strain MUSC 201T was significantly different
medium and nutrient agar after 7 days at 28 uC; cells grew from members of the genus Nocardioides and other genera of
moderately on SA, whereas cells grew poorly on AIA, SCA the family Nocardioidaceae in the fatty acid and polar lipid
and Luria–Bertani agar. Colonies were yellowish white on profiles, e.g. the polar lipid profile of strain MUSC 201T
most media tested. No aerial mycelia or diffusible pigments contained a phosphoglycolipid and a glycolipid that was not
were observed on any of the media. Cells were positive for detected in any of the other genera. For the fatty acids
catalase but negative for haemolytic activity. Growth profile, strain MUSC 201T was significantly different from
Table 1. Phenotypic and chemotaxonomic properties of strain MUSC201T and members of the family Nocardioidaceae
Strains: 1, Mumia flava gen. nov., sp. nov. MUSC 201T; 2, Nocardioides (data from Prauser, 1976; O’Donnell et al., 1982; Collins et al., 1989;
Tamura & Yokota, 1994; Park et al., 1999; Dastager et al., 2009; Cho et al., 2010); 3, Aeromicrobium (Miller et al., 1991; Tamura & Yokota, 1994;
Lee & Lee, 2008; Kim et al., 2010); 4, Marmoricola (Urzı̀ et al., 2000; Dastager et al., 2008; Lee & Lee, 2010; Lee et al., 2011). DPG,
diphosphatidylglycerol; GL, glycolipid; PG, phosphatidylglycerol; PGL, phosphoglycolipid; PI, phosphatidylinositol; PIM, phosphatidylinositol
mannoside; PC, phosphatidylcholine; PL, unknown phospholipid.
Characteristic 1 2 3 4
Major menaquinone MK-9(H4) MK-8(H4) MK-9(H4) MK-8(H4)
Major fatty acids C18 : 1v9c, C16 : 0, 10-methyl C18 : 0, iso-C16 : 0 C16 : 0, C16 : 0 2-OH, 10- C16 : 0, C18 : 1v9c,
methyl C18 : 0, C18 : 1v9c (iso-C16 : 0)*
Polar lipids DPG, PG, PL, PGL, GL PG PG, DPG PI, PG, DPG
*Major fatty acid of Marmoricola bigeumensis MSL-05T is C16 : 0.
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Cells are positive for catalase and negative for haemolytic
activities. Good growth is observed on ISP2 medium and
nutrient agar. Colonies are yellowish white on most media
tested. No aerial mycelia or diffusible pigments are
observed on any of the media. Grows at pH 5.0–10.0
(optimum pH 7.0–8.0), with 0–8 % NaCl (optimum 0–
4 %) and at 20–36 uC (optimum 28–32 uC). Hydrolysis of
soluble starch, CM-cellulose and chitin is positive; negative
for hydrolysis of tributyrin (lipase), casein and xylan. With
Biolog GEN III MicroPlates, the following compounds are
utilized as sole carbon sources: dextrin, maltose, trehalose,
cellobiose, gentiobiose, sucrose, turanose, melibiose, a-D-
glucose, D-mannose, D-fructose, D-galactose, 3-methyl
glucose, D-fucose, L-fucose, L-rhamnose, D-glucose 6-
phosphate, D-fructose 6-phosphate, D-galacturonic acid,
D-glucuronic acid, glucuronamide, L-lactic acid, citric acid,
Tween 40, a-hydroxybutyric acid, hydroxyl b-DL-butyric
Fig. 2. Scanning electron micrograph of cells from a 5-day-old acid, acetoacetic acid, propionic acid and acetic acid. The
culture of strain MUSC 201T grown at 28 6C on ISP 2. Bar, following compounds are not utilized as sole carbon
10 mm. sources: stachyose, raffinose, a-lactose, methyl b-D-gluc-
oside, D-salicin, N-acetyl-D-glucosamine, N-acetyl-b-D-
mannosamine, N-acetyl-D-galactosamine, N-acetyl-neura-
members of the genus Nocardioides, e.g. strain MUSC 201T minic acid, inosine, D-sorbitol, D-mannitol, D-arabitol,
contained iso-C18 : 1v9c (30.8 %), C16 : 0 (24.1 %), and 10- myo-inositol, glycerol, D-aspartic acid, D-serine, gelatin,
methyl C18 : 0 (13.9 %) as major fatty acids whereas N. glycyl L-proline, pectin, L-galactonic acid lactone, D-
panacisoli GSoil 346T contained iso-C16 : 0 (28.1 %), gluconic acid, mucic acid, quinic acid, D-saccharic acid,
C18 : 1v9c (12.8 %) and C17 : 1v8c (10.6 %) as major fatty p-hydroxylphenylacetic acid, methyl pyruvate, D-lactic acid
acids (Table S1). Furthermore strain MUSC 201T contained methyl ester, a-ketoglutaric acid, D-malic acid, L-malic
galactose and rhamnose as whole-cell sugars while species of acid, bromosuccinic acid, c-aminobutyric acid, a-ketobu-
the genus Nocardioides contained different whole-cell sugars tyric acid and formic acid. Sole nitrogen sources such as
such as ribose, glucose and galactose. Therefore, on the basis L-alanine, L-arginine, L-aspartic acid, L-glutamic acid, L-
of phylogenetic, chemotaxonomic and phenotypic profiles, histidine, L-pyroglutamic acid and L-serine are not utilized.
strain MUSC 201T is truly different from any existing genera In chemical sensitivity assays, cells are sensitive towards
in the family Nocardioidaceae and represents a novel species 1 % sodium lactate, troleandomycin, niaproof 4, vanco-
in a new genus of the family Nocardioidaceae, for which the mycin and sodium bromate, while cells are resistant to
name Mumia flava gen. nov., sp. nov. is proposed. fusidic acid, D-serine, rifamycin RV, minocycline, linco-
mycin, guanine hydrochloride, tetrazolium violet, tetra-
Description of Mumia gen. nov. zolium blue, nalidixic acid, lithium chloride, potassium
tellurite, aztreonam and sodium butyrate. Cells are resistant
Mumia (Mum9i.a. N.L. fem. n. Mumia derived from the to (per disc) erythromycin (15 mg), but sensitive to ampicillin
abbreviation MUM, for the Monash University Malaysia). (10 mg), ampicillin sulbactam (30 mg), cefotaxime (30 mg),
Aerobic, non-motile, non-spore-forming, Gram-stain-pos- cefuroxime (30 mg), cephalosporin (30 mg), chloramphenicol
itive actinobacteria of irregular coccoid to short-rod shape. (30 mg), ciprofloxacin (10 mg), gentamicin (20 mg), nalidixic
Cells occur singly, in pairs, in short chains or in small acid (30 mg), penicillin G (10 mg), streptomycin (10 mg),
irregular clusters. The predominant menaquinone is MK- tetracycline (30 mg) and vancomycin (30 mg).
9(H4). The polar lipids are diphosphatidylglycerol, phos- The type strain is MUSC 201T (5DSM 27763T5MCCC
phatidylglycerol, phosphoglycolipid, glycolipid and four 1A00646T5NBRC 109973T), which was isolated from
unknown phospholipids. The major cellular fatty acids are mangrove soil collected from Kuantan, the capital city of
C18 : 1v9c, C16 : 0 and 10-methyl C18 : 0. The peptidoglycan Pahang State in the Peninsular of Malaysia. The G+C
contains LL-diaminopimelic acid as diagnostic diamino acid content of the genomic DNA of strain MUSC 201T is
and the peptidoglycan type is A3c. The peptidoglycan cell 72±0.1 mol%.
wall contains LL-diaminopimelic, glycine, glutamic acid and
alanine. The cell-wall sugars are galactose and rhamnose.
Acknowledgements
Description of Mumia flava sp. nov. This work was supported by the University of Malaya for High
Impact Research Grant (UM-MOHE HIR Nature Microbiome Grant
Mumia flava (fla9va. L. fem. adj. flava yellow, referring to no. H-50001-A000027) to C. K.-G. and External Industry Grant
the colour of the colonies). (Biotek Abadi – Vote no. GBA-808138) awarded to L. L.-H. Authors
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On: Thu, 29 Oct 2015 23:41:23L.-H. Lee and others
are grateful to Dr Jean Euzéby and Professor Dr Bernhard Schink for Kuster, E. & Williams, S. T. (1964). Selection of media for isolation of
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