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International Journal of Cosmetic Science, 2004, 26, 149–156

Generation of volatile fatty acids by axillary bacteria1

A. G. James, D. Hyliands and H. Johnston2

Unilever R&D Colworth, Colworth House, Sharnbrook, Bedford MK44 1LQ, U.K.

Received 15 February 2004, Accepted 20 February 2004

Keywords: amino acid, axilla, malodour, Propionibacterium, Staphylococcus, volatile fatty acid

Synopsis Résumé
It is generally accepted that short-chain (C2–C5) Il est généralement accepté que les acides gras vola-
volatile fatty acids (VFAs) are among the causal tils à courtes chaı̂nes (C2–C5) (VFA’s) sont entre
molecules of axillary malodour. It is also widely autres les molécules à l’origine des mauvaises
acknowledged that malodour generation is attrib- odeurs axillaires. Il est aussi grandement admis que
utable to the biotransformation of odourless nat- la génération de ces odeurs est attribuable à la bio-
ural secretions, into volatile odorous products, by transformation de sécrétions naturelles sans odeurs
axillary bacteria. However, little information is en des produits odoriférants volatils par les bactéries
available on the biochemical origins of VFAs on axillaires. Cependant, les origines biochimiques des
axillary skin. In these studies, assay systems were VFAs sur les aisselles sont peu documentées. Dans
developed to investigate the generation of VFAs les études présentées ici, des systèmes de dosage ont
from substrates readily available to the bacteria été développés afin de rechercher la génération de
resident on axillary skin. Propionibacteria and sta- VFAs de substrats déjà disponibles aux bactéries rés-
phylococci were shown to ferment glycerol and identes des aisselles. Les Propionibacteria et Staphy-
lactic acid to the short-chain (C2–C3) VFAs, acetic locoques ont été montrés capables de fermenter le
and propionic acid. Furthermore, staphylococci are glycérol et l’acide lactique vers les VFA à courtes
capable of converting branched aliphatic amino chaı̂nes C2–C3, les acides acétique et propionique.
acids, such as leucine, to highly odorous short- De plus, les Staphylocoques son capables de conver-
chain (C4–C5) methyl-branched VFAs, such as tir les acides aminés aliphatiques tels que la leucine
isovaleric acid, which are traditionally associated en des courtes chaı̂nes méthylées (C4–C5) haute-
with the acidic note of axillary malodour. How- ment odoriférantes comme l’acide iso-valérique, tra-
ever, in vitro kinetic data indicates that these path- ditionnellement associées à la note acide des
ways contribute less to axillary VFA levels, than mauvaises odeurs axillaires. Cependant, les résultats
fatty acid biotransformations by a recently defined in vitro des cinétiques indiquent que ces transforma-
sub-group of the Corynebacterium genus, coryne- tions contribuent à des niveaux de VFAs moins
bacteria (A). The results of these studies provide élevés que celles des biotransformations des acides
new understanding on the biochemical origins of gras par un sous groupe du genre Corynebcaterium
VFA-based axillary malodour which, in turn, récemment défini, les Corynebacteria (A). Les résul-
should lead to the development of novel deodorant tats de ces études fournissent une nouvelle compré-
systems. hension des origines biochimiques des odeurs
axillaires à base de VFAs qui, à leur tour, devraient
conduire à de nouveaux systèmes déodorants.
Correspondence: A. G. James, Unilever R&D Colworth,
Bedford, U.K. Tel: +44 1234 222706; fax: +44 1234
222552; e-mail: Introduction
Presented in part at the 22nd IFSCC Congress, Edin-
burgh 2002. The generation of malodour on various sites of the
Present address: University of Abertay, Dundee, U.K. human body is caused by the microbial biotrans-

ª 2004 Society of Cosmetic Scientists and the Société Française de Cosmétologie 149
Generation of volatile fatty acids A. G. James et al.

formation of odourless natural secretions into vola- present metabolic problems to microorganisms, as
tile odorous molecules. On the skin surface, dis- the end-products of b-oxidation, isobutyric, isoval-
tinctive odours emanate, in particular, from the eric and 2-methylbutyric acids, may be non-utiliza-
underarm (axilla), where a large and permanent ble. Also present are fatty acids with internal
population of microorganisms thrives on secretions methyl branches on even-numbered carbon atoms.
from the eccrine, apocrine, apoeccrine and seba- These may also present metabolic problems, as the
ceous glands [1,2]. It is generally accepted that 2-methylacyl-CoA intermediates of b-oxidation may
short-medium chain (C2–C11) volatile fatty acids either be non-utilizable, or hinder chain-shortening
(VFAs) and 16-androstene steroids are among the such that metabolite leakage occurs [12]. In the
causal molecules of axillary malodour [3–5]. New axilla, long-chain fatty acids originate principally
information on the biochemical origins of 16- from the hydrolysis of the triglyceride component
androstenes on axillary skin, and the extent of of sebum, and potentially also apocrine lipid, by
their contribution to malodour, have recently been the action of bacterial lipases [13]. The biotrans-
reported [6]. The involvement of short-chain formation of triglycerides and fatty acids to VFAs
(C2–C5) VFAs in human body odours, including by axillary microorganisms, implicating a previ-
axillary malodour, has long been acknowledged; ously undefined sub-group of the Corynebacterium
latterly, medium chain (C6–C12) acids, in partic- genus, corynebacteria (A), is reported in a separate
ular the trans (E) isomer of 3-methyl-2-hexenoic article (A. G. James, J. Casey, D. Hyliands, and G.
acid (3M2H), have also been implicated [4]. While Mycock, World J. Microbiol. Biotechnol., in press).
little has been reported on the biochemical origins A further potential metabolic route to VFAs in
of VFAs on axillary skin, a notable exception is the axilla is the fermentation of glycerol, from
3M2H, which was shown by Spielman et al. [7] to triglyceride hydrolysis, and lactic acid, naturally
be carried to the skin surface non-covalently abundantly present on skin [14], to acetic (C2)
bound to two proteins, apocrine secretion odor- and propionic (C3) acids by resident microaerophi-
binding proteins 1 and 2 (ASOB1 and ASOB2). lic Propionibacterium species [15]. In propionibacte-
The ASOB2 protein was subsequently identified as rial fermentations, these substrates can be
apolipoprotein D (apoD), a member of the lipocalin converted to pyruvate, via glycolysis in the case
family of carrier proteins [8]. Recently, an appar- of glycerol, or by lactate dehydrogenase for
ently contradictory study by Natsch et al. [9] lactic acid. Pyruvate is then decarboxylated to
indicated that 3M2H, and the chemically related acetic acid, via acetyl-CoA, or converted to propi-
3-hydroxy-3-methylhexanoic acid, are instead onic acid, after carboxylation to oxaloacetate, and
covalently bound to free glutamine residues in subsequent decarboxylation to propionyl-CoA, via
fresh axillary secretions, and released by the action succinate, succinyl-CoA and methylmalonyl-CoA.
of corynebacterial N-acylglutamine aminoacylase. The proportion of C2 : C3 VFA produced is sub-
However, it was postulated that, once cleaved strate-dependent, with flux through each pathway
from their glutamine conjugates, the acids might regulated by the need to achieve overall redox
subsequently associate non-covalently with apoD. balance [15]. Acetic and propionic acids may also
Nevertheless, while the contribution of 3M2H and be produced, from glycerol and lactic acid, by fac-
3-hydroxy-3-methylhexanoic acid to axillary mal- ultatively anaerobic Staphylococcus species resident
odour is not disputed, there remains a lack of on axillary skin, probably involving alternative fer-
information on the ability of axillary bacteria to mentation pathways [15]. Several amino acids
generate VFAs via the biotransformation of readily likewise have the potential to be converted to
available cutaneous substrates. C2–C3 VFAs by propionibacteria and staphylococci,
A potential route to VFAs in the axilla is the via similar fermentation routes. However, certain
biotransformation of longer chain (C14–C30), struc- amino acids, specifically the branched aliphatic
turally unusual (odd carbon number and/or family (valine, leucine and isoleucine), may be
methyl-branched) fatty acids present in sebum metabolized, by alternative pathways, to the
[10,11], and possibly also apocrine sweat [A. G. highly odorous short-chain (C4–C5) methyl-
James, unpublished observation]. Sebaceous lipid branched VFAs, such as isovaleric (isoC5) acid
contains, in particular, large amounts of methyl- [16,17], traditionally associated with the acidic
branched fatty acids, the majority with the branch note of axillary malodour [3,4]. Although amino
in the iso- or anteiso-position. Potentially, these acids, including the branched aliphatic family, are

150 ª 2004 International Journal of Cosmetic Science, 26, 149–156

Generation of volatile fatty acids A. G. James et al.

present in eccrine sweat, they may also be provi- basal medium (g L)1) of KH2PO4 (1.6),
ded, on axillary skin, by the breakdown of protei- (NH4)2HPO4 (5.0), Na2SO4 (0.38), yeast nitrogen
naceous material from the keratinizing epidermis base (Difco, Le Pont de Claix, France) (3.35) and
and apocrine secretion. Many cutaneous bacteria, MgCl2Æ6H2O (0.5). For the fermentation assay, this
including micrococci, brevibacteria and, of partic- was supplemented with yeast extract (10 g L)1),
ular importance in the axilla, Staphylococcus epider- casamino acids (Difco) (2 g L)1), a-ketobutyric acid
midis and some propionibacteria, are known to (1 g L)1), Tween 80 (1 g L)1), anaerobic special
exhibit protease activity [13]. supplement (Oxoid) (2 vials L)1), guanidine
In this article, we describe in vitro studies on the (10 mg L)1), adenine (10 mg L)1), nicotinamide
formation of VFAs from substrates readily avail- (2 mg L)1), biotin (0.02 mg L)1), FeSO4 (1 mg L)1),
able to axillary bacteria, specifically glycerol, lactic CaCl2 (0.38 mg L)1), CoCl2 (0.24 mg L)1), MnSO4
acid and amino acids. We also report on a kinetic (0.38 mg L)1), ZnCl2 (0.34 mg L)1) and MnSO4
study designed to determine the relative contribu- (0.38 mg L)1). For the amino acid bioassay, the
tion of the main organisms and metabolic path- basal medium was supplemented (g L)1) with
ways to VFA-based axillary malodour. Tween 80 (0.1), glucose (5.0), CoCl2 (0.01), uracil
(0.02), alanine (0.1), arginine (0.1), aspartate
(0.1), cystine (0.1), glutamate (0.1), glycine (0.1),
lysine (0.1), phenylalanine (0.1), proline (0.1),
serine (0.1), threonine (0.1) and tyrosine (0.1).
General chemicals

Unless otherwise indicated, all general laboratory

Biotransformation assays
chemicals were obtained from Sigma-Aldrich Com-
pany (UK) Ltd. (Poole, Dorset, U.K.) The in vitro assay systems, for investigating VFA
generation by axillary bacteria, consisted of
250 mL baffled shake flasks, to which were added
Organisms and growth media
25–30 mL synthetic or semi-synthetic medium
A library of gram-positive bacteria (Corynebacteri- supplemented with glycerol (10.0 g L)1), l-lactic
um, Brevibacterium, Propionibacterium, Staphylococ- acid (10.0 g L)1 sodium salt), l-valine (1.0 g L)1),
cus and Micrococcus species), characteristic of the l-leucine (1.0 g L)1) or l-isoleucine (1.0 g L)1)
normal flora of axillary skin [1,18], was esta- substrate. Following inoculation with fresh bacter-
blished, including both culture collection strains ial biomass, to give starting optical densities (A590)
(National Collection of Type Cultures (NCTC); of 1.0–2.0, equivalent to log10 8–9 viable cells
National Collection of Industrial and Marine Bac- ml)1, assays were incubated aerobically or anaero-
teria Ltd. (NCIMB, Aberdeen, U.K.)) and in-house bically (9–13% CO2, <1% O2) at 35 C, with
axillary isolates. Bacterial isolates were classified, agitation (shaking incubator (Infors AG, Bottmin-
to genus level, by their ability to grow on selective gen, Germany) at 125 r.p.m.), and analyzed after
agar plates [18], and, where possible, to species 24–72 h. Fresh biomass was obtained by pre-grow-
level, by the use of ‘api STAPH’, ‘api CORYNE’ and ing bacteria aerobically (or anaerobically for the
‘rapid ID 32 A’ identification systems (bioMerieux, propionibacteria) at 35 C in 25–50 mL TSBT,
Marcy-1’Etoile, France). For long-term mainten- using 250 mL baffled shake flasks (agitated at
ance, all cultures were stored on ‘Protect’ cryo- 125 r.p.m. in shaking incubator (Infors AG)), then
genic beads (Technical Service Consultants Ltd., harvesting by centrifugation and resuspending in a
Heywood, Lancashire, U.K.) at )80 C; in the shor- minimal volume of synthetic or semi-synthetic
ter term, maintenance was on TSAT plates or TSBT medium. At the beginning and end of each assay
medium (30 g L)1 tryptone soya broth (Merck, incubation, culture purity and viability were deter-
Darmstadt, Germany); 10 g L)1 yeast extract (Ox- mined, either qualitatively, by streaking 1.0 lL
oid); 10 g L)1 Tween 80; ±20 g L)1 bacteriological culture medium onto TSAT plates, or quantita-
agar (Oxoid, Basingstoke, Hampshire, U.K.)) under tively, by total viable count analysis on TSAT plates,
aerobic or, for the propionibacteria, anaerobic con- following serial dilution in quarter-strength Ringers
ditions (9–13% CO2, <1% O2). For the biotransfor- solution. Plates were incubated aerobically (or
mation assays, a synthetic or semi-synthetic anaerobically, for the propionibacteria) at 35 C for
medium was employed, in each case based on a 48–72 h before being assessed. Upon completion of

ª 2004 International Journal of Cosmetic Science, 26, 149–156 151

Generation of volatile fatty acids A. G. James et al.

each assay incubation, VFA levels were deter- culture medium by centrifugation. A known
mined by capillary gas chromatography (GC). amount of internal standard (caproic acid, as the
sodium salt, from an aqueous stock solution) was
added to the decanted supernatant, which was
GC determination of VFAs
then acidified (pH 1–2) by the addition of 3 m HCl.
VFA levels in the biotransformation assays were VFAs were analyzed on a Perkin Elmer 8000 (Ser-
quantitatively determined by capillary GC analysis. ies 2) GC, essentially as described above.
Initially, 5 mL aliquots from each flask were trans-
ferred into universal tubes, a known amount of
Results and discussion
caproic acid (from a stock solution in ethyl acet-
ate) added as an internal standard, and the culture
Fermentation routes to VFAs
medium acidified (pH 1–2) by the addition of 3 m
HCl. Liquid–liquid extraction was then carried out In this study, representative axillary Propionibacteri-
using 10 mL ethyl acetate, with the organic and um (P. avidum NCTC 11864 and P. acnes isolate
aqueous phases being resolved by centrifugation. G62) and Staphylococcus species (S. epidermidis iso-
An aliquot of each organic (upper) phase was then lates W7 and DH1) were tested for their ability to
transferred to a sampling tube, prior to analysis on produce C2–C3 VFAs by fermentation of glycerol
a Perkin Elmer 8000 (Series 2) GC fitted with a and lactic acid, under anaerobic conditions; the
15 m · 0.32 mm (internal diameter) nitrotereph- results from the Propionibacterium incubations are
thalic acid modified PEG/siloxane copolymer presented in Table I. It is apparent that P. avidum
(FFAP) fused silica capillary column (film thick- NCTC 11864 generates significant quantities of
ness, 0.25 lm) (Quadrex Scientific, Nottingham, propionic (C3) acid from the fermentation of both
U.K.). The column was attached to the split-splitless tested substrates, with acetic (C2) acid also pro-
injector and flame ionization detector (FID) of the duced from lactic acid. In each case, the observed
GC; injector and detector temperatures were each C2 : C3 VFA yield was very close to the theoretical
300 C. Carrier gas for the column was helium ratio based on the requirement for redox balance
(0.4 kg cm)2), whereas hydrogen (1.2 kg cm)2) [15]. Glycerol was a poor substrate for P. acnes
and air (1.6 kg cm)2) supplied the FID. The tem- G62; however, in common with P. avidum NCTC
perature programme for VFA analysis was 80 C 11864, this organism generated significant levels
(2 min), 80–155 C (7.5 C min)1) and 155 C of acetic and propionic acid from lactic acid, with
(1 min). Sample size for injection was 1 lL. VFA yields again very close to the theoretical C2 : C3
levels were quantified by comparison of peak areas VFA ratio. In the Staphylococcus systems (data not
with known concentrations of both internal (cap- shown), it was observed that each of the tested
roic acid) and externally run standards (tailored S. epidermidis strains metabolized glycerol and lactic
mixtures of VFAs, of known concentration, in acid, forming large amounts of acetic and propionic
ethyl acetate). In some cases, notably for the fer- acid as fermentation products. In these cases, flux
mentation assay incubations, where acetic acid is probably through lactyl-CoA, which can be
levels required accurate determination, VFAs were dehydrated to acrylyl-CoA, and then reduced to
directly analyzed in the aqueous phase. Initially, propionyl-CoA, with reducing power provided by
bacterial cells were removed from 1 mL aliquots of the formation of acetic acid from a portion of the

Table I VFA generation by axillary Propionibacterium spp., as determined by GC

VFA products
Substrate C2 : C3 VFA Theoretical C2 : C3
Species (Code) (10 g L)1) C2 (g L)1) C3 (g L)1) (% mol : mol) VFA (% mol : mol)

P. avidum (NCTC 11864) Glycerol 0.1 11.3 1 : 99 0 : 100

Lactic acid (Na+) 1.4 3.5 33 : 67 33 : 67
P. acnes (G62) Glycerol 0.0 0.4 0 : 100 0 : 100
Lactic acid (Na+) 1.5 3.3 36 : 64 33 : 67

152 ª 2004 International Journal of Cosmetic Science, 26, 149–156

Generation of volatile fatty acids A. G. James et al.

Table II VFA generation by axillary

Staphylococcus spp., as determined by Isobutyric acid Isovaleric acid
GC (% mol : mol (% mol : mol
Code Species from valine) from leucine)

W15 S. sciuri 92.3 97.6

W18 S. simulans 34.0 74.8
G16 S. aureus 51.5 56.4
W7 S. epidermidis 25.0 44.7
NCTC 11041 S. cohnii 19.9 46.6
NCTC 11042 S. haemolyticus 16.1 44.8
NCTC 11320 S. hominis 10.8 36.2
W45 Unspeciated 10.9 32.0
W46 Unspeciated 10.2 26.5
W8 S. epidermidis 7.7 15.7
NCTC 07292 S. saprophyticus 5.9 17.5
NCTC 11044 S. warneri 2.5 18.9
W3 S. lugdunensis 4.9 12.2
W55 Unspeciated 0.8 14.3
W56 Unspeciated 4.3 6.3
NCTC 11043 S. xylosus 9.4 0.0

lactic acid utilized [15]. The results from this isovaleric (isoC5) acid, and isoleucine and 2-methyl-
study indicate that fermentation pathways by cuta- butyric (2-MeC4) acid. The data also clearly
neous propionibacteria and staphylococci generate indicated that VFA production by staphylococci
short-chain (C2–C3) VFAs on axillary skin. increases significantly with increasing oxygen avail-
Although these acids are not heavily implicated in ability. This implies that biotransformation activity
underarm odor, their presence probably contributes is directly linked to overall metabolic activity, as
to the pungency of the VFA cocktail in the axilla. growth was also optimal under aerobic conditions,
and is in contrast to some other microbiological sys-
tems, where methyl-branched VFAs are formed
Branched aliphatic amino acid biotransformations
anaerobically as end-products of amino acid
In these studies, representative axillary Staphylococ- fermentations [19]. The failure of Propionibacterium
cus (S. aureus isolate G16, S. epidermidis isolate W7 species to produce VFAs from branched aliphatic
and S. haemolyticus NCTC 11042), Corynebacterium amino acids was also unexpected, as isovaleric acid
(NCIMB 40928 and isolates G36, G38 & G40), production has been reported for both dairy [17]
Micrococcus (NCIMB 40927 and isolate DH3), Brevi- and skin-borne representatives [K. T. Holland, per-
bacterium (B. epidermidis NCTC 11083 and isolate sonal communication] of this genus. However, the
W11) and Propionibacterium species (P. avidum conversion of leucine to isovaleric acid has previ-
NCTC 11864 and P. acnes isolate G62) were ously been demonstrated by staphylococci in fer-
screened for their ability to generate dissimilatory mented sausage [20]. The results from the reported
metabolites from valine, leucine and isoleucine. studies provide evidence that human cutaneous sta-
Assays were set up aerobically or anaerobically, as phylococci are similarly capable of such biotransfor-
described above, or under microaerobic conditions mations, and thus contribute to VFA-based skin
(shaking incubator (Infors AG) at 35 r.p.m.), and odours, including axillary malodour.
incubated for up to 72 h. The results (data not A wider range of Staphylococcus strains (n ¼ 16)
shown) demonstrated that only Staphylococcus spe- was subsequently examined to confirm the univer-
cies can produce odorous metabolites from these sality of VFA generation, from branched aliphatic
amino acids, forming significant levels of methyl- amino acid biotransformations, by members of this
branched VFAs (C4–C5), probably by a combination genus. Each organism was incubated for 72 h in
of transaminase, and 2-oxo-acid dehydrogenase or the presence of valine or leucine, and the levels of
decarboxylase activities [16,17]. Clear precursor- VFA formation determined (Table II). It is apparent
product relationships were demonstrated between that net VFA production varies widely from cul-
valine and isobutyric (isoC4) acid, leucine and ture to culture, ranging from 0.0 to 97.6% molar

ª 2004 International Journal of Cosmetic Science, 26, 149–156 153

Generation of volatile fatty acids A. G. James et al.

organisms, namely staphylococci and corynebacte-

ria (A), and metabolic pathways, respectively
amino acid and long-chain fatty acid biotransfor-
mations, to methyl-branched VFAs in the axilla.
Rates of VFA generation by representative axillary
Staphylococcus (S. epidermidis isolate W7) and
Corynebacterium (NCIMB 40928 and isolate G40)
species were determined, from the branched ali-
phatic amino acids valine and leucine, or the
long-chain methyl-branched fatty acid, isopalmitic
(isoC16) acid. Assays were set up aerobically, as
described above, or by James et al. (World J. Micro-
biol. Biotechnol., in press), except that samples
Figure 1 Branched aliphatic amino acid metabolism by
axillary bacteria.
were removed for VFA determination after 2, 5, 8,
12 and 24 h, and assay volumes were increased
conversion. Subsequent experimentation, using to 50 mL to compensate for this. Rates of product
isobutyric (isoC4), isovaleric (isoC5) and 2-methyl- formation were calculated as lg total VFA gener-
butyric (2-MeC4) acid as substrates, showed that ated per 109 viable cells (colony forming units) per
this was due to differences in the extent of VFA hour (lg 10)9 CFU h)1), based on culture viability
generation per se, rather than any further utiliza- at the beginning of each assay incubation. The
tion (data not shown). These experiments also results of this in vitro study indicate that rates of
showed that corynebacteria are incapable of meta- VFA generation, per unit cell number, by staphylo-
bolizing valine, leucine and isoleucine, or isobutyr- cocci, are significantly lower (7–10-fold) than by
ic (isoC4), isovaleric (isoC5) and 2-methylbutyric fatty acid-catabolizing corynebacteria (Table III).
(2-MeC4) acid, whereas most micrococci and brevi- Therefore, while it is likely that the metabolism of
bacteria can fully catabolize these amino acids and branched aliphatic amino acids by staphylococci
the corresponding methyl-branched VFAs (A. G. contributes to VFA-based axillary malodour, this
James et al., World J. Microbiol. Biotechnol., in probably does not represent the major biotransfor-
press). These results, summarized pictorially in mation route to these odorants.
Fig. 1, confirmed that VFA formation in the axilla
is a dynamic process, with some cutaneous micro-
Concluding comments
organisms capable of fully catabolizing these odor-
ants and their precursors. However, the The results from these in vitro studies provide evi-
prevalence and density of micrococci and brevibac- dence that human cutaneous staphylococci gener-
teria in the axilla are probably too low for them to ate malodorous short-chain (C4–C5) methyl-
have a significant effect on net VFA levels [18]. branched VFAs, and thus contribute to VFA-based
skin odours, including axillary malodour. The high
rates of carriage and prevalence of these bacteria on
Quantitation of VFA generation by axillary
normal human skin [21], including the axilla
[1,18], support this view. However, in vivo studies
A limited in vitro kinetic study was designed to indicate no direct association between staphylococ-
investigate the relative contribution of the main cal numbers and malodour intensity in the axilla, in

Table III Kinetics of VFA genera-

Rate tion by axillary Corynebacterium and
Substrate VFA (lg 10)9 Staphylococcus spp., as determined
Genus Species (Code) (1.0 g L)1) products CFU h)1) by GC

Staphylococcus S. epidermidis (W7) Leucine isoC5 7.0

Valine isoC4 4.4
Corynebacterium Unspeciated (G40) Isopalmitic acid isoC4, isoC6 48.8
Unspeciated (NCIMB 40928) Isopalmitic acid isoC4, isoC6 49.1

154 ª 2004 International Journal of Cosmetic Science, 26, 149–156

Generation of volatile fatty acids A. G. James et al.

Figure 2 Formation and utilization of VFAs by axillary bacteria.

contrast to corynebacteria, where such a relation- Microbiol. Biotechnol., in press), provide new
ship exists [18]. An explanation for this may be the understanding on the biochemical origins of VFA-
highly variable levels of VFA generation observed based axillary malodour (Fig. 2) which, in turn,
for different Staphylococcus strains (Table II). Even should lead to the development of novel deodorant
within the single species S. epidermidis, the most pre- systems. Furthermore, consideration of these arti-
valent on human skin [21], variation among iso- cles alongside those published by Austin and Ellis
lates (W7 and W8) was apparent. Further to this, [6], on 16-androstene steroids, and Natsch et al.
the reported in vitro kinetic data indicates that these [9], on 3M2H and 3-hydroxy-3-methylhexanoic
pathways contribute less to axillary VFA levels, acid, implies a recent step-change in our under-
than fatty acid biotransformations by a recently standing of axillary malodour.
defined sub-group of the Corynebacterium genus, co-
rynebacteria (A). However, branched aliphatic
amino acid biotransformations may be an import-
ant route to VFA-based malodour in the axillae of The authors would like to thank Prof. Keith Hol-
Staphylococcus-dominated high odor males, in land (University of Leeds, Leeds, U.K.) for reading
female axillae where carriage and prevalence of co- and commenting on aspects of this manuscript
rynebacteria are lower [D. Taylor, personal commu- and providing valued advice on scientific content.
nication], and on feet, where proteins are believed
to be the odor precursor molecules [22], and isoval-
eric acid the primary odorant [23].
The combined results of the reported studies, 1. Leyden, J.J., McGinley, K.J., Holzle, E. et al. The
and related findings by James et al. (World J. microbiology of the human axilla and its relationship

ª 2004 International Journal of Cosmetic Science, 26, 149–156 155

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to axillary odor. J. Invest. Dermatol. 77, 413–416 13. Holland, K.T. Nutrition of cutaneous resident micro-
(1981). organisms. In: The Skin Microflora and Microbial
2. Parekh, J.C. Axillary odor: its physiology, microbio- Skin Disease (Noble, W.C., ed.), pp. 33–72. Cam-
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