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
It is generally accepted that short-chain (C2–C5)
volatile fatty acids (VFAs) are among the causal
molecules of axillary malodour. It is also widely
acknowledged that malodour generation is attrib-
utable to the biotransformation of odourless nat-
ural secretions, into volatile odorous products, by
axillary bacteria. However, little information is
available on the biochemical origins of VFAs on
axillary skin. In these studies, assay systems were
developed to investigate the generation of VFAs
from substrates readily available to the bacteria
resident on axillary skin. Propionibacteria and sta-
phylococci were shown to ferment glycerol and
lactic acid to the short-chain (C2–C3) VFAs, acetic
and propionic acid. Furthermore, staphylococci are
capable of converting branched aliphatic amino
acids, such as leucine, to highly odorous short-
chain (C4–C5) methyl-branched VFAs, such as
isovaleric acid, which are traditionally associated
with the acidic note of axillary malodour. How-
ever, in vitro kinetic data indicates that these path-
ways contribute less to axillary VFA levels, than
fatty acid biotransformations by a recently defined
sub-group of the Corynebacterium genus, coryne-
bacteria (A). The results of these studies provide
new understanding on the biochemical origins of
VFA-based axillary malodour which, in turn,
should lead to the development of novel deodorant
systems.
Correspondence: A. G. James, Unilever R&D Colworth,
Bedford, U.K. Tel: +44 1234 222706; fax: +44 1234
222552; e-mail: gordon.james@unilever.com
1
Presented in part at the 22nd IFSCC Congress, Edin-
burgh 2002.
2
Present address: University of Abertay, Dundee, U.K.
ª 2004 Society of Cosmetic Scientists and the Société Française de Cosmétologie
Résumé
Il est généralement accepté que les acides gras vola-
tils à courtes chaı̂nes (C2–C5) (VFA’s) sont entre
autres les molécules à l’origine des mauvaises
odeurs axillaires. Il est aussi grandement admis que
la génération de ces odeurs est attribuable à la bio-
transformation de sécrétions naturelles sans odeurs
en des produits odoriférants volatils par les bactéries
axillaires. Cependant, les origines biochimiques des
VFAs sur les aisselles sont peu documentées. Dans
les études présentées ici, des systèmes de dosage ont
été développés afin de rechercher la génération de
VFAs de substrats déjà disponibles aux bactéries rés-
identes des aisselles. Les Propionibacteria et Staphy-
locoques ont été montrés capables de fermenter le
glycérol et l’acide lactique vers les VFA à courtes
chaı̂nes C2–C3, les acides acétique et propionique.
De plus, les Staphylocoques son capables de conver-
tir les acides aminés aliphatiques tels que la leucine
en des courtes chaı̂nes méthylées (C4–C5) haute-
ment odoriférantes comme l’acide iso-valérique, tra-
ditionnellement associées à la note acide des
mauvaises odeurs axillaires. Cependant, les résultats
in vitro des cinétiques indiquent que ces transforma-
tions contribuent à des niveaux de VFAs moins
élevés que celles des biotransformations des acides
gras par un sous groupe du genre Corynebcaterium
récemment défini, les Corynebacteria (A). Les résul-
tats de ces études fournissent une nouvelle compré-
hension des origines biochimiques des odeurs
axillaires à base de VFAs qui, à leur tour, devraient
conduire à de nouveaux systèmes déodorants.
Introduction
The generation of malodour on various sites of the
human body is caused by the microbial biotrans-
149
Generation of volatile fatty acids
formation of odourless natural secretions into vola-
tile odorous molecules. On the skin surface, dis-
tinctive odours emanate, in particular, from the
underarm (axilla), where a large and permanent
population of microorganisms thrives on secretions
from the eccrine, apocrine, apoeccrine and seba-
ceous glands [1,2]. It is generally accepted that
short-medium chain (C2–C11) volatile fatty acids
(VFAs) and 16-androstene steroids are among the
causal molecules of axillary malodour [3–5]. New
information on the biochemical origins of 16-
androstenes on axillary skin, and the extent of
their contribution to malodour, have recently been
reported [6]. The involvement of short-chain
(C2–C5) VFAs in human body odours, including
axillary malodour, has long been acknowledged;
latterly, medium chain (C6–C12) acids, in partic-
ular the trans (E) isomer of 3-methyl-2-hexenoic
acid (3M2H), have also been implicated [4]. While
little has been reported on the biochemical origins
of VFAs on axillary skin, a notable exception is
3M2H, which was shown by Spielman et al. [7] to
be carried to the skin surface non-covalently
bound to two proteins, apocrine secretion odor-
binding proteins 1 and 2 (ASOB1 and ASOB2).
The ASOB2 protein was subsequently identified as
apolipoprotein D (apoD), a member of the lipocalin
family of carrier proteins [8]. Recently, an appar-
ently contradictory study by Natsch et al. [9]
indicated that 3M2H, and the chemically related
3-hydroxy-3-methylhexanoic acid, are instead
covalently bound to free glutamine residues in
fresh axillary secretions, and released by the action
of corynebacterial N-acylglutamine aminoacylase.
However, it was postulated that, once cleaved
from their glutamine conjugates, the acids might
subsequently associate non-covalently with apoD.
Nevertheless, while the contribution of 3M2H and
3-hydroxy-3-methylhexanoic acid to axillary mal-
odour is not disputed, there remains a lack of
information on the ability of axillary bacteria to
generate VFAs via the biotransformation of readily
available cutaneous substrates.
A potential route to VFAs in the axilla is the
biotransformation of longer chain (C14–C30), struc-
turally unusual (odd carbon number and/or
methyl-branched) fatty acids present in sebum
[10,11], and possibly also apocrine sweat [A. G.
James, unpublished observation]. Sebaceous lipid
contains, in particular, large amounts of methyl-
branched fatty acids, the majority with the branch
in the iso- or anteiso-position. Potentially, these
150 ª 2004 International Journal of Cosmetic Science, 26, 149–156
A. G. James et al.
present metabolic problems to microorganisms, as
the end-products of b-oxidation, isobutyric, isoval-
eric and 2-methylbutyric acids, may be non-utiliza-
ble. Also present are fatty acids with internal
methyl branches on even-numbered carbon atoms.
These may also present metabolic problems, as the
2-methylacyl-CoA intermediates of b-oxidation may
either be non-utilizable, or hinder chain-shortening
such that metabolite leakage occurs [12]. In the
axilla, long-chain fatty acids originate principally
from the hydrolysis of the triglyceride component
of sebum, and potentially also apocrine lipid, by
the action of bacterial lipases [13]. The biotrans-
formation of triglycerides and fatty acids to VFAs
by axillary microorganisms, implicating a previ-
ously undefined sub-group of the Corynebacterium
genus, corynebacteria (A), is reported in a separate
article (A. G. James, J. Casey, D. Hyliands, and G.
Mycock, World J. Microbiol. Biotechnol., in press).
A further potential metabolic route to VFAs in
the axilla is the fermentation of glycerol, from
triglyceride hydrolysis, and lactic acid, naturally
abundantly present on skin [14], to acetic (C2)
and propionic (C3) acids by resident microaerophi-
lic Propionibacterium species [15]. In propionibacte-
rial fermentations, these substrates can be
converted to pyruvate, via glycolysis in the case
of glycerol, or by lactate dehydrogenase for
lactic acid. Pyruvate is then decarboxylated to
acetic acid, via acetyl-CoA, or converted to propi-
onic acid, after carboxylation to oxaloacetate, and
subsequent decarboxylation to propionyl-CoA, via
succinate, succinyl-CoA and methylmalonyl-CoA.
The proportion of C2 : C3 VFA produced is sub-
strate-dependent, with flux through each pathway
regulated by the need to achieve overall redox
balance [15]. Acetic and propionic acids may also
be produced, from glycerol and lactic acid, by fac-
ultatively anaerobic Staphylococcus species resident
on axillary skin, probably involving alternative fer-
mentation pathways [15]. Several amino acids
likewise have the potential to be converted to
C2–C3 VFAs by propionibacteria and staphylococci,
via similar fermentation routes. However, certain
amino acids, specifically the branched aliphatic
family (valine, leucine and isoleucine), may be
metabolized, by alternative pathways, to the
highly odorous short-chain (C4–C5) methyl-
branched VFAs, such as isovaleric (isoC5) acid
[16,17], traditionally associated with the acidic
note of axillary malodour [3,4]. Although amino
acids, including the branched aliphatic family, are
Generation of volatile fatty acids
present in eccrine sweat, they may also be provi-
ded, on axillary skin, by the breakdown of protei-
naceous material from the keratinizing epidermis
and apocrine secretion. Many cutaneous bacteria,
including micrococci, brevibacteria and, of partic-
ular importance in the axilla, Staphylococcus epider-
midis and some propionibacteria, are known to
exhibit protease activity [13].
In this article, we describe in vitro studies on the
formation of VFAs from substrates readily avail-
able to axillary bacteria, specifically glycerol, lactic
acid and amino acids. We also report on a kinetic
study designed to determine the relative contribu-
tion of the main organisms and metabolic path-
ways to VFA-based axillary malodour.
Experimental
General chemicals
Unless otherwise indicated, all general laboratory
chemicals were obtained from Sigma-Aldrich Com-
pany (UK) Ltd. (Poole, Dorset, U.K.)
Organisms and growth media
A library of gram-positive bacteria (Corynebacteri-
um, Brevibacterium, Propionibacterium, Staphylococ-
cus and Micrococcus species), characteristic of the
normal flora of axillary skin [1,18], was esta-
blished, including both culture collection strains
(National Collection of Type Cultures (NCTC);
National Collection of Industrial and Marine Bac-
teria Ltd. (NCIMB, Aberdeen, U.K.)) and in-house
axillary isolates. Bacterial isolates were classified,
to genus level, by their ability to grow on selective
agar plates [18], and, where possible, to species
level, by the use of ‘api STAPH’, ‘api CORYNE’ and
‘rapid ID 32 A’ identification systems (bioMerieux,
Marcy-1’Etoile, France). For long-term mainten-
ance, all cultures were stored on ‘Protect’ cryo-
genic beads (Technical Service Consultants Ltd.,
Heywood, Lancashire, U.K.) at )80 C; in the shor-
ter term, maintenance was on TSAT plates or TSBT
medium (30 g L)1 tryptone soya broth (Merck,
Darmstadt, Germany); 10 g L)1 yeast extract (Ox-
oid); 10 g L)1 Tween 80; ±20 g L)1 bacteriological
agar (Oxoid, Basingstoke, Hampshire, U.K.)) under
aerobic or, for the propionibacteria, anaerobic con-
ditions (9–13% CO2, <1% O2). For the biotransfor-
mation assays, a synthetic or semi-synthetic
medium was employed, in each case based on a
ª 2004 International Journal of Cosmetic Science, 26, 149–156
A. G. James et al.
basal medium (g L)1) of KH2PO4 (1.6),
(NH4)2HPO4 (5.0), Na2SO4 (0.38), yeast nitrogen
base (Difco, Le Pont de Claix, France) (3.35) and
MgCl2Æ6H2O (0.5). For the fermentation assay, this
was supplemented with yeast extract (10 g L)1),
casamino acids (Difco) (2 g L)1), a-ketobutyric acid
(1 g L)1), Tween 80 (1 g L)1), anaerobic special
supplement (Oxoid) (2 vials L)1), guanidine
(10 mg L)1), adenine (10 mg L)1), nicotinamide
(2 mg L)1), biotin (0.02 mg L)1), FeSO4 (1 mg L)1),
CaCl2 (0.38 mg L)1), CoCl2 (0.24 mg L)1), MnSO4
(0.38 mg L)1), ZnCl2 (0.34 mg L)1) and MnSO4
(0.38 mg L)1). For the amino acid bioassay, the
basal medium was supplemented (g L)1) with
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).
Biotransformation assays
The in vitro assay systems, for investigating VFA
generation by axillary bacteria, consisted of
250 mL baffled shake flasks, to which were added
25–30 mL synthetic or semi-synthetic medium
supplemented with glycerol (10.0 g L)1), l-lactic
acid (10.0 g L)1 sodium salt), l-valine (1.0 g L)1),
l-leucine (1.0 g L)1) or l-isoleucine (1.0 g L)1)
substrate. Following inoculation with fresh bacter-
ial biomass, to give starting optical densities (A590)
of 1.0–2.0, equivalent to
log10 8–9 viable cells
ml)1, assays were incubated aerobically or anaero-
bically (9–13% CO2, <1% O2) at 35 C, with
agitation (shaking incubator (Infors AG, Bottmin-
gen, Germany) at 125 r.p.m.), and analyzed after
24–72 h. Fresh biomass was obtained by pre-grow-
ing bacteria aerobically (or anaerobically for the
propionibacteria) at 35 C in 25–50 mL TSBT,
using 250 mL baffled shake flasks (agitated at
125 r.p.m. in shaking incubator (Infors AG)), then
harvesting by centrifugation and resuspending in a
minimal volume of synthetic or semi-synthetic
medium. At the beginning and end of each assay
incubation, culture purity and viability were deter-
mined, either qualitatively, by streaking 1.0 lL
culture medium onto TSAT plates, or quantita-
tively, by total viable count analysis on TSAT plates,
following serial dilution in quarter-strength Ringers
solution. Plates were incubated aerobically (or
anaerobically, for the propionibacteria) at 35 C for
48–72 h before being assessed. Upon completion of
151
Generation of volatile fatty acids
each assay incubation, VFA levels were deter-
mined by capillary gas chromatography (GC).
GC determination of VFAs
VFA levels in the biotransformation assays were
quantitatively determined by capillary GC analysis.
Initially, 5 mL aliquots from each flask were trans-
ferred into universal tubes, a known amount of
caproic acid (from a stock solution in ethyl acet-
ate) added as an internal standard, and the culture
medium acidified (pH 1–2) by the addition of 3 m
HCl. Liquid–liquid extraction was then carried out
using 10 mL ethyl acetate, with the organic and
aqueous phases being resolved by centrifugation.
An aliquot of each organic (upper) phase was then
transferred to a sampling tube, prior to analysis on
a Perkin Elmer 8000 (Series 2) GC fitted with a
15 m · 0.32 mm (internal diameter) nitrotereph-
thalic acid modified PEG/siloxane copolymer
(FFAP) fused silica capillary column (film thick-
ness, 0.25 lm) (Quadrex Scientific, Nottingham,
U.K.). The column was attached to the split-splitless
injector and flame ionization detector (FID) of the
GC; injector and detector temperatures were each
300 C. Carrier gas for the column was helium
(0.4 kg cm)2), whereas hydrogen (1.2 kg cm)2)
and air (1.6 kg cm)2) supplied the FID. The tem-
perature programme for VFA analysis was 80 C
(2 min), 80–155 C (7.5 C min)1) and 155 C
(1 min). Sample size for injection was 1 lL. VFA
levels were quantified by comparison of peak areas
with known concentrations of both internal (cap-
roic acid) and externally run standards (tailored
mixtures of VFAs, of known concentration, in
ethyl acetate). In some cases, notably for the fer-
mentation assay incubations, where acetic acid
levels required accurate determination, VFAs were
directly analyzed in the aqueous phase. Initially,
bacterial cells were removed from 1 mL aliquots of
Table I VFA generation by axillary Propionibacterium spp., as determined by GC
VFA products
Substrate
Species (Code) (10 g L)1) C2 (g L)1)
P. avidum (NCTC 11864) Glycerol 0.1
Lactic acid (Na+) 1.4
P. acnes (G62) Glycerol 0.0
Lactic acid (Na+) 1.5
152
A. G. James et al.
culture medium by centrifugation. A known
amount of internal standard (caproic acid, as the
sodium salt, from an aqueous stock solution) was
added to the decanted supernatant, which was
then acidified (pH 1–2) by the addition of 3 m HCl.
VFAs were analyzed on a Perkin Elmer 8000 (Ser-
ies 2) GC, essentially as described above.
Results and discussion
Fermentation routes to VFAs
In this study, representative axillary Propionibacteri-
um (P. avidum NCTC 11864 and P. acnes isolate
G62) and Staphylococcus species (S. epidermidis iso-
lates W7 and DH1) were tested for their ability to
produce C2–C3 VFAs by fermentation of glycerol
and lactic acid, under anaerobic conditions; the
results from the Propionibacterium incubations are
presented in Table I. It is apparent that P. avidum
NCTC 11864 generates significant quantities of
propionic (C3) acid from the fermentation of both
tested substrates, with acetic (C2) acid also pro-
duced from lactic acid. In each case, the observed
C2 : C3 VFA yield was very close to the theoretical
ratio based on the requirement for redox balance
[15]. Glycerol was a poor substrate for P. acnes
G62; however, in common with P. avidum NCTC
11864, this organism generated significant levels
of acetic and propionic acid from lactic acid, with
yields again very close to the theoretical C2 : C3
VFA ratio. In the Staphylococcus systems (data not
shown), it was observed that each of the tested
S. epidermidis strains metabolized glycerol and lactic
acid, forming large amounts of acetic and propionic
acid as fermentation products. In these cases, flux
is probably through lactyl-CoA, which can be
dehydrated to acrylyl-CoA, and then reduced to
propionyl-CoA, with reducing power provided by
the formation of acetic acid from a portion of the
Observed
C2 : C3 VFA Theoretical C2 : C3
C3 (g L)1) (% mol : mol) VFA (% mol : mol)
11.3 1 : 99 0 : 100
3.5 33 : 67 33 : 67
0.4 0 : 100 0 : 100
3.3 36 : 64 33 : 67
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
study indicate that fermentation pathways by cuta-
neous propionibacteria and staphylococci generate
short-chain (C2–C3) VFAs on axillary skin.
Although these acids are not heavily implicated in
underarm odor, their presence probably contributes
to the pungency of the VFA cocktail in the axilla.
Branched aliphatic amino acid biotransformations
In these studies, representative axillary Staphylococ-
cus (S. aureus isolate G16, S. epidermidis isolate W7
and S. haemolyticus NCTC 11042), Corynebacterium
(NCIMB 40928 and isolates G36, G38 & G40),
Micrococcus (NCIMB 40927 and isolate DH3), Brevi-
bacterium (B. epidermidis NCTC 11083 and isolate
W11) and Propionibacterium species (P. avidum
NCTC 11864 and P. acnes isolate G62) were
screened for their ability to generate dissimilatory
metabolites from valine, leucine and isoleucine.
Assays were set up aerobically or anaerobically, as
described above, or under microaerobic conditions
(shaking incubator (Infors AG) at 35 r.p.m.), and
incubated for up to 72 h. The results (data not
shown) demonstrated that only Staphylococcus spe-
cies can produce odorous metabolites from these
amino acids, forming significant levels of methyl-
branched VFAs (C4–C5), probably by a combination
of transaminase, and 2-oxo-acid dehydrogenase or
decarboxylase activities [16,17]. Clear precursor-
product relationships were demonstrated between
valine and isobutyric (isoC4) acid, leucine and
isovaleric (isoC5) acid, and isoleucine and 2-methyl-
butyric (2-MeC4) acid. The data also clearly
indicated that VFA production by staphylococci
increases significantly with increasing oxygen avail-
ability. This implies that biotransformation activity
is directly linked to overall metabolic activity, as
growth was also optimal under aerobic conditions,
and is in contrast to some other microbiological sys-
tems, where methyl-branched VFAs are formed
anaerobically as end-products of amino acid
fermentations [19]. The failure of Propionibacterium
species to produce VFAs from branched aliphatic
amino acids was also unexpected, as isovaleric acid
production has been reported for both dairy [17]
and skin-borne representatives [K. T. Holland, per-
sonal communication] of this genus. However, the
conversion of leucine to isovaleric acid has previ-
ously been demonstrated by staphylococci in fer-
mented sausage [20]. The results from the reported
studies provide evidence that human cutaneous sta-
phylococci are similarly capable of such biotransfor-
mations, and thus contribute to VFA-based skin
odours, including axillary malodour.
A wider range of Staphylococcus strains (n ¼ 16)
was subsequently examined to confirm the univer-
sality of VFA generation, from branched aliphatic
amino acid biotransformations, by members of this
genus. Each organism was incubated for 72 h in
the presence of valine or leucine, and the levels of
VFA formation determined (Table II). It is apparent
that net VFA production varies widely from cul-
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
Figure 1 Branched aliphatic amino acid metabolism by
axillary bacteria.
conversion. Subsequent experimentation, using
isobutyric (isoC4), isovaleric (isoC5) and 2-methyl-
butyric (2-MeC4) acid as substrates, showed that
this was due to differences in the extent of VFA
generation per se, rather than any further utiliza-
tion (data not shown). These experiments also
showed that corynebacteria are incapable of meta-
bolizing valine, leucine and isoleucine, or isobutyr-
ic (isoC4), isovaleric (isoC5) and 2-methylbutyric
(2-MeC4) acid, whereas most micrococci and brevi-
bacteria can fully catabolize these amino acids and
the corresponding methyl-branched VFAs (A. G.
James et al., World J. Microbiol. Biotechnol., in
press). These results, summarized pictorially in
Fig. 1, confirmed that VFA formation in the axilla
is a dynamic process, with some cutaneous micro-
organisms capable of fully catabolizing these odor-
ants and their precursors. However, the
prevalence and density of micrococci and brevibac-
teria in the axilla are probably too low for them to
have a significant effect on net VFA levels [18].
Quantitation of VFA generation by axillary
bacteria
A limited in vitro kinetic study was designed to
investigate the relative contribution of the main
Substrate
Genus Species (Code) (1.0 g L)1)
Staphylococcus S. epidermidis (W7) Leucine
Valine
Corynebacterium Unspeciated (G40) Isopalmitic acid isoC4, isoC6 48.8
Unspeciated (NCIMB 40928) Isopalmitic acid isoC4, isoC6 49.1
154
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
were removed for VFA determination after 2, 5, 8,
12 and 24 h, and assay volumes were increased
to 50 mL to compensate for this. Rates of product
formation were calculated as lg total VFA gener-
ated per 109 viable cells (colony forming units) per
hour (lg 10)9 CFU h)1), based on culture viability
at the beginning of each assay incubation. The
results of this in vitro study indicate that rates of
VFA generation, per unit cell number, by staphylo-
cocci, are significantly lower (
7–10-fold) than by
fatty acid-catabolizing corynebacteria (Table III).
Therefore, while it is likely that the metabolism of
branched aliphatic amino acids by staphylococci
contributes to VFA-based axillary malodour, this
probably does not represent the major biotransfor-
mation route to these odorants.
Concluding comments
The results from these in vitro studies provide evi-
dence that human cutaneous staphylococci gener-
ate malodorous short-chain (C4–C5) methyl-
branched VFAs, and thus contribute to VFA-based
skin odours, including axillary malodour. The high
rates of carriage and prevalence of these bacteria on
normal human skin [21], including the axilla
[1,18], support this view. However, in vivo studies
indicate no direct association between staphylococ-
cal numbers and malodour intensity in the axilla, in
Table III Kinetics of VFA genera-
Rate tion by axillary Corynebacterium and
VFA (lg 10)9 Staphylococcus spp., as determined
products CFU h)1) by GC
isoC5 7.0
isoC4 4.4
ª 2004 International Journal of Cosmetic Science, 26, 149–156
Generation of volatile fatty acids
Figure 2 Formation and utilization of VFAs by axillary bacteria.
contrast to corynebacteria, where such a relation-
ship exists [18]. An explanation for this may be the
highly variable levels of VFA generation observed
for different Staphylococcus strains (Table II). Even
within the single species S. epidermidis, the most pre-
valent on human skin [21], variation among iso-
lates (W7 and W8) was apparent. Further to this,
the reported in vitro kinetic data indicates that these
pathways contribute less to axillary VFA levels,
than fatty acid biotransformations by a recently
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
Staphylococcus-dominated high odor males, in
female axillae where carriage and prevalence of co-
rynebacteria are lower [D. Taylor, personal commu-
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,
and related findings by James et al. (World J.
ª 2004 International Journal of Cosmetic Science, 26, 149–156
A. G. James et al.
Microbiol. Biotechnol., in press), provide new
understanding on the biochemical origins of VFA-
based axillary malodour (Fig. 2) which, in turn,
should lead to the development of novel deodorant
systems. Furthermore, consideration of these arti-
cles alongside those published by Austin and Ellis
[6], on 16-androstene steroids, and Natsch et al.
[9], on 3M2H and 3-hydroxy-3-methylhexanoic
acid, implies a recent step-change in our under-
standing of axillary malodour.
Acknowledgements
The authors would like to thank Prof. Keith Hol-
land (University of Leeds, Leeds, U.K.) for reading
and commenting on aspects of this manuscript
and providing valued advice on scientific content.
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