Appl Microbiol Biotechnol (2013) 97:4691–4700
DOI 10.1007/s00253-013-4858-1
MINI-REVIEW
Production of mannosylerythritol lipids and their application
in cosmetics
Tomotake Morita & Tokuma Fukuoka &
Tomohiro Imura & Dai Kitamoto
Received: 7 February 2013 / Revised: 12 March 2013 / Accepted: 13 March 2013 / Published online: 14 April 2013
# Springer-Verlag Berlin Heidelberg 2013
Abstract Mannosylerythritol lipids (MELs) are glycolipid
biosurfactants abundantly produced by different basidiomy-
cetous yeasts such as Pseudozyma, and show not only
excellent interfacial properties but also versatile biochemical
actions. These features of MELs make their application in
new technology areas possible. Recently, the structural and
functional variety of MELs was considerably expanded by
advanced microbial screening methods. Different types of
MELs bearing different hydrophilic and hydrophobic parts
have been reported. The genes responsible for MEL biosyn-
thesis were identified, and their genetic study is now in
progress, aiming to control the chemical structure. The
excellent properties leading to practical cosmetic ingre-
dients, i.e., moisturization of dry skin, repair of damaged
hair, activation of fibroblast and papilla cells and antioxidant
and protective effects in skin cells, have been demonstrated
on the yeast glycolipid biosurfactants. In this review, the
current status of research and development on MELs,
particularly the commercial application in cosmetics, is
described.
Keywords Biosurfactant . Glycolipid . Mannosylerythritol
lipid . Yeast . Skin care . Hair care . Cell activation . Cosmetics
Introduction
Mannosylerythritol lipids (MELs), which contain 4-O-β-D-
mannopyranosyl-erythritol or 1-O-β-D-mannopyranosyl-
T. Morita : T. Fukuoka : T. Imura : D. Kitamoto (*)
Research Institute for Innovation in Sustainable Chemistry,
National Institute of Advanced Industrial Science and Technology
(AIST), Tsukuba Central 5-2, Higashi 1-1-1,
Tsukuba, Ibaraki 305-8565, Japan
e-mail: dai-kitamoto@aist.go.jp
erythritol as a hydrophilic headgroup and fatty acids as the
hydrophobic chain (Figs. 1, 2, and 3), are the functional
glycolipids abundantly produced by yeast strains of the
genus Pseudozyma (Kitamoto et al. 2002; Morita et al.
2007, 2009a). MELs exhibit not only excellent interfacial
properties as bio-based surfactants (Kitamoto et al. 1993),
but also versatile biochemical actions such as differentiation–
induction against human leukemia (Isoda et al. 1997a, b), rat
pheochromocytoma (Wakamatsu et al. 2001), and mouse
melanoma cells (Zhao et al. 1999, 2001). MELs also show
high binding affinity towards different immunoglobulins and
lectins (Im et al. 2003; Ito et al. 2007; Konishi et al. 2007;
Imura et al. 2007a, 2008). In addition, MEL-A, di-acetylated
MEL, dramatically increases the efficiency of gene transfec-
tion mediated by cationic liposomes (Inoh et al. 2001, 2004,
2010; Igarashi et al. 2006; Ueno et al. 2007). More interest-
ingly, MELs have anti-inflammatory action inhibiting the
secretion of inflammatory mediators from mast cells (Morita
et al. 2011a).
Therefore, MELs have attracted considerable interest in
recent years, besides their high biodegradability, mild
production conditions, and variety of functions. These features
of MELs would broaden their application in new technology
areas, including the food, cosmetic, and pharmaceutical indus-
tries, environmental protection and energy-saving technology
(Kitamoto et al. 2002, 2009).
Recently, we have made a breakthrough in improving the
production of MELs and in expanding the structural and
functional variety, based on advanced microbial screening
methods. One of MEL homologues is currently commer-
cially available as a new cosmetic ingredient, SurfMellow®,
by the Japanese company, Toyobo Co., Ltd. (http://
www.toyobo-global.com/seihin/cosme/surfmellow.htm).
We summarize here the latest progresses of research and
development in MELs.
4692 Appl Microbiol Biotechnol (2013) 97:4691–4700

Fig. 1 Chemical structure of CH3

mannosylerythritol lipids. a tri- O
acylated type of MEL, b di- a C
m
b c
acylated MEL, c mono-acylated CH3 O CH3 OH OH
H 2C H2C H 3C H 2C
MEL, MEL-A: R1 =Ac, R2 = H 3C
n H 3C
n H C OH
Ac; MEL-B: R1 =Ac, R2 =H; H C OH
n
OR1 O OR1 O H C OH n
OH
OH
n H C OH
MEL-C: R1 =H, R2 =Ac; MEL- O H C OH
O H C OH O O
O O O O
D: R1 =H, R2 =H CH2 CH2 HO
CH2
R2O O O R2O O O O
O

n = 4-16, m = 6-16 n = 4-16 n = 4-16

Tri-acylated MEL Di-acylated MEL Mono-acylated MEL

Structural variety of MELs
The basidiomycetous yeasts, Pseudozyma antarctica, Pseu-
dozyma aphidis, Pseudozyma rugulosa, and Pseudozyma
parantarctica produce the large amount of MELs, mainly
MEL-A (di-acetylated MEL) together with MEL-B and
MEL-C (Fig. 1), from different vegetables oils at more than
100 g/L of the production yield by intermittent feeding of
the substrates (Kitamoto et al. 1990, 1992, 2001; Rau et al.
2005a, b; Morita et al. 2006a, 2008a). Pseudozyma tsuku-
baensis produces selectively the diastereomer type of MEL-
B. The sugar moiety of the MEL-B formed by P. tsukubaen-
sis was identified to be 1-O-β-D-mannopyranosyl-erythritol
(Fig. 2), stereochemically different from the 4-O-β-D-
mannopyranosyl-erythritol of conventional MELs (Fukuoka
et al. 2008a). The new strains of P. tsukubaensis, i.e., 1D9,
1D10, 1D11, and 1E5, were recently isolated from leaves of
Perilla frutescens as the excellent producer for the diastereo-
mer type of MEL-B. Of these strains, the strain 1E5 produced
the greatest amount of the MEL-B at more than 73.1 g/L of the
production yield (Morita et al. 2010a). Pseudozyma crassa
produces the diastereomer type of MEL-A, MEL-B and
MEL-C, stereochemically different from conventional MELs
(Fukuoka et al. 2008b). Pseudozyma hubeiensis, Pseudozyma
graminicola, Pseudozyma shanxiensis, and Pseudozyma sia-
mensis were reported to produce mainly MEL-C (Fukuoka et
al. 2007a; Konishi et al. 2008; Morita et al. 2008b, c).
P. hubeiensis efficiently produces MEL-C at over 72 g/L of
the production yield by fed-batch culture (Konishi et al. 2008).
A novel MEL homologue having no acetyl groups, namely
MEL-D, was synthesized by lipase-catalyzed hydrolysis of
CH3 OH
H2C
H3C
n OH
HO C H
OR1 O H3C
n
HO C H
O O
O n
CH2
R2O
O O
n = 4-16
MEL-B (Fukuoka et al. 2011). Interestingly, MEL-D with the
conventional erythritol configuration is successfully prepared
from the conventional MEL-B, while MEL-D with the oppo-
site erythritol configuration is from the diastereomer type of
MEL-B produced by P. tsukubaensis (Fukuoka et al. 2012).
We previously reported the production of MELs with different
numbers of hydrophobic tail chain, such as tri-acylated MEL
(Morita et al. 2008a; Fukuoka et al. 2007b) from a soybean
oil-rich medium and mono-acylated MEL from a glucose-rich
medium (Fukuoka et al. 2007c).
Recently, we reported several novel MEL producers.
Pseudozyma churashimaensis, newly identified species of
the genus Pseudozyma, was isolated from sugarcane plant as
the MEL producer. P. churashimaensis OK94 produces not
only MEL-A as the main product, but also a novel type of
MEL, mono-acylated and tri-acetylated MEL, as the minor
product (Fig. 3) (Morita et al. 2011b). Ustilago scitaminea,
which is a smut fungus on sugarcane, is able to produce
selectively a large amount of MEL-B; the production yield
of reaches at 25.1 g/L from sugarcane juice (containing
19.3 % sugars) supplemented with 1 g/L of urea (Morita et al.
2011c).
Moreover, P. parantarctica JCM11752T was recently
reported to produce mannosyl–mannitol lipid possessing
mannitol (C6 sugar alcohol) as the hydrophilic part instead
of erythritol (C4 sugar alcohol) using a medium containing
olive oil and mannitol (Fig. 4) (Morita et al. 2009b). Other new
homologues possessing C5 sugar alcohol, mannosyl–arabitol
lipids and mannosyl–ribitol lipids, are also produced by
P. parantarctica JCM11752T (Fig. 4) (Morita et al. 2012).
The production and interfacial properties of these MEL homo-
logues were listed in Table 1.
H2C
H C OH
OAc
OAc
H C OH
O O
CH2
AcO O O
n = 4-16

Diastereomer type of MEL Mono-acylated, tri-acetylated MEL

Fig. 2 Chemical structure of the diastereomer type of MELs. MEL-A: Fig. 3 Chemical structure of a novel type of MEL, mono-acylated and
R1 =Ac, R2 =Ac; MEL-B: R1 = Ac, R2 =H; MEL-C: R1 =H, R2 =Ac tri-acetylated MEL
Appl Microbiol Biotechnol (2013) 97:4691–4700 4693

Fig. 4 Chemical structure of OH

the novel type of glycolipid a H 2C b OH
c
H 2C OH
biosurfactants. a mannosyl– HO C H H2C
mannitol lipid (MML), CH3 CH3 CH3
HO C H H3C H C OH HO C H
b mannosyl–arabitol lipid H 3C n H3C
n n
H C OH
(MAL), c mannosyl–ribitol OAc O H C OH n
OAc O OAc O
H C OH
n n
lipid (MRL) H C OH O H C OH H C OH
O O O O O
O O O
CH2 CH2 CH2
AcO O AcO O O AcO
O O O
n = 4-16 n = 4-16 n = 4-16

Mannosyl-mannitol lipid (MML) Mannosyl-arabitol lipid (MAL) Mannosyl-ribitol lipid (MRL)

Biosynthetic pathway of MEL
The genes responsible for MEL biosynthesis were initially
identified on a smut fungi Ustilago maydis, which produces
MELs together with cellobiose lipids at a lower concentra-
tion compared with Pseudozyma yeasts (Fig. 5a) (Spoeckner
et al. 1999). U. maydis is one of the most convenient micro-
organisms for genetic engineering due to the whole genome
information and newly developed host–vector systems
(Bölker et al. 2008). As shown in Fig. 5b, MELs would be
synthesized via the four steps of enzymatic reaction; emt1
encodes a mannosyltransferase involved in the formation of
mannosylerythritol by mannosylation of erythritol, mat1
encodes an acetyltransferase catalyzing the acetylation of
mannosylerythritol at both the C-4′ and C-6′ hydroxyl
groups of mannose, and mac1 encodes an acyltransferase
related to the acylation of mannosylerythritol (Hewald et al.
2006).
Previously, an expressed sequence tags (EST) analysis
was carried out on P. antarctica using the cells grown under
the MEL production conditions, and the genes expressed
during MEL production were tentatively categorized on the
basis of putative functions (Morita et al. 2006b).
A gene homologous to a mitochondrial ADP/ATP carrier
was the most frequent one among the genes expressing in P.
antarctica under the MEL production conditions based on
the EST analysis. The heterologous expression of the gene
by introducing a plasmid pUXV1-PaAAC1 into the yeast
cells significantly increased in MEL production (Morita et
al. 2010b). On the other hand, the expression of PaAAC1
mutant in which the conserved arginine and leucine required
for ATP transport activity were replaced with isoleucine and
serine, respectively, did not increase MEL production.
Accordingly, the contribution of PaAAC1 encoding a
mitochondrial ADP/ATP carrier to MEL biosynthesis
was genetically demonstrated for the first time. The
metabolic interaction between MEL biosynthesis and
PaAAC1, however, still remains unknown.
Moreover, among the genes obtained from the EST analysis
on P. antarctica, we identified a gene PaEMT1 encoding a
mannosyltransferase, which is an essential gene for MEL bio-
synthesis of U. maydis, on the basis of the sequence identity: a
contiguous sequence of 938 bp, PA 004, from P. antarctica
showed high sequence identity (72 %) to the gene emt1, encod-
ing an erythritol/mannose transferase of U. maydis (Morita et
al. 2010c). A gene-disrupted strain of P. antarctica, ΔPaEMT1,
failed to produce MELs, while its growth was the same as that
of the parental strain. The defect of MEL biosynthesis in a
strain, ΔPaEMT1, was complemented with a vector pUXV1-
PaEMT1 (Fig. 6).
Further investigation of the genomic analysis and
development of the gene expression system for Pseudozyma
yeasts should enable us to understand the glycolipid biosyn-
thesis, containing the gene cluster for MEL synthesis, and to
improve the production yield and selectivity of MELs.
Interfacial properties of MELs
MEL-A and MEL-B exhibit excellent surface and interfacial
tension-lowering actions and low critical micelle concentra-
tions (cmc), although the hydrophobic parts consist of main-
ly fatty acids ranging from C8 to C12. On the Wilhelmy
method, MEL-A and MEL-B show the cmc at 2.7×10−6 and
4.5×10−6 (M), respectively, and reduce the aqueous surface
tension to about 28 mN/m (Kitamoto et al. 1993).
Water-in-oil (W/O) microemulsion has attracted attention
in various fields, but the formation of microemulsions usually
requires the use of surfactant mixtures with salt or alcohol.
Recently, MEL-A was reported to form stable W/O micro-
emulsion in the ternary MEL-A/water/n-decane system, with-
out any other additives (Worakitkanchanakul et al. 2008,
2009). Dynamic light scattering and freeze-fracture electron
microscopy (FF-EM) measurements revealed that the diame-
ter of the microemulsion increases with an increase in water-
to-surfactant mole ratio (W0) ranging from 20 to 60 nm, and
the maximum W0 value was found to be 20, which is as high
as that of soybean lecithin. Thus, MEL-A has a great potential
for the formation of W/O microemulsion.
The self-assembling manner of MELs is illustrated in
Fig. 7. MEL-B and MEL-C spontaneously form giant uni-
lamellar vesicles of diameter larger than 10 μm in aqueous
solution (Kitamoto et al. 2009). It is generally difficult to
obtain giant vesicles from glycolipids, because the vesicle
 4694

Table 1 MEL producers and their products

Microbial producers Carbon sources Glycolipids Yield (g/L) cmc (M)a γcmcb (mN/m) References

Pseudozyma aphidis Soybean oil and glucose MEL-A (main), MEL-B, MEL-C 165c,d,e 1.4×10−5 26.2 Rau et al. 2005a, b; Morita et al. 2007
c
Pseudozyma antarctica Soybean oil MEL-A (main), MEL-B, MEL-C 40 2.7×10−6 28.4 Kitamoto et al. 1990, 1993; Morita et al. 2007
Soybean oil MEL-B (purified) 10 4.5×10−6 28.2 Kitamoto et al. 1990, 1993
c,d
n-Alkane MEL-A (main), MEL-B, MEL-C 140 ND ND Kitamoto et al. 2001
Glucose Mono-acylated MEL (purified) 1.3 3.6×10−4 33.8 Fukuoka et al. 2007c
Soybean oil Tri-acylated MEL (purified) ND ND ND Fukuoka et al. 2007b
c −6
Pseudozyma churashimaensis Soybean oil Mono-acylated/tri-acetylated MEL (purified) 3.8 1.7×10 29.2 Morita et al. 2011a
Pseudozyma crassa Oleic acid and glucose Diastereomer MEL-A (main), MEL-B, MEL-C 4.6c 5.2×10−6 26.5 Fukuoka et al. 2008b
Pseudozyma graminicola Soybean oil MEL-A, MEL-B, MEL-C (main) 9.6c 4.0×10−6 24.2 Morita et al. 2008b
Pseudozyma hubeiensis Soybean oil MEL-A, MEL-B, MEL-C (main) 9.6c 4.0×10−6 24.2 Morita et al. 2008b
Pseudozyma hubeiensis Soybean oil MEL-A, MEL-B, MEL-C (main) 76.3c,d,e 6.0×10−6 25.1 Konishi et al. 2008
Pseudozyma parantarctica Soybean oil MEL-A (main), MEL-B, MEL-C 106.7c,d ND ND Morita et al. 2007, 2008a
Glucose Mono-acylated MEL (purified) 1.2 ND ND Fukuoka et al. 2007c
Soybean oil Tri-acylated MEL (purified) 22.7 ND ND Morita et al. 2008a
Olive oil and mannitol MAL (purified) 18.2 2.6×10−6 24.2 Morita et al. 2009b
Olive oil and arabitol MML (purified) ND 1.5×10−6 24.2 Morita et al. 2012
Olive oil and ribitol MRL (purified) ND 1.2×10−6 23.7 Morita et al. 2012
Pseudozyma rugulosa Soybean oil and erythritol MEL-A (main), MEL-B, MEL-C 142c,d ND ND Morita et al. 2006a, b; Morita et al. 2007
Soybean oil Tri-acylated MEL (purified) ND ND ND Fukuoka et al. 2007b
Pseudozyma shanxiensis Soybean oil MEL-C 2.72 ND ND Fukuoka et al. 2007a
Pseudozyma siamensis Safflower oil MEL-B, MEL-C (main) 18.5a 4.5×10−6 30.7 Morita et al. 2008c
Pseudozyma tsukubaensis Soybean oil Diastereomer MEL-B 73.1d,e 3.1×10−6 26.1 Morita et al. 2007, 2010a; Fukuoka et al. 2012
Ustilago cynodontis Soybean oil MEL-C 1.4 ND ND Morita et al. 2008d
Ustilago maydis Sunflower oil MELs and cellobiose lipids 30f ND ND Spoeckner et al. 1999
Ustilago scitaminea Sugarcane juice MEL-B 25.1 3.7×10−6 25.2 Morita et al. 2011b

ND no data
a
cmc critical micelle concentration
b
γcmc surface tension at cmc
c
As a mixture of MELs
d
Fed-batch culture using resting cells
e
Large-scale production with a jar-fermenter
f
As a mixture of MELs and cellobiose lipids
Appl Microbiol Biotechnol (2013) 97:4691–4700
Appl Microbiol Biotechnol (2013) 97:4691–4700 4695

a Gene cluster for MEL biosynthesis

U. maydis
mac2 emt1 mac1 mmf1 mat1
Chromosome 7
um10636 um03117 um03116 um03115 um03114

b 2 Acyl-CoA
GDP-Mannose GDP CoA 2 Acetyl-CoA CoA

OH OH OH
OH
HO HO OH HO OH
O OH
OH
Emt1p Mac1p Mat1p O
O O
HO Mac2p
OH O CH3 O CH
O O O
HO HO O H3C O O
OH O O
CH3 O CH3
OH OHO H3C O O
O

Fig. 5 Genes for MEL biosynthesis. a MEL biosynthesis cluster on transferase. mat1 encodes an acetyltransferase. mmf1 encodes the pu-
chromosome 7 of Ustilago maydis. mac1 and mac2 encode the acyl- tative MEL transporter. b Presumptive biosynthetic pathway of man-
transferase responsible for MEL formation. emt1 encodes a mannosyl nosylerythritol lipids in U. maydis

structure requires strictly balanced hydrophobic and hydro-
philic groups. This indicates that the two MELs have an
excellent molecular orientation property and a superior hydro-
philic–hydrophobic balance. Interestingly, the diastereomer
type of MEL-B (Fig. 2) efficiently self-assembles into a la-
mellar (Lα) phase over remarkably wide concentration and
temperature ranges (Worakitkanchanakul et al. 2009).
In contrast, MEL-A self-assembles to form the sponge
phase (L3 phase) at concentrations above 1 mM. The sponge
phase is generally prepared from complicated multicompo-
nent systems such as surfactants with salt/co-solvent or two
oppositely charged polyelectrolytes. Interestingly, MEL-A
was demonstrated to be the first “natural” compound to
display the formation of the sponge phase by only them-
selves. The FF-TEM observation on the MEL-A assembles
clearly shows the typical morphology of the sponge phase
composed of randomly connected three-dimensional net-
work of the bilayers. In addition to the sponge phase,
MEL-A forms various liquid crystals including the bicon-
tinuous cubic phase (V2) that are clearly different from
those of MEL-B (Imura et al. 2007b). Therefore, the
difference in spontaneous curvature of self-assemblies be-
tween the two MELs, which is caused by only “one acetyl
group” on the mannose moiety, is very likely to decide
the direction of the molecular self-assembly.

a b
Ampr
Applications of MELs in cosmetics

Plasmid
ori
Neor Skin care: moisturization of dry skin
pUXV1-
MEL-A
PaEMT1
MEL-B MELs have the similar amphiphilic structure to that of
MEL-C Pgap Phsp70 ceramide-3 (Fig. 8a), which is an essential component of
the intracellular lipids of stratum corneum, and efficiently
PaEMT1
form various lyotropic liquid crystals including the lamellar
phase (Kitamoto et al. 2009). These facts imply that MELs
would exhibit ceramide-like skin care properties. We thus
S 1 2 3 initially developed in vitro assay system to estimate the
Fig. 6 An essential gene, PaEMT1, for MEL biosynthesis in Pseudozyma moisturizing property (Morita et al. 2009c). Interestingly,
antarctica. The yeast strains were grown in a medium containing glucose as the moisturizing activity of MELs was properly evaluated
the sole carbon source. a MEL formation was confirmed by TLC analysis: using a three-dimensional cultured human skin model,
the blue spots display glycolipids corresponding to MELs. Lane S MEL
standard, lane 1 parental strain, lane 2 a gene-disrupted strain, ΔPaEMT1,
TESTSKINTM (Toyobo, Japan), on the basis of the cell
lane 3 a gene-disrupted strain, ΔPaEMT1, containing a plasmid pUXV1- viability. Human skin cells were cultured and treated with
PaEMT1. b A plasmid pUXV1-PaEMT1 carrying a gene PaEMT1 sodium dodecyl sulfate (SDS) solution, and then the effects
4696 Appl Microbiol Biotechnol (2013) 97:4691–4700

MEL-A MEL-B MEL-C

CH3 OH CH3 OH CH3 OH
H2C H2C H2C
H3 C H3C H 3C
n n n
H C OH H C OH H C OH
OAc O OAc O OH O
n n n
O H C OH H C OH O H C OH
O O O O O O
O
CH2 CH2 CH2
AcO O O HO O O AcO O O

Low High
Hydrophilicity

Sponge phase (L3) Giant vesicle; Lamellar phase (L )

Fig. 7 Self-assembling manner of MELs. Each MEL (1 mM) was dissolved in water, and the self-assembled nanostructures formed were observed
by freeze-fracture transmission electron microscopy

of different lipids on the SDS-damaged cells under dry skin
conditions were examined. All MEL homologues tested
clearly expressed the recovery effect on the damaged cells
(Fig. 8b). The observed recovery effects of MELs were
comparable to those of ceramide-3.
The water retention property of MELs was further inves-
tigated on human forearm skin. The diastereomer type of
MEL-B considerably increased the stratum corneum water
content in the skin (Yamamoto et al. 2012). In addition, the
perspiration on the skin surface was clearly suppressed by
treatment with the MEL-B. These results demonstrated that
the glycolipid would show high moisturizing action by
assisting the barrier function of the skin. Therefore, MELs
would have great potential as a new and cost-effective skin

Fig. 8 Moisturizing property

of MELs. a Chemical structure
a b
of ceramide 3. b Relative
viability of the cultured skin
cells treated with SDS. The
cultured skin cells were treated SDS - + + Three-dimensional cultured
with sodium dodecyl sulfate MEL - - + human skin cells
(SDS), washed out the SDS
solution, and then re-treated with
MELs dissolved in olive oil. The 120
viability of cells was determined
Relative viability (%)

with a colorimetric method (MTT 100

assay). Ceramide was used as the
positive control. −SDS non- 80
treated with SDS, +SDS treated
60
with SDS
40

HO HN 20
O
HO 0
HO

Ceramide 3
Appl Microbiol Biotechnol (2013) 97:4691–4700
care ingredient, considering the high cost of the commercial
production of natural ceramides.
Hair care: repair of damaged hair
Ceramides present not only in the stratum corneum of the
skin but also in the cuticle of the hair, and are widely used in
skin- and hair-care compositions due to their beneficial
effects. Ceramides are also known to protect and/or repair
the hair fibers from the attack by the various agents and
treatments. Taking the moisturizing effect of MELs into
account, we investigated the hair-care properties of MELs
using damaged hair (Morita et al. 2010d). The hair samples
were washed with SDS and bleached with hydrogen peroxide
and ammonium aqueous solution, and then the effects of
different lipids on the damaged hair were examined.
On electron microscopic observation, the artificially
damaged hair showed critical cracks on the surface. Inter-
estingly, the cracks of damaged hair were repaired by treat-
ment with MEL-A and MEL-B (Fig. 9). The repairing
effects of the MELs on the damaged hair were comparable
to that of natural ceramide. In addition, the tensile strength
of the damaged hair was clearly increased, and the hair
average friction coefficient was also maintained by treat-
ment with the MELs. MELs are thus very likely to be a new
hair-care ingredient, which would be useful not only for the
recovery of damaged hair but also for providing the smooth
and flexible hair.
Activation of fibroblast and papilla cells
In cosmetic ingredients, cell activation property is a crucial point
of skin appendage morphogenesis. For instance, minoxidil,
which is the most commonly used drug for the treatment of
androgenetic alopecia, has been reported to stimulate the prolif-
eration of various skin and hair follicle cells. Of these cells, the
dermal papilla cells are known to induce follicle formation and
hair growth by transdifferentiation of an adult epidermis. Hence
the activation of the papilla cells is a key factor for the devel-
opment of a new hair growth ingredient.
Based on our previous studies on the biochemical actions
of MELs, we investigated the cell activation properties of
MELs toward cultured fibroblast and papilla cells (Morita et
Damaged hair With MEL-A
Fig. 9 The repairing effect of MELs on the damaged hair. The hair surface was observed by scanning electron microscopy. The artificial damaged
hair was treated with MEL-A, MEL-B, and ceramide dissolved in lauryl glucoside
4697
al. 2010e). As expected, MEL-A significantly increased the
viability of the fibroblast cells more than 150 % compared
with that of control cells. Moreover, the papilla cells were
dramatically activated (150 % of cell viability) with
0.001 μg/mL of MEL-A (Fig. 10). MEL-A would thus have
great potential as a new hair growth agent stimulating the
papilla cells.
Antioxidant and protective effects against oxidative
stress in fibroblast cells
In skin care cosmetics, there have been increasing interests
in the effective ingredients showing not only moisturizing
properties, but also antioxidant actions against oxidative
stress associated with the formation of reactive oxygen
species such as hydroxyl radical and superoxide anion. A
variety of antioxidants have been developed to prevent
human skin surface from oxidative injury. For instance,
unsaturated lipids like squalene are reported to function as
a scavenger of reactive oxygen species. We thus focused our
attention on the antioxidant properties of MELs, considering
that they consist of unsaturated fatty acids as the hydrophobic
part (Takahashi et al. 2012).
MELs were demonstrated to show 1,1-diphenyl-2-picryl
hydrazine radical-scavenging and superoxide anion (O2−)-
scavenging activities in a concentration-dependent manner,
but at lower levels than those of arbutin, which is well
known to act as a strong scavenger. The antioxidant prop-
erties of MELs were further examined using cultured human
skin fibroblasts under H2O2-induced oxidative stress. Inter-
estingly, MELs had a higher cytoprotective activity against
the oxidative stress compared to arbutin. On western plot
analysis, the expression of an oxidative stress marker,
cyclooxygenase-2 (COX-2), was significantly repressed in
the fibroblast cells by treatment with MELs as well as by
arbutin. Accordingly, MELs were confirmed to show anti-
oxidant and protective effects in the cells under H2O2-
induced oxidative damage, and would have potential as
anti-aging skin care ingredients.
Application to cosmetic foundation
Foundation is a skin-colored cosmetic applied to the face,
and has been used to create an even, uniform color to the
With MEL-B With ceramide
4698
Viability of papilla cells (%)
160
140
120
100
80
60
40
20
0 cial use of MELs possible.
0 0.001 0.01 0.1 1.0 10
MEL-A (µg/ml)
Fig. 10 Activation properties of MEL-A on the papilla cells. The
cultured papilla cells were incubated in a papilla cell growth medium
containing of bovine serum albumin and MEL-A, and the cells viabil-
ity was estimated by MTT assay method.
complexion, to smooth out the face and to cover spots and
flaws. Foundations containing different functional materials
have been recently developed to enhance the skin care
property. Foundation powder generally uses metal oxide
such as titanium oxide and iron oxide as its main raw
ingredients. To enhance the dispersibility and water
resistance, the particles of these metal oxides are coated
with different lipophilic materials (Fig. 11a). The coating
materials have been synthetic chemicals such as silicones,
thus the conventional foundations are difficult to exhibit
moisturizing property toward the skin.
As indicated above, MELs show unique self-assembling
properties on the different interfaces, and potential moistur-
izing property. The metal oxide particles coated with MELs
thus would have the necessary water resistance for founda-
tion as well as hydrophilicity, allowing for the development
of a novel type foundation with excellent moisture retention
property (Fig. 11b). The Japanese company, Daito Kasei
Kogyo Co., Ltd., started to provide the new powder
material.
a b
Metal oxide
particle Metal oxide
particle
Lipophilic part only
Hydrophilic part
Lipophilic part
Fig. 11 Illustration of particles of foundation. a Metal oxide particle
coated with synthetic lipophilic materials. b Metal oxidize particle
coated with MELs
Appl Microbiol Biotechnol (2013) 97:4691–4700
Further perspectives
Over the past two decade, the development of MELs has
been promoted due to the unique interfacial and biochemical
properties. As described above, the structural and functional
variety of MELs has been broadened by our developed
microbial screening methods, and this makes the commer-
100
Natural ceramides are one of the representative cosmetic
ingredients due to the excellent properties for skin and hair
care. However, the large-scale preparation of natural ceram-
ides is considerably difficult; the amount of ceramide in
each organism is very limited. Also, chemical synthesis of
isomerically pure ceramides is tedious, time-consuming, and
thus very expensive to produce for commercial applications.
In contrast, MELs are able to be efficiently produced from
inexpensive bio-resources, i.e., vegetable oil and sugarcane
juice, thus the production cost would be more decreased by
improving the fermentation processes.
There are many other potential for the practical use of
MELs besides the field of cosmetics. Details for other
applicable properties of MELs were described in our
previous reviews (Kitamoto et al. 2009). Further break-
through of understanding the biosynthetic mechanisms
and physicochemical properties of the yeast glycolipids
would allow to facilitate their commercial applications.
Acknowledgments We would like to thank to Dr. Masaru Kitagawa
and his colleagues of TOYOBO Co., Ltd. (Osaka, Japan) for providing
information on the use of MELs in cosmetics. This study was
supported by the Industrial Technology Research Grant Program
in 06A17501c from the New Energy and Industrial Technology
Development Organization (NEDO) of Japan.
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