Food Microbiology, 1990,7, 113-l 20

Antimicrobial activity of sodium lactate

J. C. de Wit* and F. M. Rombouts
Department of Food Science, Agricultural University, Bomenweg 2,
6703 HD Wageningen, The Netherlands
Received 28 November 1989

Sodium lactate (5%) used in neutral culture media has an antimicrobial effect which
rises above that what would be expected from its water activity lowering effect. This
antimicrobial effect was found to exist towards various lactic acid bacteria, Staphylo-
coccus aureus, and Salmonella typhimurium. It is particularly evident from increased
lag phase, decreased growth yield, and somewhat less from decreased growth rate.
Adverse growing conditions, such as suboptimum temperature, increase its anti-
microbial effect. Escherichia coli is an exception, as its growth rate is hardly affected by
5% sodium lactate, although its growth yield is lowered. The antimicrobial activity of
sodium lactate evidently reflects how various micro-organisms can cope with this
compound.

Introduction 6-3-6.7, at which more than 99.5% of

The use of sodium lactate in cooked meat
products, such as hams, sausages, pate
and turkey may offer interesting pros-
pects for shelflife extension. This seems
particularly true for products that are
handled after heat treatment. This hand-
ling, such as slicing or packaging, causes
recontamination with a flora that is diffl-
cult to control, despite nitrite, salt and
refrigeration (Debevere 1989). Moreover,
it was shown recently that sodium lactate
delays toxin production by Clostridium
botulinum in cooked-in-bag turkey prod-
ucts (Maas et al. 1989).
The efficacy of lactic acid as a meat
decontaminant is well established
(Smulders et al. 1986). However, just as
with other organic acids, the anti-
microbial activity is primarily ascribed to
the pH lowering effect and to the undisso-
ciated form of the acid, and therefore pH
dependent (Grau 1981, Gill and Newton
1982). However, meat products in which
sodium lactate is applied have a pH of
0740-0020/90/020113 + 08 $02.0010
lactic acid (pK value 3.86) is dissociated.
Sodium lactate, in a concentration of
around 2% w/w as used in meat products
(about 4% in the water phase) has a
marked effect on water activity (Chirife
and Fontan 1980). This water activity
lowering effect will result in shelflife pro-
longation, especially since these meat
products already have a fairly critical
water activity of around 0.96. The ques-
tion arises whether the shelflife pro-
longation caused by sodium lactate rises
significantly above that which would be
expected from its water activity lowering
effect.
To answer this question we have car-
ried out growth experiments with a num-
ber of spoilage and pathogenic organ-
isms. In these experiments growth in the
presence of sodium lactate was compared
to that with sodium chloride at the same
water activity value. In this way the
sodium concentration in the media with
and without lactate was the same. Also,
0 1990 Academic Press Limited
114 J. C. de Wit and F. M. Rombouts
since sodium lactate would partly replace
sodium chloride in food products, the in-
hibition by sodium lactate compared to
that by sodium chloride in media with
identical water activity (a,) could be
established. Attention was also paid to
effects of oxygen availability and tem-
perature.
Materials and Methods
Sodium lactate
Biological sodium L-lactate, 60% w/w, phar-
maceutical quality, was obtained from CCA
biochem b.v., Gorinchem, The Netherlands.
Strains, preparation of inoculum
Lactobacillus casei’ var. rhamnosus, Strepto-
coccus faecalis, Ltuconostoc mesenteroides,
Staphylococcus aureus, Escherichia coli and
Salmonella typhimurium were all from our
own culture collection.,
Inoculum was prepared by growing the
strains in Brain Heart Infusion Broth (Oxoid)
for 18-24 h at 30°C and making dilutions in
peptone physiological saline solution (PPS).
Test media were inoculated to contain
104-lo5 cells ml-‘.
Test media
Microbial growth in a medium with sodium
lactate was compared with growth in the same
medium in which sodium lactate had been
replaced by the same molar concentration of
sodium chloride. A model medium was com-
posed to approach the composition and water
activity (about 0.96) of meat products. It con-
tained per litre: 83 g meat extract (Lab Lemco
powder, Oxoidl; 83 g yeast extract powder
(Oxoid); O-9g KHzPO,; 2.6 g Na,HP0,.2HzO;
26.3 g NaCl or 83.3 g 60% w/w sodium lactate;
10 g glucose. The ingredients, except glucose
were dissolved in distilled water, adjusting the
solution to 900 ml. After sterilization (20 min,
12O”C), 100 ml of a 10% w/w glucose solution,
sterilized separately, was added. Final pH was
7.25 f O-10. For each experiment the media
originated from the same batch, differing only
in added equimolar amounts of NaCl or
sodium lactate. Therefore, the water activities
of each set of media (with NaCl or with sodium
lactate) should have been identical. The
freshly prepared media 4y values were
measured at 25°C with a Novasina EEJA-3
instrument. Sets of media o,,, measurements
did not differ more than 0.003. Of the various
batches used an average a, of 0.958 + 0.006
was measured.
Other media used were the above-
mentioned medium in which meat extract and
yeast extract were replaced by 5 g neutralized
bacteriological peptone per 1000 ml medium
(a, O-977 + 0.002, pH 7.31, and Brain Heart
Infusion Broth (Oxoid) with sodium chloride
or sodium lactate, a, 0.978 + 0.001, pH 7.3.
Because of insufficient growth in the model
medium, Lactobacillus casei var. rhamnosus
and Leuconostoc mesenteroides were tested
in MRS Broth (Gibco) with 20 g 1-l sodium
chloride, 4~ 0.967 + 0.002, and pH 6.00 *
0.005. These media were also used either with
26.3 g NaCl or with 83.3 g 60% w/w sodium
lactate 1-l.
The temperature of incubation was 3O”C,
unless otherwise stated. Aerobic cultures
were grown in 300 ml Erlenmeyer flasks with
40 ml medium, on a rotary incubator (150
rpm). With this method oxygen saturation
may not have been achieved over the entire
growth curve, but oxygen must have been
readily available during the lag phase and
early logarithmic phase of growth. For anaer-
obic growth screw-capped serum bottles were
completely filled up with freshly sterilized me-
dium and samples were taken using an injec-
tion needle. Growth was followed by plating
out adequate dilutions on Plate Count Agar
(Oxoid), using a spiral plating system (Spiral
Systems Inc., Cincinnati, OH) and counting
after 2 days incubation at 30°C. Lactobacillus
and Leuconostoc sp. were counted in MRS
Agar (Gibco; double layer technique).
Results
In Figs 1-3, growth of Streptococcus fue-
calis, Staphylococcus aureus and Salmo-
nella typhimurium in the model medium
with lactate appears to be inhibited in
comparison with growth in the same me-
dium with sodium chloride. Since both
media had exactly the same water
activity (values were close to 0.9581, the
differences in growth curves reflect the
inhibition which is specifically due to
sodium lactate, in addition to its a, lower-
ing effect. For the micro-organisms
tested there appears to be little, if any,
difference in inhibition under aerobic and
anaerobic conditions [(a) vs (b) of Figs l-
 Antimicrobial activity of sodium lactate 115

Streptococcus foecofis

o-0--o
0-O
.f

(a) (b)

2 I I I I I I
0 20 40 60 00 0 20 40 60 3
Time (h)

Fig. 1. Growth of Streptococcus fuecalis at 30°C in the model medium of pH 7.25 and a,,, O-958.
Conditions are (a) aerobic and (b) anaerobic, (0) with 5% (w/w) sodium lactate, (0) with 2.6%
(w/w) sodium chloride.

33. With all three organisms sodium lac- specific inhibitory effect of sodium lactate
tate appears to lengthen the lag phase, was studied with Streptococcus faecalis
and to diminish somewhat the maximum (Fig. 4). It appeared that the inhibitory
growth rate. Also the maximum number effect of lactate becomes more pro-
of micro-organisms in the stationary nounced with decreasing temperature.
phase is clearly lower in the lactate me- Figures 5 and 6 show that the specific
dium. inhibitory effect of sodium lactate over
The effect of temperature on the sodium chloride also exists for other lactic

Sfo~hy/ococc~.5 oureus

o-
a

0/
-J
e'
I
(a) (b)

0 20 40 60 so 0 20 40 60 00
Time (h)

Fig. 2. Growth of Staphylococcus aureus at 30°C in the model medium of pH 7.25 and a, O-958.
Conditions are as in Fig. 1.
116 J. C. de Wit and F. M. Rombouts

5Wmonello typhimurium

4- (a) (b)

I I I I I I
2
0 20 40 60
Time (h)

Fig. 3. Growth of Salmonella typhimurium at 30°C in the model medium of pH 7.25 and q.,
0.958. Conditions are as in Fig. 1.

acid bacteria. Both micro-organisms
tested, Leuconostoc mesenteroides (Fig.
5) and Lactobacillus casei var. rhamno-
sus (Fig. 6) show inhibition behaviour in
MRS medium (a, = 0.967 + O-002), simi-
lar to that observed for the organisms in
Figs l-3.
The specific lactate inhibitory effect is
relatively small for Escherichia coli, as
shown in Fig. 7. It appears that the effect

Streptococcus foecolis

21 I I I I I I
0 100 200 300 400 500 600

Time ( h )

Fig. 4. Growth of Streptococcus faecalis at various temperatures in model medium of pH 7.25

and a, 0.958. Conditions are (0) with 5% (w/w) sodium lactate, (0) with 2.6% (w/w) sodium
chloride.
 8 0-e
Antimicrobial activity of sodium lactate 117

T
2gH o/
6PJd0/.’ L euconos tot mesen teroides

c 4

20no
(a)

Time
1
0

(h)
I
20

Fig. 5. Growth of Leuconostoc mesenteroides at 30°C in MRS broth, pH 6.0 and Q 0.967.
I
40

Conditions are (a) aerobic and (b) anaerobic, (0) with 5% sodium lactate, (0) with 2.6% sodium
I
60
(b)

chloride.

of lowering a, from O-977 to 0.958 has a at suboptimum values for water activity
much more dramatic growth inhibitory and temperature. However there seems
effect, even when achieved with sodium to be an effect on maximum growth yield,
chloride. For this strain the effect of tem- as judged from the 30°C curves.
perature was also tested (Fig. 8). This
experiment essentially shows the ab- Discussion
sence of a specific lactate inhibitory effect, Weak organic acids, such as lactic acid,
towards this E. coli strain, as far as lag owe their microbial growth inhibitory
phase and growth rate is concerned, even properties to the undissociated form. The

Locfobocillus cosei’ var. rbomnosus

IO

I
oc
4 (a)

2t I I I I
0 20 40 60 60 0 20 40 60 SO

Time(h)

Fig. 6. Growth of Lactobacillus casei var. rhamnosus at 30°C in MRS broth. Conditions are as
in Fig. 5.
118 J. C. de Wit and F. M. Rombouts

Escherichio co/i

020
Time(h)

Fig. 7. Growth of Escherichiu coli at 30°C in the model medium with pH 7.25, a, 0.958, and
either 5% sodium lactate (0) or 2.6% sodium chloride (0) and in a medium with pH 7.3, a, 0.977
and either 5% sodium lactate (A) or 2.6% sodium chloride (A ).

Escherichio cob’

4 ---_
-- -*
--o-:\,
‘\\,\
-e------- ------ IOT*
2 I 1 I I I
0 loo 200 300 400 500 600
Time ( h 1

Fig. 8. Growth of Escherichia coli at different temperatures in the model medium with pH
7.25, a, 0468 and 5% sodium lactate (01, or 2.6% sodium chloride (0).
 Antimicrobial
uncharged, lipophilic form diffuses
through the cell membrane into the cell
(Warth 1986). In accordance with inter-
nal pH, dissociation takes place, resulting
in lowering of cytoplasmic pH and build-
ing up of anion concentration. Driving
forces in this diffusion process are: (i) the
concentration of undissociated acid in the
medium, which is low (0.2 mM) in the
model lactate medium tested (pH 7.21, (ii)
the permeability of the membrane. This
permeability may also be pH dependent,
as was shown by Cassio et al. (19871,
for Saccharomyces cerevisiae. They
measured that the diffusion constant for
undissociated lactic acid through the
plasma membrane increases exponen-
tially by a factor of 40 over the pH range
of 3.0-6.0. The permeability factor is
usually not considered, and to our knowl-
edge no data are available for bacterial
membranes. Increased permeability of
cellular membranes for lactic acid at
higher pH values may be an important
factor in understanding the anti-
microbial activity of sodium lactate ob-
served in neutral media. More generally,
the permeability dependency of pH may
help to explain why the antimicrobial
effect of organic acids may be less
influenced by pH changes than predic-
table from their dissociation behaviour
(cf. Eklund 1983). . A similar pumping activity is not present
In addition to accumulation of lactic
acid by diffusion, in certain micro-
organisms lactate uptake may be carrier
mediated. Carrier-mediated transport of
organic acids may be inducible, for in-
stance in certain yeasts (Cassio et al.
1987) or constitutively present if needed
for e&ix of metabolic end products such
as D,L-lactate in lactic acid bacteria and
certain Enterobacteriaceae, including E.
coli and S. typhimurium. This type of
transport is fully reversible, and the driv-
ing forces (proton gradient, lactate gradi-
ent) determine which way transport will
activity of sodium lactate 119
go (Ten Brink and Konings 1980, Ten
Brink et al. 1985).
Cellular response to accumulation of
lactate and protons may be diverse. In
general, accumulation of protons tends to
alleviate the pH gradient between cyto-
plasm and medium, which may be one of
the driving forces for uptake of certain
amino acids and other compounds
(Freese et al. 1973, Poolman et al. 1987).
Actively growing cells are able to expel
protons either by membrane-bound elec-
tron transfer and/or by the proton-
translocating ATPase complex (Ten
Brink and Konings, 1980). Accumulation
of lactate may be counteracted by carrier-
mediated efflux. Both processes will nor-
mally require energy, which cannot be
spent on synthesis and growth. This loss
of energy will result in reduced growth
rate and possibly growth yield. Cells in
the lag phase may not be able to respond
adequately due to lack of energy in the
form of proton motive force or ATP. Con-
sequently the lag phase will be prolon-
gated.
The absence of lactate inhibition in
Escherichia coli may possibly be
explained by the large surplus in proton
motive force generating power provided
by the respiration chain or the anaerobic
electron transport chain in this organism.
in Streptococcus cremoris, Leuconostoc
mesenteroides and Lactobacillus casei.
For that reason lactate should have a
stronger inhibitory effect on these lactic
acid bacteria than on E. coli (Poolman et
al. 1987).
It is clear from the experiments that
there is an antimicrobial effect of sodium
lactate, which cannot be ascribed to its
water activity lowering effect. The
specific inhibitory effect of sodium lactate
on various micro-organisms evidently
depends on how these micro-organisms
can cope with this compound. Adverse
120 J. C. de Wit and F. M. Rombouts

growing conditions, such as suboptimum trophic lactic acid bacteria (Debevere

temperature, may increase the anti-
microbial effects (Fig. 4). Escherichia coli
shows an exceptional behaviour as the
growth rate of this organism is little
affected by sodium lactate (Figs 7 and 8).
It is interesting to note that sodium lac-
tate finds its application primarily in
cooked meat products such as hams, sau-
sages, pate and turkey to extend cold
storage. The spoilage flora of these prod-
ucts consists predominantly of psychro-
1989). It appears that sodium lactate
most clearly inhibits the growth of just
these bacteria, especially under adverse
temperature conditions, such as main-
tained during cold storage. Indeed it is
often observed that the use of sodium
lactate allows 1 or 2 weeks prolongation
of cold storage of these products. Since
there are other bacteria which are less
affected by sodium lactate a shift in spoil-
age association may be expected.

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