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Responses of Lactic Acid Bacteria To o X y G e N
Responses of Lactic Acid Bacteria To o X y G e N
Published by Elsevier
FER 0073S
Seamus Condon
1. S U M M A R Y 2. I N T R O D U C T I O N
A small number of flavoprotein oxidase en- In this paper the term Lactic Acid Bacteria
zymes are responsible for the direct interaction of (LAB) is interpreted in the traditional way i.e. the
lactic acid bacteria (LAB) with oxygen; hydrogen four genera Streptococcus ( S.), Leuconostoc
peroxide or water are produced in these reactions. ( Leuc.), Pediococcus ( P.) and Lactobacillus ( L.).
In some cultures exposed to oxygen, hydrogen The group includes some strict anaerobes but the
peroxide accumulates to inhibitory levels. majority of strains investigated are aerotolerant to
Through these oxidase enzymes and N A D H some degree, and often completely so. With very
peroxidase, 02 and H202 can accept electrons few exceptions, LAB react to 02 and the conse-
from sugar metabolism, and thus have a sparing quences of such reactions may be beneficial or
effect on the use of metabolic intermediates, such detrimental. LAB generally grow satisfactorily in
as pyruvate or acetaldehyde, as electron acceptors. the absence of 02 , and in its presence some are
Consequently, sugar metabolism in aerated cul- inhibited partially or completely. Consequently, it
tures of LAB can be substantially different from is quite usual to consider that the normal growth
that in unaerated cultures. Energy and biomass metabolism of LAB is anaerobic and that
yields, end-products of sugar metabolism and the metabolism in the presence of O 2 is somewhat
range of substrates which can be metabolised are aberrant.
affected.
Lactic acid bacteria exhibit an inducible oxida-
tive stress response when exposed to sublethal 3. E N Z Y M A T I C R E A C T I O N S OF LACTIC
levels of H202. This response protects them if ACID BACTERIA WITH OXYGEN
they are subsequently exposed to lethal concentra-
tions of H202. The effect appears to be related to In general, LAB remove O z from solution, often
other stress responses such as heat-shock and is at substantial rates when their environment con-
similar, in some but not all respects, to that previ- tains a substrate which they can oxidise. The
ously reported for enteric bacteria. oxidation-reduction reactions involve a transfer
of 1, 2 or 4 electrons to the dioxygen molecule as
* Presented at the Second Symposium on Lactic Acid Bacteria follows:
- - Genetics, Metabolism and Applications, 22-25 Septem-
ber 1987, Wageningen, The Netherlands. 02 + l e - ~ O 2
02 + 2 e - + 2H + --* H 2 0 2
Correspondence to: S. Condon, Department of Food Microbi-
ology, University College, Cork, Republic of Ireland. O 2 + 4 e - + 4H + ~ 2 H 2 0
Table 1
Reactions involving molecular oxygen or oxygen metabolites catalysed by enzymes of lactic acid bacteria
5. 2 0 2 + 2 H + super°xidedismutase)H202 + 0 2
6. N A D H + H + + H 202 NADH peroxidase)2H 20
ring naturally or constructed by mutation) form The EMP pathway of sugar metabolism is used
lactate, ethanol and CO 2 in the presence or ab- by streptococci, pediococci and some lactobacilli.
sence of 02 [16]. The ethanol branch of the hetero- In unaerated (not necessarily strictly anaerobic)
lactic (phosphoketolase) pathway is needed to re- cultures, with excess glucose as substrate, most of
generate NAD + for dehydrogenation of glucose- the pyruvate is converted to lactate to reoxidise
6-phosphate and 6-phosphogluconate. The regu- NADH. Many of these bacteria have the genetic
lation of synthesis of the enzymes concerned with capacity to make products other than lactate from
the alternative routes of acetylphosphate metabo- pyruvate, such as acetate, CO2, formate, ethanol,
lism is ideally organised to take advantage of the acetoin, diacetyl and 2,3-butanediol [see Figs. 2-4]
presence or absence of 02. When O2 is available and their accumulation can be demonstrated by
the specific activities of N A D H oxidase and varying the conditions of sugar utilisation. For
acetate kinase are high and those of phosphate example, several lactic streptococci accumulate
acetyl transferase and alcohol dehydrogenase are substantial amounts of formate, ethanol and
low, thus facilitating acetate synthesis and high acetate and correspondingly less lactate when
energy phosphate conservation. In the absence of grown in glucose- or lactose-limited unaerated
O 2 the leuconostocs respond with greater amounts chemostat cultures, whereas with excess sugar the
of phosphate acetyl transferase and alcohol dehy- same cultures are homolactic. The production of
drogenase and less N A D H oxidase and acetate products other than lactate in the sugar-limited
kinase activities [16]. cultures is attributed to relatively low intracellular
The switch from ethanol to acetate synthesis levels of fructose-l,6-bisphosphate (FBP), an
doubles the amount of ATP that is formed from essential activator of lactate dehydrogenase in
274
these strains [60]. The presence of 0 2 often has ing and FBP levels were low. However, some oral
significant effects on the metabolism of pyruvate streptococci accumulate these end-products even
in these and other LAB. when glucose is in excess in strictly anaerobic
conditions. Washed cell suspensions of anaerobic
5.2.1. Pyruvate to lactate transformation glucose cultures of S. mutans accumulated sub-
Accumulation of products other than lactate stantial amounts of formate, acetate and ethanol,
from sugars by homofermentative LAB requires as well as lactate, under strictly anaerobic condi-
an alternative mechanism to lactate dehydro- tions from excess glucose or galactose. The fer-
genase (LDH) to oxidise NADH, or an alternative mentation became much more homolactic in the
P /~P AlP
Pyruv~e b~T~p~
~ Pyru~te• ~ TPPN ,- AcetYl~hate - ~ J' , ~:emte
02 %o2 c~
Fig. 3. Pyruvateoxidasepathway.
less, C4 compounds have been observed occasion- metabolise substrates such as glycerol, mannitol,
ally in cultures of some streptococci growing in a sorbitol and lactate that they do not utilise in the
sugar-based medium without additional pyruvate absence of O 2 [reviewed in 31]. In our laboratory,
[41,73,79,84,85]. Since two molecules of pyruvate the Oz-dependent utilisation of lactate by L.
are needed in such cultures to make one C4 com- plantarum has been of particular interest [53,89].
pound, oxidation of two N A D H molecules is a The probable pathway is via dehydrogenation of
prerequisite for synthesis. lactate to pyruvate followed by oxidation to acetyl
Accumulation is, therefore, more likely in the phosphate with pyruvate oxidase and finally con-
presence rather than the absence of 02. A survey version of acetyl phosphate to acetate with synthe-
55 ° C). Unlike the enteric bacteria, the protective its effect on their growth and metabolism. Appl. Micro-
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