.

Ia

Review

Antimicrobial enzymes:
Applications and future
potential in the food
industry
Claus Crone Fuglsang,Charlotte Johansen,
Stephan Christgau and JensAdler-Nissen
Antimicrobial
enzymes are ubiquitous in nature, playing a
significant role in the defense mechanisms of living organisms
against

infection

microbial

by bacteria

enzymes function

and fungi.

by degrading

Hydrolytic

anti-

key structural com-

ponents of the cell walls of bacteria and/or fungi, whereas

antimicrobial
oxidoreductases
exert their effects by the
generation

in situ of reactive

molecules.

these enzymes in food preservation

The potential

of

is still far from realized at

present.

The preservation of foods against the action of harmful

microorganisms has the dual purpose of ensuring food
safety - that is, minimizing the risk of consumers contracting a foodborne disease - and preventing or delaying microbial spoilage, which makes the food unpalatable but not necessarily unsafe. Because the same
microorganisms are rarely of concern as regards food
safety and food spoilage, effective food preservation
must generally rely on a combination of approaches.
Broadly speaking, these approaches involve:
l

minimizing the initial load of critical microorganisms

through hygienic measures;
changing the environmental conditions in the food
(temperature, pH, water activity, redox potential, etc.)
so that they become unfavourable for growth;
inactivating the living microorganisms of concern,
for example, by heat, radiation or the action of antimicrobial agents.

Claus Crone

Fuglsang

Novo

DK-2880

AIIC,

cci@novo.dk).

Denmark

Christgau

is with

207,

Department

of Biotechnology,

DK-2730

Lyngby,

01995, Else\werSc,ence Ltd

Johansen

Bagsvard,

Hovedgade
221, DK-2800

390

Stephan

and Charlotte

Herlev,

Denmark.

Nordisk

(fax: +45-4444-4096,
Osteometer

Denmark.

Technical

are with Novo

BioTech

Jens Adler-Nissen

University

of Denmark,

A/S,

e-mail:
A/S, Herlev
is at the
Building

Antimicrobial agents or preservatives are diverse in

nature, but legal, toxicological and marketing considerations have created a trend such that both the number
and amount of preservatives in use are diminishing
rather than increasing. On the other hand, the contemporary trends towards minimal processing and prolonged
shelf life of foods mean that the technological need for
effective antimicrobial agents as a tool in ensuring food
safety has not diminished, but increased.
Considering the large number of new enzymes that
are being successfully used in the processing of foods2,
it is logical to consider the potential of enzymes in the
preservation of foods. A few enzymes are already used
on a limited scale, as will become apparent, but the cost
incurred in producing them by extraction from their
natural sources is a severe limitation on their widespread use. However, with the aid of modem gene technology such enzymes could be cloned, and the prospects
of producing them cost effectively are now realistic, but
yet hardly exploited.
Cost of production is not the only parameter that has
to be considered when applying antimicrobial enzymes
in foods. Such considerations have been discussed in a
recent review on the applications of enzymes as food
antioxidants3, and the conclusions drawn are also valid
for antimicrobial enzymes. In brief, the considerations,
which are applicable for most uses of enzymes in foods,
concern the legal aspects, the potential health aspects,
the possible side effects of the activity of the enzymes in
the food and the stability of the enzymes in the food
systems. In keeping with the conclusions drawn by
Meyer and Isaksen3, antimicrobial enzymes should not
prove to be a special case compared with other types of
enzymes used in food processing. The conclusion that
the technical performance of an enzyme cannot be predicted completely, but must be investigated in realistic
food systems also holds for antimicrobial enzymes. A
few general problems that might be encountered, such
as thermostability and the potential effect on beneficial
microorganisms, will be addressed later.
The most promising candidate antimicrobial enzymes
will be discussed in this article. Two classes of enzymes
are relevant, namely the hydrolases and the oxidoreductases. The substrates for the hydrolases are key structural components of the cell walls of microorganisms;
degradation of these components inactivates the cells.
The oxidoreductases exert their effect by the generation
in situ of reactive molecules that can destroy vital proteins in the cell. Because of the great differences in the
cell-wall biochemistry of prokaryotes and eukaryotes,
the potential substrates for lytic enzymes differ for the
two groups of organisms and will therefore be discussed
separately.
Bacteriolytic enzymes
The bacterial cell wall
The cell wall, in particular the peptidoglycan component [a carbohydrate backbone consisting of p( l-4)
linked residues of N-acetyl-D-glucosamine and N-acetylmuramic acid interlinked by peptide crosslinks], is
Trends

in Food Science

& Technology

December

1995

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responsible for the rigidity of the bacterial cell.

Degradation of the cell wall may result in cell lysis
because of the inner osmotic pressure of the cell.
Peptidoglycan is present in the cell wall of both Grampositive and Gram-negative bacteria, but the molecular
structure varies from microorganism to microorganism.
The type of peptidoglycan and the cell-wall structure4
determine the resistance of the bacterial cell to the
action of lytic enzymes. The cell envelope of Gramnegative bacteria is both structurally and functionally
more complex than that of Gram-positive bacteria5. In addition to the cytoplasmic membrane, an outer membrane
(i.e. outside the cell wall), consisting of lipopolysaccharide, phospholipid, lipoprotein and protein, surrounds
the cell of Gram-negative bacteria. Because of the function of the outer membrane of Gram-negative bacteria
as a barrier to permeation, Gram-negative bacteria
appear less sensitive to bacteriolytic enzymes than
Gram-positive bacteria. Gram-negative bacteria may be
susceptible to cell-wall-degrading enzymes once the
outer membrane has been destroyed by physical or
chemical treatments6,.
Bacteriolytic enzymes are generally grouped into
three different classes8: N-acetylhexosaminidases that
catalyze the cleavage of the p( l-4)-glucosidic
linkages
in the carbohydrate backbone of the peptidoglycan,
N-acetylmuramyl-L-alanine amidases that catalyze the
cleavage between the carbohydrate moiety and the peptide moiety of the peptidoglycan, and endopeptidases
that hydrolyze the peptide bonds in the peptide
crosslinks of the peptidoglycan.

that the alkaline nature of lysozyme contributes to its

antibacterial activity9*10. Lysozymes from different
sources differ with regard to antibacterial spectrum and
specificity towards the different types of peptidoglycans. In particular, their ability to hydrolyze
U-acetylated, or otherwise substituted, peptidoglycan
varies. Lysozymes are active mainly on Gram-positive
bacteria because they lack an outer membrane. Gramnegative bacteria are more sensitive to lysozyme in
combination with EDTA6,12. Modification of lysozyme
with perillaldehyde3 (4-isopropenyl- 1-cyclohexene- lcarboxaldehyde) or conjugation of lysozyme to galactomannan14 increases the potency of the enzyme towards Gram-negative bacteria by enabling diffusion of
the enzyme across the outer membrane of the target
cell.
Lysozyme from hen egg white lyses several pathogenic and spoilage bacteria in food*,15. Although the
efficacy of lysozyme as a food preservative in various
food products has been reported (see Table I), its application in the food industry is still limited. In particular,
the enzyme is used to prevent the late gas blowing of
hard cheeses that is caused by the growth of Clostridium
tyrobutyricum9.
The efficacy of the use of lysozyme together with
other preservation techniques has been briefly studied.
Thus, the use of lysozyme in combination with other
antimicrobial enzymes, such as glucose oxidase and
lactoperoxidase20, or with traditional preservative factors,
such as sorbate, ethanol, temperature and low PH~,*,
may increase the microbial safety of food products.

Lysozyme
Lysozyme (EC 3.2.1.17), an N-acetylhexosaminidase,
lyses the cell wall of certain species of bacteria by
hydrolyzing the p( 1+4)-glucosidic
linkages of the
peptidoglycan in the cell wall. However, the antibacterial activity of lysozyme is attributable not just to its
enzyme activity, as the irreversibly inactivated enzyme
also shows antibacterial effects. It has been suggested

Other bacteriolytic enzymes

Bacteriolytic enzymes produced by various microorganisms have been reported, but there are only a few
reports on the applications of these microbial lytic
enzymes as food preservatives.
Microorganisms that produce bacteriolytic enzymes
(e.g. streptomycetes) usually express a complex of several cell-wall-degrading enzymes with different substrate

Table 1. Applications

of lysozyme

as a food preservative

Food product

Effects

Ref.

Shrimps

Total growth
with EDTA

Milkb

Inhibition

of ListerG

Codfishb

Lysozyme
microflora

suppressed
growth of 1. monocytogenes;
total growth of the natural
was inhibited by lysozyme
in combination
with EDTA

Wang

Sake

Inhibition

of sake-putrefying

Proctor

Cheeseb

Inhibition

of Clostridia

of natural

microflora

was inhibited

by lysozyme

in combination

monocytogenes

lactic

Chander

and Lewis6

Kihm et a/.6

acid bacteria

(Hiochi

bacteria)

tyrobutyricum

and Shelef

and Cunningham

Wasserfall

and Teuber*,

Fox and Gruffertylg

In all cases the enzyme

listeria

For Hiochi

Trends

was lysozyme

in milk and codfish

bacteria (lactobacihs

in Food Science

from hen egg white, and the added dosage was sufficient

and Clostridia

to inhibit growth of the bacteria mentioned

in cheese were added as inoculates

sp.)! the natural flora was inhibited

& Technology

December

1995 [Vol. 61

391

specificities, such as N-acetylmuramidase, N-acetylglucosaminidase and a number of endopeptidases22x23,

which then act synergistically to degrade the cell wall of
a given target organism. Generally, the antibacterial
activity of the isolated enzymes is lower than that of the
crude multi-enzyme complexZ4.
Fungal cell-wall-degrading enzymes
Antifungal enzymes have not attracted much attention
as potential food preservation agents. However, it is
well established that enzymes capable of degrading
fungal cell walls can be used as effective agents
against plant pathogenic fungi2s,26. The information
gathered from this field of research could be used to
assessthe applicability of this enzyme class in the food
industry.
Many of the fungi that are responsible for food
spoilage are related to fungi that cause plant diseases.
Most are deuteromycetes (Fungi Imperfecti), such as
Botrytis, Aspergillus, Fusarium and Penicillium spp.,
although ascomycetes and zygomycetes are also found
in food and feed*. Several of these food spoilage fungi
are capable of producing mycotoxins such as allatoxins,
which can cause serious health problems if ingested.

Mannans are composed of o-mannan, linked in the

l3 configuration by B( l-+4)-, B( 1+6)- and B( 1+3)linkages. Mannans can be covalently linked to polypeptide chains to form manno-proteins. Although the
role of cell-wall mannans has not been clearly defined,
it is assumed that this polymer is important for the mediation of contact between other cell-wall components, as
well as for the interaction with and attachment to
other surfaces.
Other components of the cell wall include structural
proteins, glycolipids and a wide range of special polysaccharides; although they may have important roles
in cell-cell interactions and in cell attachment, they
are unlikely targets for antifungal activities, owing
to their low abundance (generally ~5%) and complex
composition3*.
Even though there are numerous foodborne fungi, the
cell walls of the large majority share the same basic
structure that is responsible for cell-wall rigidity28; thus,
any enzymes that are active against such a structure are
potentially effective against many of the foodbome
fungi, in analogy with bacteriolytic enzymes.

Chitinases
The chitinolytic enzymes with known antifungal
The fungal cell wall
activity are mainly endochitinases (EC 3.2.1.14)
The fungal cell wall is mainly composed of polysac- although some exochitinases (EC 3.2.1.30) have also
charides, which constitute SO-90% of the total dry been shown to possess antifungal activity26.33.Numerous
weight of the wall 28.29
. A major component of many fun- chitinases from both plant and microbial sources have
gal cell walls is chitin, a polysaccharide composed of been characterized2s,26,and several have been studied for
N-acetyl-D-glucosamine linked by B( l-+4)-glucosidic
their ability to inhibit fungal growth or lyse fungal cells.
linkages. Chitin attains a highly ordered insoluble crys- Table 2 lists some of the characteristics of chitinases
talline structure comprising individual polysaccharide with known antifungal activity.
chains organized into microfibrils30. In zygomycetes,
The antifungal activity of chitinases was first discovchitin is often replaced by the deacetylated variety of ered in plants. Chitinases are produced by plants as a
the polysaccharide, known as chitosan**. Only some defense mechanism against invading fungal pathogens
species of yeast contain chitin, and then it is almost (for recent reviews see Sahai and Mannochaz6, and Bol
exclusively located in the primary septum26.
et a1.39).
Usually, a significant synergistic effect with
Chitin is associated with B-glucans and mannansO. B( 1+ 3)-glucanases is observed25,39.
B-glucans are homopolymers of D-glucose linked in the
Microbial chitinases are produced by bacteria, such as
B configuration. In most fungi, B( 1+3)- and B(1-+6)streptomycetes and bacilli, and most fungi. Bacterial
linkages dominate, and the resulting mixed-linkage poly- chitinases are secreted to assist the breakdown and
mers usually interlink individual chitin microfibrils3.
assimilation of fungal cell walls, whereas in most
I
Table 2. Fungal
Enzyme

cell-wall-degrading

source

Enzyme

, Aphanocladium
Trichoderma

with

known

type

antifungal

activity

Molecular

weight

(kDa)

Ref.

album

Endochitinase

39

Blaiseau

harzianum

Endochitinase
Exochitinase

42
40

Lorito et aLi

and LaffayjJ

Serratia

marcescens

Endochitinase

57

Jones et a1.35

Bacillus

circulans

Endochitinase

64

Roberts

Endochitinase

33

Jacobsen

et al.

Endo-P(1-13)-glucanase

78

DiPietro

ef a/.38

Hordeum
Gliocladium

vulgare

(barley)

kens

"All of the enzymes mentioned

392

enzymes

and SelitrennikofW

have been cloned

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chitinase-producing fungi, the enzymes assist fungal

cell-wall morphogenesis. Only in certain species of
mycoparasitic fungi, such as Trichoderma harzianum,
Aphanocladium album and Gliocladium virens, are
extracellular chitinases and B-glucanases used to attack
and degrade target hyphae 33,34,38.
These fungal chitinases
with antifungal activity are mostly endo-enzymes, as is
the case for plant chitinases.
Many chitinases also have bacteriolytic activity owing
to the structural similarity between the carbohydrate
backbones of peptidoglycan and chitinx6.

OZ+ B-D-glucose + H,O, + o-glucono-&lactone

(GOX catalyzed)
D-glucono-6-lactone + H,O = D-gluconic acid
(spontaneous)

The antimicrobial activity of the GOX system is due to

the cytotoxicity of the H,O, formed, although the lowering of pH by the production of D-gluconic acid may also
influence the growth of some microorganisms. The antimicrobial activity of the GOX system has been studied
in the context of food preservation with differing
ucanases
results. Tiina and Sandholm40 showed a significant
P-id
The B-glucanases with known antifungal activity are reduction in the growth of various food-poisoning bacmainly enzymes that are able to hydrolyze B(1+3) glyco- teria such as Salmonella infantis, Staphylococcus aureus
sidic linkages (EC 3.2.1.39). Studies have shown that and Clostridium perjringens. The GOX system also
B( l-3)-glucanases show a significant synergistic effect showed antibacterial activity towards both Gram-negawith chitinases on fungal cell-wall degradation. In an tive and Gram-positive bacteria in liquid whole egg4.
example with enzymes from T. harzianum, a IO-fold
However, application of the GOX system to inhibit
increase in the activity against Botrytis cinera was bacterial growth on poultry inoculated with Pseudomonas
achieved by combining purified chitinases and B-glucan- spp. and Salmonella typhimurium did not result in a
ase?. A synergistic interaction has also been observed significant decrease in the growth of these spoilage
between endo- and exo-glucanases and between endo- bacteria4*. The susceptibility of various microorganisms
B-glucanases with different specificities3.
towards the GOX system will of course be dependent on
One important concern regarding the use of B-glu- their ability to produce H,O, scavengers such as catalase
canases in food products is that most plant-derived
(EC 1.11.1.6) and/or small compounds, such as glumaterial contains various B-glucans. As B-glucans may tathione and ascorbic acid, which are readily oxidized
be important for the maintenance of organoleptic prop- by H,O, and also by the more reactive hydroxyl radical
erties such as the texture, viscosity and appearance of derived from the spontaneous reaction of H,O, with
plant cell-wall-containing foods, the addition of exogenous metal ions. The GOX system is probably more likely to
B-glucanases might adversely affect these properties.
be applied in food processing than as a shelf-life preservation agent, because long-term exposure of a food
Other fungal cell-wall-degrading enzymes
product to H,O, could cause rancidity of the food as a
Because chitin and B-glucans comprise the majority
result of lipid oxidation.
of the cell wall in most fungi, the value of additional
The GOX system is not commercially applied to preenzymes as antifungal agents is difficult to assess. vent microbial spoilage; however, it is used in combiHowever, the structural components of the cell walls of nation with catalase as an antioxidant in food prodsome fungi and most yeasts also include other types of ucts, such as salad dressings, because the net reaction
polysaccharides (e.g. mannans, o-glucan and cellu- from the combined system
results in the removal of 0,
lose)27.Thus, enzymes such as mannanase and o-glucan(B-D-glucose + %O, + D-gluconic acid)43. The GOXases that act on these structures might act as antifungal
catalase system may therefore indirectly affect the
agents against such fungi, although the antifungal activ- growth of strictly aerobic microorganisms owing to 0,
ity of these enzymes remains to be investigated.
depletion. Some reports have shown a decrease in
microbial growth upon GOX-catalase treatmenta, but it
Antimicrobial oxidoreductase systems
is unclear whether this was the result of 0, depletion.
Unlike the lytic enzymes, oxidoreductase enzymes do
not possess antimicrobial activity themselves. However,
Lactoperoxidase
Lactoperoxidase (LP; EC 1.11.1.7) is a 70-80 kDa
reaction products from reactions that are catalyzed by a
given antimicrobial oxidoreductase system are cytotoxic
porphyrin-containing peroxidase that is secreted from
and exhibit antimicrobial activity. The most-studied oxi- various mammalian glands45; thus, LP is found in, for
doreductase systems in the context of food preservation example, milk and saliva. LP catalyzes the oxidization
are the glucose oxidase and lactoperoxidase systems.
of thiocyanate (SCN-, a widely distributed compound in
animal tissue and secretions45) to hypothiocyanate
Glucose oxidase
(OSCN-) or to (SCN), by using H,O, (see Box 1). The
The glucose oxidases (GOX; EC 1.1.3.4) produced by LP systems (involving LP, H,O, and SCN-) have long
molds such as Aspergillus niger and Penicillium spp. been known to be one of the major contributors to the
catalyze the formation of H,O, and D-glucono-S-lactone
antibacterial activity of milk. Bovine LP is fairly thermo(which then reacts with H,O to form D-gluconic acid) stable (tolerates short-time pasteurization at more than
from 0, and B-D-glucose in a two-electron transfer 60C)45, but is sensitive to oxidation by H,O, if SCNreaction:
concentrations are low. The enzymatically produced

Trends

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393

Box 1. Antimicrobial activity of lactoperoxidase: possible

mechanisms
Reactions catalyzed by lactoperoxidase?
H,O, t SCN + OSCN- t H,O
or
ZSCN- t H,O, t 2H+ -+ (SCN), t 2H,O
(SCN), t H,O --f OSCN- t SCN- t 2H
Generation of higher oxyacids:
HOSCN t H,O, 3 HO,SCN t H,O
HO,SCN t H,O, 4 HO,SCN t H,O
Oxidation of protein sulfhydryls4:
Protein-SH t OSCN- + Protein-S-SCN t OK
Protein-S-SCN t H,O + Protein-S-OH + SCN- + H+

OSCN- or higher oxyacids are responsible for the antimicrobial activity48*49, presumably because they react
with protein sulfhydryls (see Box 1)47, causing inactivation of enzymes, especially enzymes that depend on
an active-site cysteine residue for their activity; however, this is probably not enough to explain the severe
effect of the LP system on some microorganisms. The
LP system should probably be considered a more effective antimicrobial system than the H,O,-generating
GOX system or H,O, itself 50,51,owing to the higher antimicrobial potency of OSCN- or the higher oxyacids. As
LP has a pH optimum in the slightly acidic range
(pH 5)51, the system functions optimally at this pH. The
possibility that HOSCN might diffuse through the cell
membrane of the target microorganism and cause damage intracellularly could contribute to the increased
antibacterial activity at low pH.
Several excellent reviews have been written on the
occurrence of, and antibacterial properties of, the LP
system 45*48;in particular, the bacteriostatic and bacteriotidal properties of the LP system in milk have been
extensively studied. At low cell concentrafions the LP
system is bacteriocidal against several Gram-negative
bacteria, including Escherichia coli, Pseudomonas aeruginosa and S. typhimurium5, whereas at higher cell numbers Gram-negative bacteria seem to resume growth
after a lag phase5. The LP system shows bacteriostatic
and in some cases bacteriocidal activity towards Grampositive bacteria. The antibacterial activity is highly
dependent not only on the pH and the relative amount of
available substrate but also on the growth phase of the
target organisms and the media in which the system is
tested. Protein-rich media that contain oxidizable compounds will scavenge the activity of the LP system. The
inhibition by the LP system of psychrotrophic bacteria
and S. typhimurium on poultry has been studied53; a significant effect (a reduction of up to 80% in colony-forming units of S. typhimurium) was observed. The effect
was highly dependent on the time of exposure and the
temperature at which the treatment was carried out.
The natural occurrence of LP in milk can be utilized
in the preservation of dairy products. The antibacterial
activity of LP in milk is enhanced if SCN- and an oxidase
394

system, such as glucose oxidase, for H,O, production

are addeds1aS4.Some guidelines for the use of the
LP system for the preservation of milk are given in
a document produced by the International Dairy
Federation55. This concept cannot be applied to other
foods, in which LP is not naturally present. Thus, the LP
system will have to be supplemented; moreover, LP is
currently not produced on an industrial scale at a costcompetitive price. However, the concept has been
applied in dental care for enhancing the antibacterial
activity of naturally occurring LP in human saliva. By
adding GOX and SCN- to toothpaste, the LP system is
believed to inhibit growth of oral streptococci56.
Other antimicrobial oxidoreductase systems
Myeloperoxidase from human leukocytes is known to
be one of the components in the human defense system
active against invasive pathogenic microorganisms.
Myeloperoxidase catalyzes the oxidation of Cl- to
hypochlorite (OC1m)57:
Cl- + H,O, + OCl- + H,O
Like OSCN-, OCl- is a much more potent antimicrobial
compound than H,O,, probably owing to a more specific
oxidation of proteins and/or enzymes, and also to the
formation of chloroamines53. This offers the opportunity
for a wider spectrum of microorganisms to be affected
at lower enzyme and substrate concentrations than is
possible with the GOX system.
Microbial enzymes with activities similar to that of
the myeloperoxidase do exist. Caldariomyces fitmago
produces a heme-containing haloperoxidase that shows
antimicrobial activity against a range of different
microorganisms 58. The vanadium-dependent haloperoxidase from Curvularia inequalis has been shown to produce HOC159; however, its antimicrobial properties have
not yet been reported. The fact that these enzymes are
produced by microorganisms makes them interesting
from the point of view of commercialization, as recombinant expression would enable their production on an
industrial scale.
Antimicrobial enzymes - present and future
Although antimicrobial enzymes represent a natural
alternative to traditional preservation agents, their use is
limited because of their low cost-efficiency.
Bacteriolytic enzymes offer protection against only a
few bacteria, owing to their specificity. Furthermore, antimicrobial activity should be achieved using reasonable
enzyme concentrations - a general concern. So far, the
fungal cell-wall-degrading enzymes have not been studied in the context of food preservation, and therefore no
major conclusions should be drawn before appropriate
studies have been conducted. The oxidoreductase systems may affect a broader spectrum of microorganisms,
but suffer from the drawback of being dependent on
the availability of substrate for the generation of antimicrobial reaction products. Also, the effect can be scavenged in food systems in which oxidizable compounds
Trends in Food Science & Technology December 1995 [Vol. 61

are abundant. The combination of antimicrobial enzymes, such as lytic enzymes with different specificities,
or the combination of antimicrobial enzymes with other
preservation methods, such as temperature treatment,
may broaden their target range and increase their use in
the future, but few studies have been carried out yet on
the effects of such combinations.
So far, the only antimicrobial enzymes to be produced
in large quantities at a cost-competitive price are lysozyme from hen egg white and GOX. These are the only
antimicrobial enzymes that are applied in the food
industry today (GOX is applied in combination with
catalase). However, heterologous expression of enzyme
genes in microorganisms on a large scale might in the
future provide new antimicrobial enzymes that can be
readily purified and used on an industrial scale for the
food industry. Until now, attempts to express mammalian or plant enzymes recombinantly have had limited success. Thus, the use of LP is probably limited to
food systems in which it occurs naturally, such as dairy
products, as mentioned earlier. Because heterologous
expression of microbial enzymes is much more readily
achieved, it seems logical to look for microbial alternatives to LP. Product approval for the use of recombinantly produced enzymes in foods can be problematic in
Europe, while in the USA, enzymes derived from genetically manipulated microorganisms are regulated by
the US Food and Drug Administration, along with all
other food additives. The reluctance of consumers to
accept food products containing such enzymes should
not be underestimated.
It is unlikely that the ingestion of antimicrobial
enzymes would present any special health problems
(obviously the LP system in milk and hen egg white lysozyme do not). Like most other enzymes, they will be degraded by the human digestive system before they reach
the large intestine, where they could possibly interact with
the natural microbial flora. However, this assumption
is based on speculation, because no studies appear to
have been carried out in an effort to elucidate this issue.
Another approach to the introduction of antimicrobial
enzymes in the food industry and in food products is the
direct use of transgenic enzyme-producing, food-grade
microorganisms in the food industry. This approach is
not generally possible, only in special cases where the
food product either already contains microorganisms or
is processed with the aid of microorganisms (i.e. wine,
beer, soy products, cheese, etc.). In order to pursue this
strategy, the processing conditions have to be developed
such that they preserve enzyme activity at all times a general concern for the application of antimicrobial
enzymes in the food industry.
In order to obtain benefit from antimicrobial enzymes,
it is of course necessary to preserve enzyme activity.
Like all other enzymatic systems, activity will be dependent on pH, temperature, the presence of salts, etc.
Another important aspect is enzyme stability (pH and
thermostability) if the enzymes are added before processing. High thermostability is a prerequisite of an
enzymatic system if it is to be used in combination with
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heat treatments. However, there are a large number of

food systems that are not preserved by heat, such as
cured or pickled meat and fish products. In such products, Listeria monocytogenes may present a health hazard; thus, there is a need for new preservation principles
to supplement the traditional preservation techniques of
curing and salting. In these cases, the activity of the
enzymes at low water activity will be of more relevance
than thermostability. Related to cured products are those
products that are preserved by fermentation, including
sour-milk products. Antibacterial enzymes are hardly
relevant for such products because their low pH is a
very effective antibacterial principle in itself, and
because the addition of enzymes may interfere with the
growth of the starter culture. Antifungal enzymes might,
however, be of interest for use on the surface of, for
example, hard cheeses.
Scientifically, the most urgent problem is that there is
so little understanding of the effectiveness of the use of
antimicrobial enzymes in conjunction with the other
components of food preservation systems. Systematic
studies on such complete preservation systems may
show promising synergistic effects between antimicrobial enzymes and the classical preservation agents
or methods. Specific, cost-effective uses of the antimicrobial enzymes might result from such investigations.
References
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T. (1994)

Adler-Nissen,
Meyer,

Trends Food SC;. Technol.

J. (1987) Trends Biotechnol.

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Next year in TIFS

396

The following

are just some of the articles commissioned

information

resources for food scientists and technologists

Internet

Pervaporation

Time-temperature

Controlling

Dye-containing

media for isolating,

Flavour

to food proteins

Improving

our ability

to assess the potential

Molecular

imprinting

technology

On-line

Preventing

Minimal

The metabolic

Natural

Advances

The sensory perception

Competitive

for aroma recovery

integrators

lipid oxidation

binding

applications

detecting

in controlling

Microbial

production

In-line

viscosity

of food flavours
in research and QC

Databanks

Project

for fruit and vegetables

The multiple

microorganisms

measurements

Food Intolerance

plants

of food flavour,

foodborne

risk of food carcinogens

food freezing

The role of fat in air stabilization

The future of MALDITOF

The European

from tropical

fluid extraction

and enumerating

The microbial

fate of resistant starch

exclusion

and edible film technology

food additives

NMR spectroscopy/imaging

Trends in supercritical

of image analysis
l

On-line

in food emulsions

food allergy
processing

and more

for TIFS in 1996:

effects of antioxidant

Applications

Understanding

of enzyme

vitamins

infusion

bulk solids mixing

texture and mouthfeel

adhesion/detachment

Trends

on food-contact

in Food Science

& Technology

surfaces

December

1995 [Vol. 61