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Antimicrobial
And Spermicidal
Activity Of Native
And Recombinant
Pediocin CP2:
A Comparative
Evaluation
2012
ARCHIVES OF CLINICAL MICROBIOLOGY
Correspondence:
balvirkumarbt@gmail.com
balvirkumar@yahoomail.co.in
Abstract
Title: Comparative analysis of antimicrobial and spermicidal activity of native and
rec-pediocin CP2
Methods and Findings: Herein, we report studies carried out to comparatively investigate antimicrobial activity of native and recombinant pediocin CP2
against general human pathogens and opportunistic pathogens associated with
BV, gastrointestinal infections and others. Further, we demonstrate the spermicidal
activity of pediocins, on human sperm so that it could be considered as a potent
spermicidal agent.
Conclusions: Pediocins have the ability to target a relatively wide range of pathogenic bacteria which is an important advantage, especially to replace antibiotics
with much safer therapeutics and to combat the ever growing problem of antibiotic resistance, when compared to other antibiotics. They have the potential to
perform a very specific role like treatment of bacterial vaginosis, gut infections
and peptic ulcers. Preliminary experiments with rec-pediocin CP2 have proved its
better effectiveness at fighting Bacteroides, Candida, Escherichia, Enterococcus,
Helicobacter, Gardnerella, Klebsiella, Listeria, Neisseria, Propionibacterium, Staphy
lococci, Streptococci and Vibrio infections compared to native pediocin CP2. Study
has also established pediocins as potent spermicidal agents with very impressive
market value in the form of personal care products.
Keywords: Pediococcus acidilactici, Pediocin CP2, Recombinant pediocin, Antimicrobial peptides, Spermicidal activity
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Introduction
Pediocin CP2 was initially purified and characterized from
Pediococcus acidilactici MTCC 5101 [1-4]. It possesses a
wide spectrum of antimicrobial activity against Aspergillus
flavus, Clostridium sporogenes, Lactobacillus brevis, L. bul
garicus, Leuconostoc mesenteroides, Listeria monocytoge
nes, Micrococcus flavus, Neisseria mucosa, Pediococcus aci
dilactici, P. pentosaceus, Pseudomonas aeruginosa, P. putida,
Staphylococcus albus, S. aureus, Streptococcus mutans and
S. pyogenes, that raised the possibility of its potential as an
ingredient in medical and/or personal care products. Later
on, it was genetically manipulated and its synthetic fusion
gene construct was cloned and expressed in heterologous E.
coli BL21(DE3) using pET32(b) expression vector. It was engineered with the aim to improve its antimicrobial spectrum,
enhance its production and facilitate its purification from
cell lysates[5]. Parental N-terminal sequence of pediocin CP2
(KYYGNGVTCGKHSC) was changed to TKYYGNGVSCTKSGC
as it could improve its biopreservative potential and effectiveness against sour-spoilage of dairy products [6]. Rec-pediocin
expression was induced by adding IPTG to early log phase
cultures of E. coli BL21(DE3) transformed with recombinant
pET32(b)-pedA. Rec-pediocin was purified from cell lysates by
employing two tandem affinity chromatographic steps with
an intermediate enterokinase cleavage step. Highly pure recpediocin was recovered from 2nd affinity column and further
characterized biochemically as well as biologically for its antimicrobial spectrum [5].
BV is a pretty common condition of vagina that occurs when
natural balance of the vaginal microflora gets disturbed due
to infection by bacteria, fungi or Trichomonas. Numerous reports across the world have shown its association with anaerobes like Bacteroides spp. (Prevotella spp.), Escherichia coli,
Gardnerella vaginalis, Mobiluncus spp., Mycoplasma hominis,
Peptostreptococcus spp., Staphylococci, Streptococci, and/or
viruses [7-11]. BV is asymptomatic in approximately one-half
of all cases, but is associated with a wide variety of symptoms
including foul odor, presence of clue cells and an increase
in pH of the vagina to >4.5 [12-13]. BV can cause adverse
outcomes of pregnancy, including preterm delivery, premature labor, premature birth, infection of the amniotic fluid,
infection of the uterus after delivery, death of the fetus or
new born and induction of pelvic inflammatory disorder [10,
14-16]. BV enhances susceptibility to infections by HIV [17]
and HSV type 2 [8] as it directly causes an increase in the
rate of viral replication and disease progression [18-19]. Toxins released by BV-associated microorganisms may cross the
placenta and may cause permanent brain damages leading to
development of Parkinsons disease and schizophrenia in the
affected child [20-22]. BV is also responsible for recurrence of
UTIs. G. vaginalis may assume a pathogenic role in males too
as infection can extend up to the prostate or bladder especially in immuno-compromised patients who have undergone
a urological procedure [23-25].
Currently, preventive therapies for BV rely almost exclusively
on the use of antibiotics such as clindamycin and metronidazole administered either orally or intravaginally [26] and gives
initial cure rates of approximately 60-90% [27]. However, antibiotic therapy has been reported to impose several side effects such as diarrhea, dizziness, headache, loss of appetite,
nausea or vomiting, stomach pain or cramps [28]. In vitro experiments established the fact that antibiotics clindamycin and
metronidazole could inhibit healthy vaginal Lactobacillus spp.
at concentrations lower than doses topically recommended
for treatment [29-30]. Moreover, emergence of multi-drug
resistant phenotype in BV associated pathogenic bacteria; it
has become imperative to develop alternative therapeutics/
prophylactic measures against these pathogens [31-34].
Till date, nisin is the only bacteriocin that enjoys GRAS (Generally Recognized As Safe) food additive status given by the
Food and Drug Administration (FDA). It was also shown to
have impressive spermicidal activity [35]. An earlier study by
Sutyak et al. [36] that convincingly demonstrated the efficacy of antimicrobial peptide subtilosin to inhibit growth of
G. vaginalis and established its spermicidal activity, is also
of particular importance as this makes it a highly recommendable compound for inclusion in topical BV treatments
and human contraceptive products. They conducted tissue
sensitivity assays using human ectocervical tissue model and
indicated its safety for human use. Subtilosin was shown to
eliminate motility and forward progression of human spermatozoa in a concentration-dependent manner. Results of the
previous studies on other LAB bacteriocins therefore suggest
that antimicrobial peptides (AMPs), including native and recombinant pediocin CP2, may enhance current therapeutics/
prophylactic measures against BV as well as contraception as
they also possess both spermicidal and antimicrobial properties, prompting our investigation.
The human gastrointestinal tract constitute a complex and
diverse ecosystem of microbiota or commensal microflora inhabiting approx. 2,000 different species of aerobic, facultative
and anaerobic bacteria, the majority of which reside in the
intestines [37]. The diversity of gut microbiota is dependent
upon the age, diet and health status of the individual [38].
Intestinal microbiota may also play a critical role in disease
suppression and maintaining gut homeostasis [39-40]. Some
forms of cancers such as gastric cancer [41] and colon cancer [42] are also associated with the bacterial pathogenesis.
Probiotics have been recommended as GRAS biotherapeutic
agents for prevention and cure of human gastrointestinal diseases [37]. Present study is an attempt to unleash the antimi Under License of Creative Commons Attribution 3.0 License
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TABLE 1. Growth conditions of indicator microorganisms used to study antimicrobial spectrum of pediocins.
Microorganism
Gram
Nature
Growth
Medium
Nature
Temp. (oC)
/pH
Inc. Time
S.P.
(Days)
Fungus
PDA
Aerobe
28 / 5.5
3 days
30
Fungus
PDA
Aerobe
28 / 5.5
3 days
30
+ ve
NB
Aerobe
37 / 7.4
24 h
15
- ve
RCB
Anaerobe
37 / 6.8
5 days
30
- ve
RCB
Anaerobe
37 / 6.8
48 h
30
- ve
CMM
Anaerobe
37 / 7.2
72 h
30
Yeast
YEPD
Aerobe
30 / 7.2
48 h
60
Yeast
YEPD
Aerobe
30 / 7.2
48 h
60
+ ve
RCB
Anaerobe
37 / 6.8
48 h
30
+ ve
RCB
Anaerobe
37 / 6.8
48 h
30
- ve
LB
F. anaerobe
37 / 7.2
24 h
30
- ve
LB
F. anaerobe
37 / 7.2
24 h
30
- ve
LB
F. anaerobe
37 / 7.2
24 h
30
- ve
LB
F. anaerobe
37 / 7.2
24 h
30
+ ve
MRS
F. anaerobe
37 / 6.5
24 h
30
+ ve
MRS
F. anaerobe
37 / 6.5
24 h
30
+ ve
MRS
F. anaerobe
37 / 6.5
24 h
30
- ve
BHI
Anaerobe
37 / 7.4
48 h
30
+ ve
CB
Anaerobe
37 / 7.2
48 h
30
- ve
NB
F. anaerobe
37 / 7.2
24 h
30
- ve
NB
F. anaerobe
30 / 7.2
24 h
15
+ ve
MRS
F. anaerobe
30 / 7.4
24 h
30
+ ve
MRS
F. anaerobe
37 / 6.5
24 h
30
+ ve
MRS
F. anaerobe
37 / 6.5
24 h
30
+ ve
MRS
F. anaerobe
37 / 6.5
24 h
30
+ ve
MRS
F. anaerobe
37 / 6.5
24 h
30
+ ve
MRS
F. anaerobe
37 / 6.5
24 h
30
+ ve
MRS
F. anaerobe
37 / 6.5
24 h
30
+ ve
MRS
F. anaerobe
37 / 6.5
24 h
30
+ ve
TSY
Aerobe
20 / 6.5
24 h
30
+ ve
MRS
Aerobe
25 / 6.5
48 h
30
+ ve
BHI
Aerobe
37 / 7.4
24 h
30
+ ve
NB
Aerobe
30 / 7.4
48 h
90
- ve
NB
Aerobe
37 / 7.4
24 h
30
- ve
NB
Aerobe
37 / 7.4
24 h
30
2012
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+ ve
MRS
Aerobe
30 / 6.5
24 h
30
+ ve
MRS
Aerobe
37 / 6.5
24 h
30
+ ve
MRS
Aerobe
37 / 6.5
24 h
30
P. pentosaceus NCDC 35
+ ve
MRS
Aerobe
37 / 6.5
24 h
30
- ve
NB
Aerobe
30 / 7.4
24 h
30
- ve
NB
Aerobe
30 / 7.4
24 h
30
- ve
NB
Aerobe
30 / 7.4
24 h
30
+ ve
BHI
Anaerobe
25 / 7.4
5 days
30
+ ve
BHI
Anaerobe
25 / 7.4
5 days
30
- ve
NB
F. anaerobe
37 / 7.2
24 h
30
- ve
BHI
F. anaerobe
37 / 7.4
24 h
15
- ve
BHI
Aerobe
37 / 7.4
24 h
15
+ ve
BHI
Anaerobe
25 / 7.4
5 days
30
+ ve
BHI
Anaerobe
37 / 7.4
24 h
15
+ ve
BHI
Anaerobe
37 / 7.4
24 h
30
S. epidermidis MTCC435
+ ve
NB
Anaerobe
37 / 7.4
24 h
30
+ ve
MRS
Aerobe
37 / 6.5
48 h
30
+ ve
MRS
Aerobe
37 / 6.5
48 h
30
+ ve
MRS
Aerobe
37 / 6.5
48 h
30
+ ve
BHI
Aerobe
37/ 7.4
48 h
30
+ ve
MRS
Aerobe
40 / 6.5
24 h
30
- ve
NB
Aerobe
37 / 7.4
24 h
15
- ve
BHI
F. anaerobe
30 / 7.2
12 h
30
F. anaerobe- Facultative anaerobe, Inc. Time- incubation time; S.P. (Days)- subculturing period
a
b
Results
In this study the antimicrobial properties and spermicidal
properties of native and rec-pediocin CP2 of P. acidilactici
MTCC 5101 were comparatively evaluated.
2012
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Rec-pediocin
CP2
-*
10
12
10
12
11
14
16
19
19
21
14
15
20
21
20
21
13
18
18
22
14
15
15
23
20
21
16
18
18
19
11
13
21
23
16
18
25
27
L. monocytogenes MTCC
657
16
22
18
20
18
22
16
23
P. acidilactici LB 42
14
23
12
13
P. pentosaceus NCDC 35
14
15
13
15
12
13
10
11
13
15
21
23
12
13
12
16
13
16
S. epidermidis MTCC435
S. mutans MTCC890
13
16
15
17
19
21
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FIGURE 1. B
ox Whisker plot showing
comparative antimicrobial
activity of pediocins.
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FIGURE 3. Images showing effect of pediocins on human spermatozoa a) normal spermatozoa; curved sperm tails; c) coiled sperm tails;
d) and e) sperm aggregation and immobilization.
FIGURE 4. B
ox Whisker plot showing
differences in spermicidal
activities of native and recpediocin.
Discussion
Probiotic LAB strains are greatly appreciated for fermentation
and preservation of dairy and meat products [47]. Probiotic
LAB synthesize a variety of antagonistic factors such as antimicrobial peptides or bacteriocins, lactic acid, diacetyl, hydrogen peroxide, fungicidal agents etc. that will force the opportunistic pathogens away from the ecological niche conquered
back by the healthy probiotic microflora [48-50]. AMPs or
bacteriocins from GRAS bacterial species have aroused a
great deal of industrial interest as they are extremely appealing alternatives to conventional pharmaceutical treatments.
Since these bacteriocins are prone to proteolytic digestion
in the gastrointestinal tract and therefore seem to lack any
potential antigenicity/toxicity in animals [51-52]. AMPs kill
their target cells by inducing drastic changes in membrane
permeability and disrupting proton motive force [53-54].
Many AMPs exhibit structural similarities with members of
eukaryotic channel forming peptides [53]. They may hinder
biosynthesis of DNA, peptidoglycan, proteins and RNA as a
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result of PMF depletion, interfere with the activities of essential enzymes, degrade vital macromolecules such as DNA,
RNA and cause cell lysis [53, 55-56].
Much of the earlier research has been carried out to investigate the biopreservative potential of antimicrobial peptides
synthesized by LAB, but very little has been reported on demonstrating their efficacy in treatment of human diseases such
as BV. In light of above as well as improvement in understanding and knowledge of the complex microbial interactions,
an increase tendency to use live probiotic LAB is urgently
desired for formulating therapeutic health care substances
aimed at prophylactic and control of human gastrointestinal
and urogenital diseases. Colonization of the vaginal ecosystem by health promoting probiotic LAB particularly prevents
infection by competition for available nutrients and mannose
sugar and by interfering binding of pathogens to the cell surface receptors [57-58]. An acidic pH of vaginal secretion aids
in keeping microbial infections away. However, low vaginal
pH alone is not sufficient to inhibit vaginal pathogens and to
prevent BV [48-49]. Thus, bacteriocin based therapeutics are
urgently desired to cure BV as well as to overcome side effects of antibiotic therapy such as diarrhea, poor compliance
and recurrence of vaginal infections.
Both native and rec-pediocin CP2 are also extremely attractive
therapeutic agents aimed at control of gastro-intestinal infections caused by food spoilage bacteria and general opportunistic human pathogens such as Bacteroides, Clostridium,
Coliforms, H. pylori, Listeria, Propionibacterium, Pseudomo
nas, Salmonella, Staphylococcus and Streptococcus sps. There
is increasing body of evidence that indicate potential of probiotic strains in cure and prevention of gut infections such as rotavirus diarrhea [59], travellers diarrhea [60], antibiotic associated diarrhea [61], necrotizing enterocolitis [62-63], inflammatory bowel disease [64-65], H. pylori associated infection
[66-67] and atopic diseases [68]. Growing scientific evidences
have proven efficacy of probiotics in maintaining restoring
and gut homeostasis that have been impressively reviewed
recently by Thirabunyanon in his article on biotherapy for
and protection against gastrointestinal pathogenic infections
via action of probiotic bacteria [37]. They are supporting
our investigation as Pediococci possess desirable antimicrobial properties for formulating probiotic dietary supplements,
yogurts, drinks and capsules [1-4].
Most of the published work regarding in vivo efficacy of
bacteriocins has focused on the oral delivery of probiotic
LAB to the gastrointestinal tract, where they will establish
themselves and secrete antagonistic bacteriocins. The use
of live probiotic bacteria may have prophylactic applications,
but the use of purified AMPs appears to be more attractive
for countering an established bacterial infection [69]. Experi Under License of Creative Commons Attribution 3.0 License
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Conclusions
Class IIa pediocins are antagonistic to many important human
pathogens. These bacteriocins have the ability to target a relatively wide range of pathogenic bacteria which is an important
advantage, especially to replace antibiotics with much safer
therapeutics and to combat the ever growing problem of antibiotic resistance, when compared to other antibiotics. They
have the potential to perform a very specific role like treatment of bacterial vaginosis, gut infections and peptic ulcers.
Preliminary experiments with rec-pediocin CP2 have proved it
to be effective at fighting Bacteroides, Candida, Escherichia,
Enterococcus, Helicobacter, Gardnerella, Klebsiella, Listeria,
Neisseria, Propionibacterium, Staphylococci, Streptococci and
Vibrio infections compared to native pediocin CP2. Attempt
at engineering pediocin CP2 of P. acidilactici MTCC 5101 to
improve its potency and stability has been highly encouraging.
Study also established pediocins as potent spermicidal agents
with very impressive market value in the form of personal care
products. The application of class IIa pediocins as therapeutics
is a rapidly developing area, and there is still much to investi-
10
Acknowledgement
Authors acknowledge the UGC, New Delhi, India, for providing financial assistance in the form of Rajiv Gandhi National
Fellowship to Mr. Balvir Kumar.
References
1. Kaur B and Balgir PP (2004) Purification, characterization and
antimicrobial range of bacteriocin obtained from an isolate of
Pediococcus spp. J Punjab Acad Sci 1(2): 139-144.
2. Kaur B and Balgir PP (2007) Pediocin CP2 gene localization to plasmid
pCP289 of Pediococcus acidilactici MTCC 5101. Internet J Microbiol
3(2).
3. Kaur B and Balgir PP (2008) Biopreservative Potential of a broad-range
pediocin CP2 obtained from Pediococcus acidilactici MTCC 5101.
Asian J Microbiol Biotechnol Environ Sci 10(2): 439-444.
4. Balgir PP, Bhatia P, Kaur B (2010) Sequence analysis and homology
modeling to assess structure-function relationship of pediocin CP2 of
Pediococcus acidilactici MTCC 5101. Ind J Biotechnol. 9: 431-434.
5. Kaur B, Kumar B, Balgir PP (2012) Cloning and expression rec-pediocin
CP2 in Escherichia coli using OmpA and TAP gene fusion approach,
communicated.
6. Tominaga T and Hatakeyama Y (2007) Development of innovative
pediocin PA-1 by DNA shuffling among class IIa bacteriocins. Appl
Environ Microbiol 73(16): 5292-5299.
7. Amsel R, Totten PA, Spiegel CA, Chen CS, Eschenbach D, et al.
(1983) Nonspecific vaginitis: diagnostic criteria and microbial and
epidemiologic associations. Am JMed 74: 14-22.
8. Cherpes TL, Meyne LA, Krohn MA, Hiller SL (2003) Risk factors for
infection with herpes simplex virus type 2: role of smoking, douching,
uncircumcised males, and vaginal flora. Sex Transm Dis 30: 405-410.
9. Falagas ME, Betsi GI, Athanasiou S (2007) Probiotics for the treatment
of women with bacterial vaginosis. Clinical Microbiol Infection 13(7):
657-664.
iMedPub Journals
2012
ARCHIVES OF CLINICAL MICROBIOLOGY
32. Hay P (2000) Recurrent bacterial vaginosis. Curr Infect Dis Rep 2(6):
506-512.
33. Tosun I, Karaoglu SA, Ciftci H, Buruk CK, Aydin F, et al. (2007)
Biotypes and antibiotic resistance patterns of Gardnerella vaginalis
strains isolated from healthy women and women with bacterial
vaginosis. Mikrobiyol Bul 41(1): 21-27.
34. Nagaraja P (2008) Antibiotic resistance of Gardrenalla vaginalis in
recurrent bacterial vaginosis. Ind J Med Microbiol 26(2): 155-157.
35. Reddy KV, Aranha C, Gupta SM, Yedery RD (2004) Evaluation of
antimicrobial peptide nisin as a safe vaginal contraceptive agent in
rabbits: in vitro and in vivo studies. Reproduction 128: 117-126.
36. Sutyak KE, Anderson RA, Dover SE, Feathergill KA, Aroutcheva AA, et
al. (2008) Spermicidal activity of the safe natural antimicrobial peptide
subtilosin. Infect Dis Obstet Gynecol 2008: 540758.
37. Thirabunyanon M (2011) Biotherapy for and protection against
gastrointestinal pathogenic infections via action of probiotic bacteria.
Maejo Int J Sci Technol 5(01): 108-128.
38. Hooper LV, Midtwedt T, Gordon JI (2002) How host microbial
interactions shape the nutrient environment of the mammalian
intestine. Annu Rev Nutr 22: 283-307.
39. Guarner F and Malagelada JR (2003) Gut flora in health and disease.
Lancet 361: 512-519.
4 0. Thirabunyanon M, Boonprasom P, Niamsup P (2009) Probiotic
potential of lactic acid bacteria isolated from fermented dairy milks on
antiproliferation of colon cancer cells. Biotechnol Lett 31: 571-576.
41. Shmuely H, Passaro D, Figer A, Niv Y, Pitlik S, et al. (2001) Relationship
between Helicobacter pylori CagA status and colorectal cancer. Am J
Gastroenterol 96: 3406-3410.
42. Travaglione S, Fabbri A, Fiorentini C (2008) The Rho-activating
CNF1 toxin from pathogenic E. coli: A risk factor for human cancer
development? Infect Agent Cancer 3: 4.
43. Yang R, Johnson MC, Ray B (1992) Novel method to extract large
amounts of bacteriocin from lactic acid bacteria. Appl Environ
Microbiol 58: 3355-3359.
4 4. Cintas LM, Rodriguez JM, Fernandez MF, Sletten K, Nes IF, et al. (1995)
Isolation and characterization of pediocin L50, a new bacteriocin from
Pediococcus acidilactici with a broad inhibitory range. Appl Environ
Microbiol 61: 2643-2648.
45. Pucci MJ, Vedamuthu ER, Kunka BS, Vandenbergh PA (1988) Inhibition
of Listeria monocytogenes by using bacteriocin PA-1 produced by
Pediococcus acidilactici PAC1.0. Appl Environ Microbiol 54: 23492353.
4 6. Bakst MR and Sexton TJ (1979) Fertilizing capacity and ultrastructure
of fowl and turkey spermatozoa before and after freezing. J Reprod
Fertil 55: 1-7.
47. Gillor O, Etzion A, Riley MA (2008) The dual role of bacteriocins as
anti- and probiotics. Appl Microbiol Biotechnol 81: 591-606.
4 8. Redondo-Lopez V, Cook RL, Sobel JD (1990) Emerging role
oflactobacilliin the control and maintenance of the vaginal bacterial
microflora. Rev Infect Dis 12: 856-872.
49. Tomas MSJ, Ocana VS, Wiese B, Nader-Macias ME (2003) Growth and
lactic acid production by vaginal Lactobacillus acidophilus CRL1259,
and inhibition of uropathogenic Escherichia coli. J Med Microbiol 2:
1117-1124.
50. Zhou X, Brown CJ, Abdo Z, Davis CC, Hansmann MA, et al. (2007)
Differences in the composition of vaginal microbial communities found
in healthy caucasian and black women. ISME J 1: 121-133.
51. Bhunia AK, Johnson MC, Ray B, Belden EL (1990) Antigenic property
of pediocin AcH produced by Pediococcus acidilactici H. J Appl
Bacteriol 69: 211-215.
52. Kheadr E, Zihler A, Dabour N, Lacroix C, Blay GL, et al. (2010) Study
of the physicochemical and biological stability of pediocin PA-1 in the
upper gastrointestinal tract conditions using a dynamic in vitro model.
J Appl Microbiol 109(1): 54-64.
53. Montville TJ and Brune MEC (1994) Evidence that dissipation of proton
motive force is a common mechanism of action for bacteriocins and
other antimicrobial proteins. Inter J Food Microbiol 24: 53-74.
11
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