ANRV322-MI61-23 ARI 6 August 2007 18:7

Lantibiotics: Peptides
of Diverse Structure
and Function
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org

Joanne M. Willey1 and Wilfred A. van der Donk2

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1
Department of Biology, Hofstra University, Hempstead, New York 11549;
email: biojmw@hofstra.edu
2
Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana,
Illinois 61801; email: vddonk@uiuc.edu

Annu. Rev. Microbiol. 2007. 61:477–501 Key Words

First published online as a Review in Advance on
June 18, 2007
The Annual Review of Microbiology is online at
micro.annualreviews.org
This article’s doi:
10.1146/annurev.micro.61.080706.093501
Copyright  c 2007 by Annual Reviews.
All rights reserved
0066-4227/07/1013-0477$20.00
bacteriocin, peptide antibiotic, peptide engineering,
posttranslational modification
Abstract
The current need for antibiotics with novel target molecules has co-
incided with advances in technical approaches for the structural and
functional analysis of the lantibiotics, which are ribosomally synthe-
sized peptides produced by gram-positive bacteria. These peptides
have antibiotic or morphogenetic activity and are structurally defined
by the presence of unusual amino acids introduced by posttransla-
tional modification. Lantibiotics are complex polycyclic molecules
formed by the dehydration of select Ser and Thr residues and the
intramolecular addition of Cys thiols to the resulting unsaturated
amino acids to form lanthionine and methyllanthionine bridges, re-
spectively. Importantly, the structural and functional diversity of the
lantibiotics is much broader than previously imagined. Here we dis-
cuss this growing collection of molecules and introduce some re-
cently discovered peptides, review advances in enzymology and pro-
tein engineering, and discuss the regulatory networks that govern
the synthesis of the lantibiotics by the producing organisms.

477
 ANRV322-MI61-23 ARI 6 August 2007 18:7

protein structures. In this review, we discuss

Contents the lantibiotics, with an emphasis on recent
findings. The reader is directed to a number
INTRODUCTION . . . . . . . . . . . . . . . . . 478
of other reviews that offer a more historical
AN OVERVIEW OF
perspective (12, 21, 94, 117).
LANTIBIOTIC
STRUCTURES . . . . . . . . . . . . . . . . . 478
BIOLOGICAL ACTIVITIES AND AN OVERVIEW OF
APPLICATIONS . . . . . . . . . . . . . . . . 479 LANTIBIOTIC STRUCTURES
CLASSIFICATION SCHEMES . . . . . 481
Lantibiotics are small (19–38 amino acids)
SELF-IMMUNITY . . . . . . . . . . . . . . . . . 481
peptides that undergo extensive posttrans-
LANTIBIOTIC STRUCTURE
lational modification. The least modified
AND FUNCTION . . . . . . . . . . . . . . 484
peptide to date is lactocin S (24%), and
Class I Lantibiotics: Nisin,
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org

the recently described compound 107891

Subtilin, and Related Peptides . . 484
is the most heavily modified (58%) (65).
Class II Lantibiotics . . . . . . . . . . . . . . 485
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The posttranslational modifications common

Class III Lantibiotics: SapB and
SapT . . . . . . . . . . . . . . . . . . . . . . . . .
Posttranslational Modification
Enzymes and Engineering
Studies . . . . . . . . . . . . . . . . . . . . . . . .
LANTIBIOTIC GENETICS . . . . . . .
Biosynthetic Gene Clusters . . . . . . .
Regulation of Lantibiotic Synthesis
CONCLUSIONS AND
OUTLOOK . . . . . . . . . . . . . . . . . . . . .
INTRODUCTION
Lantibiotic: The lantibiotic nisin (Figure 1) produced by
gene-encoded, Lactococcus lactis was first discovered in 1927
posttranslationally
(92). This peptide has become the best under-
modified peptide
containing one or stood of over 50 lantibiotics produced by both
more Lan and/or low and high G+C gram-positive bacteria.
MeLan residues Lantibiotics are defined by posttranslational
Propeptide: modifications that introduce the thioether
C-terminal segment amino acids lanthionine and methyllanthio-
of prepropeptide that nine; in fact the term lantibiotic is derived
is posttranslationally
from lanthionine-containing antibiotics (96).
modified to
ultimately become Although the lantibiotics were once thought
the mature to be a relatively small group with limited bio-
lantibiotic logical activity, a larger collection of peptides
Dha: 2,3- is emerging with a widening array of struc-
didehydroalanine tures and functions. These molecules are in-
Dhb: (Z )-2,3- triguing because of their unique biochemistry,
didehydrobutyrine genetic regulation, range of biological func-
tions, and potential for engineering unique
to all lantibiotics involve the dehydration
487
of Ser and Thr residues in the propep-
tide to yield 2,3-didehydroalanine (Dha) and
(Z )-2,3-didehydrobutyrine (Dhb), respec-
487
tively (Figure 1a). This is followed by the
488
stereospecific intramolecular addition of a
488
Cys residue onto Dha or Dhb to form a lan-
489
thionine (Lan) or methyllanthionine (MeLan)
bridge, respectively. A representative example
492
is shown in Figure 1b for nisin.
The modified peptide is then exported
and a leader sequence (23- to 59-amino-acid
residues) is proteolytically removed, although
not always in that order. Lantibiotic leader se-
quences are conserved among groups of lan-
tibiotics (12) but bear no sequence homology
to the Sec-dependent or twin-arginine (TAT)
exported proteins, with the exception of the
leader peptide of cinnamycin (123). In cases
in which the function of the leader has been
specifically addressed, it is required for secre-
tion (15, 58, 90, 110, 115) and ensures com-
plete maturation (14, 29, 66, 130). For those
lantibiotics that are proteolytically processed
extracellularly, the leader may prevent activa-
tion of the lantibiotic until it is safely outside
the cell (114), as lantibiotics with their leader
sequence attached have no antimicrobial ac-
tivity (62, 66, 114, 130).
At present, no fewer than 15 differ-
ent posttranslational modifications have been
documented in lantibiotics. This structural

478 Willey · van der Donk

 ANRV322-MI61-23 ARI 6 August 2007 18:7

a H O H
O b NisA
N N Leader-ITSISLCTPGCKTGALMGCNMKTATCHCSIHVSK
Xn
R OH SH - 8 H2O NisB
(Dehydratase)
Dehydration

O H O
H Leader-IDhbDhaIDhaLCDhbPGCKDhbGALMGCNMKDhbADhbCHCSIHVDhaK
N N
Xn
R S NisC
H (Cyclase)
R=H Dha
R = CH3 Dhb
A C E
S S S
Cyclization
Leader-IDhbAlaIDhaLAlaAbuPGAlaKAbuGALMGAlaNMKAbuAAbuAlaHAlaSIHVDhaK
Xn S S
B D
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org

Proteolysis NisP
O S NH (Protease)
D L
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HN R O
O Ala Leu
HN Met
R=H (2S,6R)-lanthionine
R = Me (2S,3S,6R)-3-methyllanthionine Gly
Ile Leu O Gly
H
O NH N O
S NH NH S
Lys His
S NH Lys
HN O S O Me O NH
O Me NH
O O Asn Met
Me NH Ala O O
HN Pro Gly N
H
Ile Me S Ser
Me Ile
Nisin A Val His
Lys
N
O H

Figure 1
(a) Installation of lanthionine (Lan) or methyllanthionine (MeLan) residues into lantibiotic prepeptides.
(b) The posttranslational maturation process of nisin. For Lan and MeLan structures, the segments
derived from Ser/Thr are in red and those derived from Cys are in blue.

diversity releases the peptides from the con- isoleucine (76). The antibiotic 107891 con-
straints imposed by the use of only 20 amino tains two previously unknown modifications:
acids (Figure 2). For instance C-terminal a chlorinated Trp and a mono- or dihydroxy-
Lan: lanthionine
Cys residues may form S-aminovinyl-d- lated Pro (65).
MeLan:
cysteine (AviCys) or S-aminovinyl-3-methyl-
3-methyllanthionine
d-cysteine (AviMeCys) structures, and Dha
and Dhb residues that are N-terminally ex-
BIOLOGICAL ACTIVITIES AviCys: S-[(Z)-2-

posed after leader processing spontaneously

AND APPLICATIONS aminovinyl]-d-
cysteine
hydrolyze to yield 2-oxopropionyl (OPr) and At a time when there is an urgent need
AviMeCys: S-[(Z)-
2-oxobutyryl (OBu) groups (50). OPr may for new antibiotics, lantibiotics demon- 2-aminovinyl]-d-3-
be further modified by reduction to a 2- strate promising chemotherapeutic poten- methylcysteine
hydroxypropionyl (Hop) residue (30, 111). tial. Many lantibiotics are bacteriocidal OPr:
Moreover, l-Ser can be converted to d-Ala against a variety of gram-positive bacteria 2-oxopropionyl
with Dha as an intermediate (20). Cinnamycin at nanomolar levels and some are active OBu: 2-oxobutyryl
and the closely related duramycins have a lysi- against methicillin-resistant Staphylococcus au-
Hop:
noalanine bridge, as well as a hydroxylated reus (MRSA), vancomycin-resistant entero- 2-hydroxypropionyl
Asp (48, 135), and cypemycin has an allo- cocci, and oxacillin-resistant gram-positives

www.annualreviews.org • Lantibiotic Function and Diversity 479

 ANRV322-MI61-23 ARI 6 August 2007 18:7

O NH O Me NH O
O H
(D) (L) (D) (L) H N
N
HN S O HN S O
H3C
meso-lanthionine (2S,3S,6R)-3-methyllanthionine 2,3-Didehydroalanine (Z)-2,3-didehydrobutyrine

Ala Ala Abu Ala Dha Dhb

S S

O
O O
H
N CH3
O NH O H
(L)
(CH2)3 (L)
(D)
HO
O 2-Oxopropionyl OPr
HN N O HN S NH
H OH
O
(2S,9S)-lysinoalanine AviCys erythro-3-hydroxy-
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org

H3C
L-aspartic acid
O
Ala
Ala Lys S Asp OH 2-Oxobutyryl OBu
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N
H NHR
O
O HO
O Me H O
H
H2N (D) N CH3
HN S NH
2-Hydroxypropionyl Hop
NH
Figure 2
allo-isoleucine AviMeCys ClTrp O
Posttranslational
N
modifications that O Abu Cl
H HPro
have been reported N S
D-Ala HO OH
to date for the NHR
lantibiotic family. Dihydroxyproline

(22). To date only nisin has been used com- of phospholipase A2 (69). Duramycin in-
mercially, with a 40-year history as a preser- creases chloride secretion in lung epithelium
vative in food products. There is interest in (16) and has been in clinical trials to evalu-
expanding its use to food packaging, as well as ate its efficacy in clearing mucus secretions
introducing it to clinical applications in both from the lungs associated with cystic fibro-
human and animal health products (22, 117). sis and other airway diseases (77). The ente-
Some interesting clinical applications have rococcal lantibiotic cytolysin not only targets
been proposed and/or are in clinical trials. other gram-positives, but functions as a vir-
The lantibiotic mutacin 1140 is produced ulence factor, lysing erythrocytes and poly-
by Streptococcus mutans, the major causative morphonuclear leukocytes (24). Perhaps the
agent of dental caries. The compound is effec- most novel activity is found in the peptides
tive against many strains of the same species, SapB and SapT, produced by the filamen-
prompting interest in this peptide as an agent tous soil microbes Streptomyces coelicolor and
to prevent dental caries (36, 120). Along sim- S. tendae, respectively (54, 55). Streptomyces
ilar lines, lozenges seeded with a salivaricin spp. undergo a complex, fungal-like life cycle
A–producing strain of S. salivarius have been that involves the differentiation of substrate-
introduced as a probiotic to combat S. pyogenes associated hyphae into upwardly growing
in the oral cavity and prevent halitosis (9, 122). aerial hyphae that then form reproductive
Not all lantibiotics function strictly as an- spores. SapB (54) and SapT (55) are both hy-
timicrobial agents, however. Cinnamycin and drophobic and surface active; they are thought
the related duramycins are potent inhibitors to function as biosurfactants, releasing the

480 Willey · van der Donk

 ANRV322-MI61-23 ARI 6 August 2007 18:7
surface tension at the colony-air interface
to facilitate the emergence of nascent aerial
hyphae (105).
Thus, while lantibiotics were initially dis-
covered, further isolated, and explored as
bacteriocins, the variety of lantibiotic activi-
ties now realized suggests that their functional
diversity may be underestimated. By screen-
ing only for antibiotic activity, lantibiotics that
demonstrate other, unexpected biological ac-
tivities may have been missed.
CLASSIFICATION SCHEMES
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In this review we adopt a classification scheme
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that places all lantibiotics in one of three
classes (Class I, II, or III) on the basis of
the pathway by which maturation of the pep-
tide occurs and the presence or absence of
antibiotic activity. This designation incorpo-
rates differences in leader peptide sequence,
biosynthetic operon structure, peptide func-
tion, and to a lesser extent, the structure of the
mature lantibiotic. This approach is based on
a scheme first suggested by Pag & Sahl (84)
and is simple, is inclusive, and may be suffi-
ciently flexible to accommodate lanthionine-
containing peptides that have not yet been
discovered.
Class I lantibiotic prepeptides are mod-
ified by two distinct enzymes: the gener-
ically termed LanB enzyme, which dehy-
drates Thr and Ser residues, and LanC, which
mediates cyclization. The peptides are ex-
ported by LanT, a dedicated ABC transporter.
Class I leader peptides are cleaved by LanP,
subtilisin-like serine proteases. These lantibi-
otics are more linear than class II (Figure 3).
Class II lantibiotics are modified by the
LanM-type modification enzymes, which are
large (900–1000 amino acids) proteins that ex-
hibit both dehydratase and cyclase activities.
Whereas LanM enzymes bear no homology
to LanB proteins, they exhibit low sequence
identity to the LanC enzymes (97), includ-
ing three zinc binding ligands (83). Class II
lantibiotic secretion and leader processing is
also performed by a single, multifunctional
protein with a conserved N-terminal cys-
teine protease domain (84). Despite the func-
tional difference between class I and class II
Bacteriocin:
exporters, they share the designation LanT. bacterial proteins
Unique within the class II molecules are the that demonstrate
two-component lantibiotics, in which two antibiotic activity
peptides assemble to form the mature antibi- against other strains
of the same species
otic. Each peptide is encoded by its own struc-
or closely related
tural gene and modified by separate LanM species
enzymes (71, 73). However, a single LanT
protein removes the leader peptide and se-
cretes both products.
We propose that a new, third class be es-
tablished for lanthionine-containing peptides
that lack significant antibiotic activity but in-
stead perform another function for the pro-
ducing cell. To date three such peptides have
been described: S. coelicolor secretes SapB (54,
55, 128), AmfS is produced by S. griseus (109),
and SapT is the product of S. tendae (55,
128). Of these morphogenetic peptides, only
the structure of SapT has been confirmed by
NMR spectroscopy; the structure of SapB is
inferred from its amino acid sequence and
mass spectrometry (54). AmfS has been puri-
fied, but its structure, like that of three other
SapB orthologues (two produced by S. scabies
and one by S. avermitilis), has not been rigor-
ously explored (128).
Apart from their unusual function, other
factors further distinguish SapB and its known
orthologues from antimicrobial lantibiotics.
Although the putative modification enzymes
for SapB and AmfS (RamC and AmfT, respec-
tively) bear homology to the C-terminal cy-
clization domain of LanM enzymes, they lack
the zinc ligands that are important for cycliza-
tion by LanM. Thus, the mechanism of mat-
uration of these class III lantibiotics may be
distinct.
SELF-IMMUNITY
Antibiotic synthesis requires that producer or-
ganisms have a mechanism conferring resis-
tance to their product. In the case of lan-
tibiotics, immunity can be provided by a
specific immunity protein (LanI) and/or by
www.annualreviews.org • Lantibiotic Function and Diversity 481
 ANRV322-MI61-23 ARI 6 August 2007 18:7

specialized ABC transporter proteins encoded for self-immunity exists for the Bacillus subtilis
by the lanFEG genes. The mechanism of Pep5 product subtilin (102), but despite the struc-
self-immunity is perhaps one of the simplest; tural similarity between the two lantibiotics,
a single PepI peptide appears to bind at the NisI cannot protect L. lactis from subtilin
membrane–cell wall interface, thereby mask- (103). Another variation of the LanI/LanFEG
ing the Pep5 target molecule (38). In many theme is represented by NukH, which helps
cases, however, LanI cannot confer full resis- confer immunity to nukacin ISK-1. NukH
tance and it functions cooperatively with the differs from NisI-like proteins in that it is
ABC transporters. NisI, which has a lipopro- smaller and has three transmembrane do-
tein signal sequence attaching it to the outer mains. NukH functions synergistically with
face of the membrane, intercepts nisin to re- the NukFEG transporter to provide self-
duce its local concentration in the immedi- protection as well as immunity to the struc-
ate environment (103). A similar mechanism turally similar lantibiotic lacticin 481 (2).
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org

Class I
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Nisin A Ala Leu

Gly Met S Ser Ile His Val Dha Lys OH
E Ala
Abu C Abu
Gly Ala
Dha Ala His
H Ile Ile Leu Lys Ala D
S S Abu S
Dhb Ala A Ala Abu B Ala Asn
Lys
Pro Gly Met
S

Ala Leu
Figure 3 Subtilin Gly Gln S Lys Ile Dha Lys OH
Ala
Abu
Representative Dha
Abu Dhb Leu Ala Asn
examples of the H Trp Glu Leu
Val S Ala
S Abu
three classes of Lys Ala Ala S
Abu Ala Phe
lantibiotics. The Gln
S Pro Gly Leu
same color-coding
and shorthand
notation is used as Epidermin Asn Ala S
Phe
defined in H Lys Ile Phe Tyr
Ile S
Figure 1. In most Ala
NH
Ala Ala Ala Abu Ala Lys Ala Ala
cases the Ser/Thr S
S
is located Pro Gly Dhb Gly

N-terminal of the
cysteine that forms
107891
a Lan/MeLan with Dha
ClTrp Leu
it. Exceptions are S Thr Ala
S
Ala
S
Ala
found in H Val Dhb Ala Ala Abu Ala Phe
cinnamycin S HPro Asn
Pro Gly Ala NH
(Cys1 + Thr18, Gly Ala
S
Cys5 + Thr11) Gly Gly

(40, 135),
mersacidin Val
Lys
Gln
(Cys1 + Thr2) Pep5 Lys OH
Ala Ala Ala Gln Lys Dhb Leu Phe Abu Val
(94), and the S S Ala
α-peptide of Arg Leu Arg Ala
Gly
S
lacticin 3147 OBu Ala Gly Pro Ala Ile Lys Ala Dhb Ala
Asn
(Cys1 + Thr2) Lys
(70). Gly Lys

482 Willey · van der Donk

 ANRV322-MI61-23 ARI 6 August 2007 18:7

Class II

Cinnamycin Lacticin 481 Mersacidin

S
OH
11 Ser Gly Dhb Gly
Phe Abu Phe HO Phe H Gly
Gly Asp Ala Gly
Val Pro Gly
Asn B Val Ala Ser Ala Val Ala Ile
Ala14 Ala Pro Gly
S S C Gly Gly Ile His C B S
S Ile S Glu
Abu S Phe Abu Leu Val HN
A His Abu A Glu B
Ala1 4 Phe Lys
Gln Ala Ala Trp Gln C Dha D
Lys D Arg Ala5 H Ala Phe Abu Ala
Ala6 S Abu S
Asn Abu
HO Asn Met S Leu
N
H

Lactocin S Sublancin
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Trp Leu
Ala
Leu Leu
Val Ala His His Gly Gly
Dha Ala Gln Ala
Abu Gln Lys
Pro Lys Ala Cys
Pro D-Ala Thr Ala Ala OH Ile Cys
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Leu Val Gly

Gly Ala S S
Dhb Val Ala Asp Gly S
Leu S S Leu
OPr Ala Ala Ala S
D-Ala Ala Cys
H Glu Ala Gly
S Cys
Val Val Phe Tyr Gly Phe Arg
Met Tyr Ala Gln
D-Ala Leu Lys Gly Val Asn Gln OH H
Gly Ala
Tyr Arg

Lacticin 3147 A1 (Ltn α) Lacticin 3147 A2 (Ltn β)

Asn
Gly Asn
Dhb Asn Pro Ala
Trp Gly Ile Asn S
Dhb Dhb Thr
Ala Thr OH
S A B Ala Met Ala C Ala
Phe Tyr Ala Ala D-Ala A Abu
Ala S Glu Ala Arg Ala
Ala Ile Ile S S
Asp Trp Pro Ala
D-Ala
H C His D Trp Leu Tyr Pro B
Leu Ala Ala OBu Dhb Abu Thr Lys
Abu Ala
Abu Ala D-Ala
S Leu
S Lys OH

Haloduracin α Haloduracin β
Gly Asn Lys
Leu
Arg Gly Ala Dhb
Val Ala
A Ala Met Ala Val S
Ala Gly OH
H2N O Val Ala Leu D Ala
S Glu Pro Pro B Abu
NH Tyr
A S S Ala Dha Gln
O S S B Val C Ser Trp S Ala
Ala Ala C
Abu Pro
Ala Abu Ala Dhb
Ile S Leu Abu Thr Lys
Trp Tyr Asn S Asn OH Abu H

Class III
SapB SapT
S S
H Thr Gly Ala Ala Gly Ala Ala Ala Asn OH
S S S S
Arg Leu Ser Thr H Tyr Abu Ala Ala Ala Abu Ala Ala Abu Ala Gly OH
Gln Gly Gly Leu
Ala Leu Leu Thr Ile Ile Val Ile
Val Val
Dha Leu Dha Ile

Figure 3
(Continued )

www.annualreviews.org • Lantibiotic Function and Diversity 483

 ANRV322-MI61-23 ARI 6 August 2007 18:7

LANTIBIOTIC STRUCTURE a flexible hinge region (112). The MeLan and

AND FUNCTION Lan bridging patterns found in subtilin and
ericin S produced by B. subtilis ATCC6633
Prepropeptide: Our understanding of the various modes of
precursor peptide for and B. subtilis A1/3, respectively (99), are iden-
action of lantibiotics has greatly increased
posttranslational tical to that of nisin. B. subtilis A1/3 also pro-
in recent years, as advances in peptide engi-
modification. Also duces ericin A, in which the three N-terminal
neering have facilitated the investigation of
called prepeptide bridges are conserved, but differs from sub-
structure-activity relationships. A short sum-
Lipid II: tilin by 13 residues and the position of the
mary of these developments is provided in the
undecaprenyl- two C-terminal MeLan residues. The gene
pyrophosphoryl- following sections on select members of the
clusters encoding subtilin and ericin A and S
MurNAc- three classes of lantibiotics defined above.
(pentapeptide)- are strikingly similar, with the exception that
GlcNAc, B. subtilis A1/3 possesses two prepropeptide
membrane- Class I Lantibiotics: Nisin, Subtilin, structural genes. It has been speculated that
and Related Peptides
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org

associated this cluster underwent two duplications of the

peptidoglycan
L. lactis strains produce at least three struc- subtilin structural gene, spaS, to give rise to
monomer that is
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polymerized and tural variants of nisin. Nisin A is the proto-
cross-linked to type (Figure 1), nisin Z differs by one amino
generate the acid, and nisin Q differs at four positions;
bacterial cell wall all are 34 residues in length (27). Streptococ-
cus uberis produces an additional variant, nisin
U, which has 78% sequence identity to nisin
A and lacks the C-terminal three residues
(129). Nisin is a flexible molecule with two
amphiphilic domains consisting of three N-
terminal rings (labeled A, B, and C) and the
C-terminal D and E rings, which are joined by
the two ericin paralogues (99).
The pore-forming activity of nisin is
unique in that the cell wall precursor lipid II is
used as a docking molecule (6, 8) (Figure 4).
The N-terminal amides of the A and B rings
of nisin form a cage that binds the pyrophos-
phate moiety of lipid II (33, 43, 44, 127).
This binding motif is also found in other lan-
tibiotics that interact with lipid II, including
subtilin, epidermin, gallidermin, and mutacin
1140 (4, 34). Epidermin and gallidermin lack

Figure 4
The biosynthesis
of the bacterial cell
wall with the
binding sites for
the glycopeptide
antibiotic
vancomycin and
the lantibiotic nisin
indicated. By using
lipid II as a docking
molecule, nisin
inhibits
polymerization of
murein subunits in
addition to forming
pores in the plasma
membrane.

484 Willey · van der Donk

 ANRV322-MI61-23 ARI 6 August 2007 18:7
the C-terminal tail and are shorter (∼30 Å)
compared with nisin (50 Å). This difference
may explain why these two compounds do not
form pores in bacteria that are susceptible to
pore formation by nisin (4).
The C terminus of nisin inserts into the
membrane in a perpendicular orientation with
respect to the bilayer surface (44, 116), re-
sulting in stable pores 2–2.5 nm in diameter
(124) consisting of eight nisin and four lipid II
molecules (33). This stoichiometry differs
from the NMR structure of the 1:1 nisin–
lipid II complex, and it is possible that nisin-
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org
nisin interactions account for the difference.
Attaining a transmembrane structure requires
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the presence of the flexible hinge region; nisin
Z peptides mutated in this region are unable
to form pores but retain the ability to bind
lipid II (127, 133). Breukink and coworkers
(34) recently demonstrated that nisin and mu-
tacin 1140 disrupt the functional localization
of lipid II. This cell wall precursor is nor-
mally found at sites of nascent cell wall biosyn-
thesis such as the septum and along helical
bands encircling the cylindrical axis of rod-
shaped bacteria (25, 106, 113). Nisin-treated
cells sequester lipid II in patches on the bac-
terial membrane, removing it from the site of
cell wall biosynthesis. The extreme sensitiv-
ity observed in susceptible bacteria and the
absence of nisin-resistant mutants reflect the
dual modes of nisin activity: pore formation
coupled with the inhibition of cell wall biosyn-
thesis (33, 34).
Class II Lantibiotics
The class II lantibiotics are structurally di-
verse but are similar in that they use a single
enzyme to catalyze dehydration and cycliza-
tion. It currently represents the largest of the
three classes of lantibiotics.
Lacticin 481 and its relatives. Lacticin 481
is one of about 20 related lantibiotics (29). All
are hydrophobic with no net charge at neu-
tral pH. In all cases, the N terminus is linear
and three overlapping thioether bridges ren-
der the C terminus globular (Figure 3). Site-
directed mutagenesis of mutacin II shows that
its hinge is essential for antibiotic activity and
while all three thioether linkages are impor-
tant, if not required, for full activity (14), sub-
stitution of the MeLan bridge with Lan or
vice versa has no effect on activity in mutacin
II and lacticin 481 (11, 14, 29).
Lacticin 481 and its relatives are believed
to form pores in target microbes (29), al-
though the means by which this is accom-
plished has not been fully explored. The A
ring in this subfamily shows similarity with the
C ring of mersacidin, which is critical for its
activity (104). The observation that microbes
producing these lantibiotics have similar im-
munity proteins that prevent and/or remove
the peptide from the membrane further sug-
gests that the membrane is targeted (29).
Mersacidin. The discovery that mersacidin
is active against methicillin-resistant S. au-
reus in a murine model (57) has prompted ef-
forts to determine the key structural compo-
nents of this peptide. Unlike nisin and other
cationic peptides, mersacidin does not form
pores in plasma membranes. Instead, it ap-
pears to inhibit the transglycosylation step in
peptidoglycan biosynthesis (7). Although the
molecular details remain to be resolved, its
conformation changes in the presence of lipid
II (42), and the C ring (particularly the Glu
therein) is believed to be essential for recog-
nition (41, 104). In fact, this ring is conserved
in the lacticin 481 subgroup of lantibiotics,
the two-component lantibiotics (108), and in
plantaricin C (125).
Cinnamycin and the duramycins. Cin-
namycin and three duramycin variants are
streptomycete products (123) and are remark-
able for the presence of unusual modifications
including a lysinoalanine ring (Figures 2 and
3). They bind to phosphatidylethanolamine
(PE) with a 1:1 stoichiometry (119), inducing
transbilayer lipid movement (68).
www.annualreviews.org • Lantibiotic Function and Diversity 485
 ANRV322-MI61-23 ARI 6 August 2007 18:7

Two-component lantibiotics. Lantibiotics presumed to be important in lipid II bind-

that consist of two modified peptides in-
clude plantaricin W, staphylococcin C55, cy-
tolysin L, Smb, BHT-A, haloduracin, and
lacticin 3147. Both the mature peptides
and prepeptides are often called LanA1 and
LanA2; here we differentiate the two by us-
ing LanA1/LanA2 to refer to the unmodi-
fied prepeptides and Lanα/Lanβ to designate
the mature peptide subunits. In all cases ex-
cept cytolysin, the α-peptide resembles mer-
sacidin and the β-peptide is relatively long and
flexible (70) (Figure 5).
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org
ing. The N-terminal MeLan ring identified
in Halβ (73) appears to be conserved in all
β-peptides except lacticin 3147 and staphylo-
coccin C55. This ring is followed by a stretch
of hydrophobic amino acids, where Ser to d-
Ala conversions occur in LtnA2 of lacticin
3147. The B ring is also relatively conserved
among currently known two-component lan-
tibiotics, absent only in BHT and Smb. Fi-
nally, the two most C-terminal Lan/MeLan
rings are conserved in all β-peptides except
cytolysin.

To date only the structure of lacticin 3147 Each peptide of the two-component
has been fully elucidated by NMR spec- lantibiotics displays weak or no antimicrobial
by Pennsylvania State University on 05/22/12. For personal use only.

troscopy (70), with the structure proposed
for haloduracin strongly supported by tan-
dem mass spectrometry (73) (Figure 3). This
structural information combined with se-
quence homology suggests that the three C-
terminal rings in all α-peptides have the same
topology, which in analogy to mersacidin is
activity, but synergistic interaction of the
molecules results in strong antibiotic activity.
A nisin-like model for lacticin 3147 has been
proposed (126), in which Ltnα is thought
to bind lipid II and recruit Ltnβ to form
pores in target cytoplasmic membranes
(78, 126). A recent study in which every

a
A B C
HalA1 MTNLLKEWKMPLERTHNNSNPAGDIFQELEDQDILAGVNGA ––CAWYNI–SCRLGNKGAYCTLTVECMPSCN 69
PlwAα MK––ISKIEAQAR––KDFFKKIDTNSNLLNVNGA –KCKWWNI–SCDLGNNGHVCTLSHECQVSCN 59
SacAα MKSSFLEKDIEEQ––VTWFEEVSEQEFDDDIFGA CSTNTFSL–SDYWGNKGNWCTATHECMSWCK 62
LtnA1 MN––––KNEIETQP–VTWLEEVSDQNFDEDVFGA CSTNTFSL–SDYWGNNGAWCTLTHECMAWCK 59
BhtA1 MK–EIQKAGLQEEL–SILMDDAN––NLEQLTAGI GTTVVNSTFSIVLGNKGYICTVTVECMRNCQ 61
SmbA1 MK–EIQKAGLQEEL–SILMDDAN––NLEQLTAGI GTTVVNSTFSIVLGNKGYICTVTVECMRNCSK 61

b A B C D
HalA2 MVNSKDLRNPEFRKAQGLQFVDEVNEKELSSLAGSGDVHAQ-TTWPC––ATVGV––––SVALC–PTTKCTSQC 65
PlwAβ MTKTSRRKNAIANYLEPVDEKSINESFGAGDPEAR–SGIPC–TIGAAVAA––SIAVC–PTTKCSKRCGKRKK 67
BhtA2 MKSNLLKINNVTEVEKDMVTLIKDEDMELAGG–––––––STPAC––AIGVVGI––TVAVTGISTACTSRCINK 62
SmbA2 MKSNLLKINNVTEMEKNMVTLIKDEDMLAGG–––––––STPAC––AIGVVGI––TVAVTGISTACTSRCINK 61
LtnA2 MKEKNMKKNDTIELQLGKYLEDD–MIELAEGDESHGG–––––––TTPATP–AISILSAYISTNTC–PTTKCTRAC 65
SacAβ MKNELGKFLEENELELGKFSESDMLEITDDEVYAAGTPLALLGGAAT––GVIGYI–––SNQTCP–TTACTRAC 67
CylS-A MLNKENQENYYSNKLELVGPSFEE––LSLEEMEAIQGSGDVQAE–TTPACFTIGLGVGALFSAKFC 63
CytL-A MENLSVVPSFEELSVEEMEAIQGSGDVQAE–TTPVC–AVAATAAA––SSAACGWVGGGIFTGVTVVVSLKHC 68

Figure 5
Sequence alignment of (a) the LanA1 and (b) the LanA2 prepeptides of haloduracin (HalA) (73),
plantaracin W (PlwA) (39), staphylococcin C55 (SacA) (80), lacticin 3147 (LtnA) (28), BHT (BhtA) (45),
Smb (SmbA) (131), and the two peptides of cytolysin (CylSA and CytLA) (24). The gene names used
here are as found in the databases, with the exception of SmbA1 (annotated as SmbB) and SmbA2
(annotated as SmbA). Ser/Thr residues that are not dehydrated are underlined. The LanT protease
cleavage sequences are shown in red; for cases in which additional proteolysis takes place (plantaricin W,
cytolysin, and haloduracin), the N terminus of the mature products is denoted with a red arrow.

486 Willey · van der Donk

 ANRV322-MI61-23 ARI 6 August 2007 18:7
residue of both peptides was mutated to Ala
provides a blueprint to better understand
these molecular interactions (19).
Enterococcal cytolysin is unusual in a
number of ways: Not only does it possess bac-
teriocin function, but it is cytolytic against
erythrocytes and other eukaryotic cells (24).
Although its structure has not been rigor-
ously established, sequence comparison with
lacticin 3147 and haloduracin (Figure 5)
suggests that both of its peptides contain
a MeLan at the N terminus and an addi-
tional internal Lan ring. The topology of
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org
the putative C-terminal ring of the CylLS
peptide remains to be resolved. The CylLL
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and CylLS prepropeptides are modified by
CylM and exported by a dual-function trans-
porter/protease to generate the CylLL  and
CylLS  peptides (24). A second round of pro-
teolysis by an extracellular serine protease,
CylA, is required for complete peptide matu-
ration, removing an identical six-residue se-
quence from the N terminus of each pep-
tide to yield CylLL  and CylLS  (24). The
β-peptides of the two-component lantibiotics
plantaricin W and haloduracin appear to also
require a second round of proteolytic process-
ing (39, 73); however, the responsible pro-
teases have not yet been identified.
Class III Lantibiotics: SapB and SapT
Both SapB and SapT are 21 residues
long, hydrophobic, and highly surface ac-
tive (Figure 3). These peptides are capa-
ble of self-assembly at air-water interfaces;
this is thought to provide a film of surfac-
tant through which nascent aerial filaments
emerge (105). Both peptides appear to be
well suited for residence at an air-water in-
terface, as modeling predicts that the macro-
cyclic structures would limit internal hydro-
gen bonding within polar side chains and force
the nonpolar side chains to the surface of the
molecule, thereby generating hydrophilic and
hydrophobic patches. These peptides are dis-
tinctly amphiphilic; the polar backbone atoms
may participate in extensive hydrogen bond-
ing with the surrounding water molecules,
and the hydrophobic side chains project out of
the water layer, contributing to the hydropho-
bicity that characterizes streptomycete aerial
structures and spores (55).
Among the four orthologues of the SapB
structural gene that have been identified, the
Cys and Ser residues that form both Lan
bridges and the Ser residues that remain as
Dha in mature SapB are conserved, suggesting
that these elements are important for activity.
The hydrophobicity of the two rings is also
conserved (128), so although the nature of the
interaction between peptides and the cell wall
remains obscure, it appears that amphiphilic-
ity and hydrophobicity are important.
Posttranslational Modification
Enzymes and Engineering Studies
The enzyme reaction mechanisms responsi-
ble for the dehydration and cyclization reac-
tions common to all lantibiotics are intriguing
because of their biochemistry and potential
applications to peptide engineering. The de-
hydration enzymes involved in class I (LanB)
and class II (LanM) lantibiotics demonstrate
low substrate specificity. For instance, NisB
dehydrates nonlantibiotic peptides fused to
the leader peptide of NisA expressed in L.
lactis (53, 91) and LctM can do the same in
vitro with peptides containing both proteino-
genic and nonproteinogenic amino acids (11).
LctM dehydration involves initial phospho-
rylation of the targeted Ser/Thr, followed by
elimination of the phosphate (10). The en-
zyme is processive with respect to the dehy-
dration reaction, completing all four dehy-
drations without releasing the substrate (75).
Although the leader peptide is required for the
correct action of LanB and LanM proteins, it
can be appended at its N terminus with pep-
tide tags (101, 130), including the export pep-
tides from the Sec and TAT secretion path-
ways (59), without affecting posttranslational
modification.
The LanC cyclization enzymes show
weak but clear sequence homology to the
www.annualreviews.org • Lantibiotic Function and Diversity 487
 ANRV322-MI61-23
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org
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ARI 6 August 2007 18:7
C-terminal domains of LanM proteins (97),
including one His and two Cys residues that
function as zinc ligands in the subtilin cyclase
SpaC (83). Recent elucidation of the crystal
structure of NisC shows the zinc on top of
an α,α-barrel that resembles the β-subunit of
protein farnesyl transferase (66). In addition
to the three protein ligands, the coordination
sphere is completed by a water molecule be-
lieved to be displaced by a Cys in the sub-
strate (66). The thiolate is then electrostati-
cally activated by the electron-rich metal site
toward attack onto the dehydro amino acid.
The active site consists of a shallow groove
that accommodates the formation of rings of
different sizes, but how NisC controls the re-
gioselectivity of cyclization remains unclear.
Biomimetic studies have demonstrated that
the enzyme must overcome the higher re-
activity of Dha compared with that of Dhb
in this process (134). Like the dehydration
carried out by NisB and LctM, the cycliza-
tion reaction catalyzed by NisC and LctM
has low substrate specificity, tolerating both
nonlantibiotic peptides (11, 53) and cysteine
derivatives (11).
Among other modification enzymes, the
oxidative decarboxylases that incorporate the
AviCys and MeAviCys structures at the C ter-
mini of epidermin and mersacidin (EpiD and
MrsD, respectively) have recently been crys-
tallized. They catalyze the oxidation of the C-
terminal Cys to the corresponding thioalde-
hyde followed by decarboxylation (3, 63).
EpiD also has low substrate specificity, as it
can oxidize a variety of synthetic peptides (95).
The mechanism by which l-Ser is converted
to d-Ala residues in lacticin 3147 was recently
reported (23). As predicted (98), this is a two-
step process involving dehydration of l-Ser
to Dha by peptide-specific LtnM enzymes
followed by stereospecific hydrogenation by
a single dehydrogenase, LtnJ, to yield d-Ala
(23).
New studies on lantibiotic engineering for
mersacidin (26, 104), lacticin 3147 (19, 20,
23), nukacin ISK-1 (1, 79), and lacticin 481
(11) have been reported; for results prior to
488 Willey · van der Donk
2003 we refer the reader to several compre-
hensive reviews (12, 21, 61). These investiga-
tions show the importance of Glu17 for mer-
sacidin (104) and the Lan and MeLan rings
in lacticin 481 for antimicrobial activity. In-
terconversion of Lan and MeLan is tolerated
by the biosynthetic machinery with relatively
little influence on the antimicrobial activities
of lacticin 481 (11) and mutacin II (14). How-
ever, such substitution results in a large de-
crease in activity for nisin (61). Removal of
the N-terminal five residues from lacticin 481
(110) and the three N-terminal lysines from
the related compound nukacin ISK-1 (1) re-
sults in a 10-fold and 32-fold reduction of
bioactivity, respectively, indicating the impor-
tance of the positively charged N terminus. In
addition to these investigations, several recent
engineering studies have focused on increas-
ing production of wild-type lantibiotics (13,
20, 35).
LANTIBIOTIC GENETICS
The recent increase in the number of cloned
lantibiotic biosynthetic genes has revealed
several trends, including the presence of reg-
ulatory and immunity genes in addition to
biosynthetic operons and the regulated ex-
pression of these genes in a growth-phase-
dependent fashion. As more gene clusters are
sequenced, the potential for identifying new
lantibiotics through genomic approaches is
becoming realized (29, 64, 73).
Biosynthetic Gene Clusters
The genes for lantibiotic biosynthesis are gen-
erally found in clusters, which can be chro-
mosomal, on transposable elements, or on
plasmids, raising questions about the impor-
tance of horizontal gene transfer. In many
cases the lanA structural gene is the first gene
in an operon that includes genes encoding
the modification enzymes (lanB and lanC or
lanM ) and exporter (lanT ) (Figure 6). For
those lantibiotics that undergo other, more
unusual posttranslational modifications, the
genes encoding the required enzymes are
 ANRV322-MI61-23 ARI 6 August 2007 18:7

Nisin
L. lactis
A B T C I P R K F E G

Subtilin
B. subtilis B T C S I F E G R K

Epidermin
S. epidermis A
G E F H T B C D Q P

Pep 5 I A
S. epidermis T P B C

Lacticin 481
L. lactis A
M T F E G
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org

Cinnamycin
by Pennsylvania State University on 05/22/12. For personal use only.

S. cinnamoneus
3 4 7 A M X T H Y Z 8 9 R K 10 11 12 14

13
Mersacidin R1
B. subtilis K2 R2 F G E A R1 D M T

Lacticin 3147 E F I R A1 A2 M1 T M2 J
L. lactis

SapB ramC ramS ramA ramB ramR

S. coelicolor
SapB

Figure 6
Representative biosynthetic gene clusters of selected lantibiotics. Transcriptional units, where known, are
indicated by red arrows. Dashed arrow represents a putative promoter, the exact location of which has not
yet been determined (37). Note that the gene nomenclature differs for SapB for historical reasons (128).

usually found in this cluster. Recent exam- Regulation of Lantibiotic Synthesis

ples include ltnJ and cinX, which encodes the
In almost all cases, lantibiotic production
hydroxylase that oxidizes the Asp residue of
is coordinately regulated with other cellu-
cinnamycin both in vivo (M.J. Bibb, personal
lar events. In addition, biosynthesis of a
communication) and in vitro (E. Fogle & W.A.
number of lantibiotics is autoregulatory with
van der Donk, unpublished results). How-
transcription of biosynthetic and immunity
ever, this is not always the case; mrsD is con-
operons governed by two-component regu-
vergently transcribed relative to other mer-
latory systems. Many of these autoregulatory
sacidin biosynthetic enzymes. Other genes
mechanisms have been described as quorum-
involved in regulation and/or immunity may
sensing systems (32, 60, 100).
be within the biosynthetic operon or, more
Autoregulation of nisin and subtilin in-
commonly, found in closely linked operons.
volves subinhibitory concentrations of extra-
For instance, the biosynthetic gene cluster re-
cellular lantibiotics, which trigger the activity
sponsible for cinnamycin production includes
of a signal kinase/response regulator signal
17 genes under the control of nine promoters
transduction system. This results in the tran-
(123; S. O’Rourke, personal communication)
scription of biosynthetic and immunity genes,
(Figure 6).

www.annualreviews.org • Lantibiotic Function and Diversity 489

 ANRV322-MI61-23 ARI 6 August 2007 18:7

with peptide production beginning during
mid-exponential growth and peaking at the
log-phase to stationary-phase transition (60,
100). The nisin and subtilin autoregula-
tory systems have inspired the construction
of lantibiotic-specific tools for heterologous
gene expression: the so-called NICE (nisin-
controlled gene expression) (61) and SURE
(subtilin-regulated gene expression) systems
(5).
Advances in B. subtilis genetics have re-
vealed a more complex regulatory network
regulating subtilin biosynthesis. The cell in-
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org
otic production, and sporulation. Subtilin
biosynthesis depends on transcription of the
spaRK operon, which encodes the response
regulator and signal kinase. This operon is
in turn dependent on the alternative sigma
factor, σH , which is regulated at both the
transcriptional and translational levels. For
instance, sigH transcription is inhibited by
the transition state regulator AbrB (100)
(Figure 7).
Although Bacillus sp. HILY-85 also seems
to coordinately regulate mersacidin biosyn-
thesis with other stationary-phase events,

tegrates multiple environmental signals prior it is not produced until stationary phase
to initiating a suite of late-log- and stationary- commences, and it is σH independent (31),
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phase behaviors such as competence, antibi- as recently demonstrated in an engineered

Figure 7
Presubtilin is the product of the spaS gene; it is modified and transported by the membrane-associated
subtilin synthetase complex (52), which consists of SpaB, SpaC, and SpaT. After export, the leader is
cleaved by any one of three exoproteases (subtilisin, WprA, Vpr) (18). Mature subtilin serves as ligand for
the sensor kinase SpaK, which when activated phosphorylates SpaR, a positive regulator of spaS and the
biosynthetic and immunity operons. Stationary-phase control of spaS transcription is regulated indirectly
through σH control of spaRK; other genes in the cluster are dependent on σA for transcription. σH itself
is subject to transcriptional control by the transition state regulator AbrB (100).

490 Willey · van der Donk

 ANRV322-MI61-23 ARI 6 August 2007 18:7

mersacidin expression system (26). Two dif-

ferent response regulators separately govern
self-immunity and mersacidin biosynthesis:
MrsR2 and its cognate sensor kinase, MrsK2,
regulate immunity, and MrsR1 governs
biosynthesis. The kinase that phosphory-
lates MrsR1 has not yet been identified, and
as mersacidin does not appear to be au-
toregulatory, the signal that triggers its pro-
duction remains unknown (31). Mersacidin
is not the only lantibiotic whose synthe-
sis is controlled, at least in part, by a so-
called orphan response regulator; lacticin
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org

3147 (72), mutacin II (87), epidermin (85),

and SapB (49, 81, 82) are also subject to such
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regulation.
S. mutans is a principal component of oral
biofilms, where interspecies competition is
presumed to be intense owing to limited re-
sources and high biodiversity. This would
make antibiotic production an adaptive ad-
vantage, and indeed mutacin I production is
stimulated when S. mutans is grown on solid
substrates (88). Its production is regulated in
concert with other cellular activities includ-
ing competence, biofilm formation, and acid
tolerance (74, 121). A complex regulatory net-
work is emerging that involves the autoinduc-
ing signal LuxS, which is needed for efficient
transcription of the mutA structural gene and,
to a lesser extent, of mutR; a transcriptional
repressor, irvA, which itself is negatively reg-
ulated by LuxS (74, 107); and a sensor kinase
(CiaH) (89).
Simultaneous bacteriocin production and
competence development might provide a
mechanism to ensure a pool of homologous
DNA for uptake by selectively targeting non-
competent cells of the same strain (56). Smb
production by S. mutans GS5 is coregulated
with competence, as Smb production depends Figure 8
on the quorum-sensing signal competence- (a) In the absence of a target microbe or erythrocyte, CylR2 binds to the
stimulating peptide (CSP) encoded by the promoter that governs cytolysin biosynthetic genes. However, basal
transcription enables a small amount of mature CylL  and CylS  to
comC gene. A com box-like element is present accumulate. Under these circumstances, CylL  titrates CylS  . (b) In the
upstream of the smbA structural gene (131), presence of a target cell, CylL  preferentially binds the lipid bilayer. CylS 
and an S. mutans GS5 comC mutant that lacks is thus free to (i) interact with the membrane-bound CylR1, which leads to
antibiotic production activity is rescued by derepression of the cytolysin biosynthetic operon, and (ii) bind to CylL
the addition of synthetic CSP to the medium localized to the target cell. Both peptides are needed for cytolytic activity.

www.annualreviews.org • Lantibiotic Function and Diversity 491

 ANRV322-MI61-23
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by Pennsylvania State University on 05/22/12. For personal use only.
ARI 6 August 2007 18:7
(132). An 18-residue CSP was recently puri-
fied from S. mutans GS5; apparently cleavage
of the three C-terminal amino acids by an un-
known extracellular protease potentiates CSP
activity (86).
The biosynthesis of epidermin by S. epi-
dermidis is, at least in part, also controlled by
global regulators of cellular stress response
and biofilm production (51). Transcriptional
regulation of epiA is governed by the response
regulator EpiQ (85); however, peptide matu-
ration is under the control of the global reg-
ulatory system, agr (accessory gene regula-
tor) (46, 47). When the agr quorum system is
active, many surface proteins are downregu-
lated while many exoproteins are upregulated
(118). Epidermin activity is thus controlled
via the agr-regulated protease EpiP, which
cleaves the epidermin leader peptide (51).
The cell-density-dependent mechanism
that regulates the production of the two-
component lantibiotic cytolysin reflects its
role as an Enterococcus faecalis virulence fac-
tor (Figure 8). In the absence of target cells,
cytolysin production is repressed by CylR1,
which dimerizes and binds specifically to an
inverted repeat that overlaps the –35 re-
gion of the cytolysin operon promoter (93).
However, basal transcription of the biosyn-
thetic operon results in low-level produc-
tion of CylLS  and CylLL  , which form an
insoluble complex that has neither cytolytic
nor regulatory activity. When target cells are
present, CylLL  preferentially binds to phos-
phatidylcholine:cholesterol lipid bilayers and
can no longer titrate free CylLS  , whose ac-
cumulation results in high-level cytolysin ex-
pression (17). Although it is not known how
CylLS  causes derepression, a second, mem-
SUMMARY POINTS
1. Lantibiotics are translated as prepropeptides that are first modified by the dehydration
of Ser and Thr residues in the C-terminal half of the peptides. Lan or MeLan bridges
are then introduced by the intramolecular addition of Cys thiols to the dehydrated
amino acids. The peptides do not gain full biological function until the leader sequence
is cleaved.
492 Willey · van der Donk
brane binding protein of unknown function,
CylR2, is also involved (24). This economy of
regulation allows Enterococcus faecalis to use a
single peptide to probe the environment for
cytolysin targets and induce its production
only when appropriate.
Extracellular pH regulates the production
of lacticin 481 by strains of L. lactis. The lac-
ticin 481 biosynthetic operon (lctAMTFEG)
is governed by two promoters, P1 and P3,
which map upstream of lctA. Acidification of
the medium by lactic acid stimulates expres-
sion from both promoters, and while the P3
promoter appears to be the stronger of the
two, P1 is more tightly regulated (37). The
P1 promoter features a novel 14-bp regulatory
region to which the positive regulatory pro-
tein RcfB binds. RcfB is a member of the Crp-
Fnr family of transcriptional activators and is
cotranscribed with a universal stress-like pro-
tein and a multidrug transporter. RcfB null
mutants are significantly diminished in their
ability to survive acidic conditions, suggesting
that RcfB is involved in regulating multiple
genes whose expression is required for acid
tolerance (67).
CONCLUSIONS AND OUTLOOK
The past decade has seen great leaps in our un-
derstanding of the mode of action and biosyn-
thesis of lantibiotics, and it has revealed that
this class of molecules is not limited to antimi-
crobial peptides. The list of new lantibiotics
and potential applications continues to expand
and the immediate future is likely to see new
breakthroughs in the discovery, engineering,
and mode of action of lantibiotics (see Future
Issues, below).
 ANRV322-MI61-23 ARI 6 August 2007 18:7

2. A new classification scheme is proposed: Class I peptides are modified by separate

dehydratases (LanB) and cyclases (LanC). Class II lantibiotics are dehydrated and
cyclized by a single LanM-type enzyme. Class III peptides have little or no antibiotic
activity; rather, they provide alternative physiological functions.
3. Most lantibiotics exert their antibiotic effect by either forming pores in the target
cell membrane or inhibiting cell wall synthesis. Importantly, there is little evidence of
stable resistance mutants arising in food products that have been treated with nisin,
which has been used for almost 50 years.
4. In vitro reconstitution of LanM-type enzymes reveals that the Ser and Thr residues
are first phosphorylated prior to elimination, and both in vitro and in vivo studies
show that the biosynthetic enzymes have remarkable substrate promiscuity. The latter
Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org

findings are promising for lantibiotic engineering.

5. The genes for lantibiotic biosynthesis, regulation, and self-immunity are found in
by Pennsylvania State University on 05/22/12. For personal use only.

clusters. Lantibiotic biosynthesis is frequently coregulated as part of a stress response

when cells enter late-log or stationary phase.

FUTURE ISSUES
1. As new lantibiotic peptides are discovered, it is most likely that new structural elements
and functionalities will be revealed.
2. New clinical applications of a variety of lantibiotics are currently being pursued,
underscoring their potential as important agents in disease control.
3. Protein engineering is readily performed in these ribosomally synthesized peptides,
and thus lantibiotics are especially amenable to rational drug design.
4. Database mining and PCR analysis of genomic DNA could greatly expand the number
and diversity of currently known lantibiotics.
5. The mode of action of lantibiotics such as Pep5, lacticin 481, and epilancin that are
structurally unrelated to nisin, mersacidin, or cinnamycin will be the subject of further
investigation.

DISCLOSURE STATEMENT
The authors are not aware of any biases that might be perceived as affecting the objectivity of
this review.

ACKNOWLEDGMENTS
The authors would like to thank Mr. Alex Smiros for his assistance in preparing the figures.
The authors also acknowledge support by the National Institutes of Health; WAD is supported
by NIH R01 GM58822, and JMW by NIH R15 GM069398.

www.annualreviews.org • Lantibiotic Function and Diversity 493

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Annual Review of
Microbiology

Volume 61, 2007 Contents

Frontispiece
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Margarita Salas p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p xiv

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40 Years with Bacteriophage Ø29

Margarita Salas p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 1
The Last Word: Books as a Statistical Metaphor for Microbial Communities
Patrick D. Schloss and Jo Handelsman p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 23
The Mechanism of Isoniazid Killing: Clarity Through the Scope
of Genetics
Catherine Vilchèze and William R. Jacobs, Jr. p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 35
Development of a Combined Biological and Chemical Process for
Production of Industrial Aromatics from Renewable Resources
F. Sima Sariaslani p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 51
The RNA Degradosome of Escherichia coli: An mRNA-Degrading
Machine Assembled on RNase E
Agamemnon J. Carpousis p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 71
Protein Secretion in Gram-Negative Bacteria via the Autotransporter
Pathway
Nathalie Dautin and Harris D. Bernstein p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 89
Chlorophyll Biosynthesis in Bacteria: The Origins of Structural and
Functional Diversity
Aline Gomez Maqueo Chew and Donald A. Bryant p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p113
Roles of Cyclic Diguanylate in the Regulation of Bacterial Pathogenesis
Rita Tamayo, Jason T. Pratt, and Andrew Camilli p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p131
Aggresomes and Pericentriolar Sites of Virus Assembly:
Cellular Defense or Viral Design?
Thomas Wileman p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p149
As the Worm Turns: The Earthworm Gut as a Transient Habitat for
Soil Microbial Biomes
Harold L. Drake and Marcus A. Horn p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p169

vi
 AR322-FM ARI 9 July 2007 9:23

Biogenesis of the Gram-Negative Bacterial Outer Membrane

Martine P. Bos, Viviane Robert, and Jan Tommassen p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p191
SigB-Dependent General Stress Response in Bacillus subtilis and
Related Gram-Positive Bacteria
Michael Hecker, Jan Pané-Farré, and Uwe Völker p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p215
Ecology and Biotechnology of the Genus Shewanella
Heidi H. Hau and Jeffrey A. Gralnick p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p237
Nonhomologous End-Joining in Bacteria: A Microbial Perspective
Robert S. Pitcher, Nigel C. Brissett, and Aidan J. Doherty p p p p p p p p p p p p p p p p p p p p p p p p p p p p259
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Postgenomic Adventures with Rhodobacter sphaeroides

Chris Mackenzie, Jesus M. Eraso, Madhusudan Choudhary, Jung Hyeob Roh,
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Xiaohua Zeng, Patrice Bruscella, Ágnes Puskás, and Samuel Kaplan p p p p p p p p p p p p p p p p p283
Toward a Hyperstructure Taxonomy
Vic Norris, Tanneke den Blaauwen, Roy H. Doi, Rasika M. Harshey,
Laurent Janniere, Alfonso Jiménez-Sánchez, Ding Jun Jin,
Petra Anne Levin, Eugenia Mileykovskaya, Abraham Minsky,
Gradimir Misevic, Camille Ripoll, Milton Saier, Jr., Kirsten Skarstad,
and Michel Thellier p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p309
Endolithic Microbial Ecosystems
Jeffrey J. Walker and Norman R. Pace p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p331
Nitrogen Regulation in Bacteria and Archaea
John A. Leigh and Jeremy A. Dodsworth p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p349
Microbial Metabolism of Reduced Phosphorus Compounds
Andrea K. White and William W. Metcalf p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p379
Biofilm Formation by Plant-Associated Bacteria
Thomas Danhorn and Clay Fuqua p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p401
Heterotrimeric G Protein Signaling in Filamentous Fungi
Liande Li, Sara J. Wright, Svetlana Krystofova, Gyungsoon Park,
and Katherine A. Borkovich p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p423
Comparative Genomics of Protists: New Insights into the Evolution
of Eukaryotic Signal Transduction and Gene Regulation
Vivek Anantharaman, Lakshminarayan M. Iyer, and L. Aravind p p p p p p p p p p p p p p p p p p p p453
Lantibiotics: Peptides of Diverse Structure and Function
Joanne M. Willey and Wilfred A. van der Donk p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p477
The Impact of Genome Analyses on Our Understanding of
Ammonia-Oxidizing Bacteria
Daniel J. Arp, Patrick S.G. Chain, and Martin G. Klotz p p p p p p p p p p p p p p p p p p p p p p p p p p p p p503

Contents vii
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Morphogenesis in Candida albicans

Malcolm Whiteway and Catherine Bachewich p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p529
Structure, Assembly, and Function of the Spore Surface Layers
Adriano O. Henriques and Charles P. Moran, Jr. p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p555
Cytoskeletal Elements in Bacteria
Peter L. Graumann p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p589

Indexes

Cumulative Index of Contributing Authors, Volumes 57–61 p p p p p p p p p p p p p p p p p p p p p p p p619

Annu. Rev. Microbiol. 2007.61:477-501. Downloaded from www.annualreviews.org

Cumulative Index of Chapter Titles, Volumes 57–61 p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p622

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Errata

An online log of corrections to Annual Review of Microbiology articles may be found

at http://micro.annualreviews.org/

viii Contents