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Lysine decarboxylase

Related terms:

 Agar

 Lactose

 Glutamic acid

 Lysine

 Arginine

 Ornithine decarboxylase

 Gamma-Aminobutyric acid

 Antiporter

 Serotype

 Glucose

Learn more about Lysine decarboxylase

SALMONELLA | Detection

T. Humphrey, P. Stephens, in Encyclopedia of Food Sciences and Nutrition (Second Edition), 2003

Confirmatory Tests

Other organisms, particularly members of the family Enterobacteriaceae, can resemble Salmonella on
selective agars and confirmation of ‘Salmonella-like’ colonies are necessary. Tests take the form of
biochemical confirmation, which usually involves assessment of urease and lysine
decarboxylase activity, fermentation of dulcitol, indole production, growth in the presence of potassium
cyanide, utilization of sodium malonate and, more recently, pyrrolindonylarylamidase (PYR) activity.
These reactions, in combination with serological tests, are usually sufficient for identification, but
additional tests may sometimes be necessary. If these are to form part of a laboratory routine, the use
of commercial kits may be cost-effective. Antibodies against somatic (O) and flagella (H) antigens are
used to confirm/identify Salmonella-like isolates. Somatic antigens are composed of polysaccharide,
while those from the flagella are proteinaceous. Testing will usually comprise slide agglutinations with
polyvalent O and H antisera, followed by the use of sera raised against specific antigens.

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Etiologic Agents of Infectious Diseases

Theresa J. Ochoa, Miguel O’Ryan, in Principles and Practice of Pediatric Infectious Diseases (Fourth
Edition), 2012
Yersinia are gram-negative, aerobic and facultatively anaerobic, rod-shaped nonspore-forming bacteria;
these microorganisms ferment glucose, are oxidase-negative, and reduce nitrates to nitrites. Yersinia do
not react with hydrogen sulfide, citrate, potassium cyanide (KCN), lysine decarboxylase (LDC), and
arginine dihydrolase, but do react with methyl red and indole. Yersinia species are nonpigmented, are
nonlactose-fermenting, and produce acid from glucose. Species are differentiated by a variety of traits,
such as urease (Y. pestis is negative), motility (Y. pestis is nonmotile at both 25°C and 37°C), ornithine
decarboxylase (Y. enterocolitica is positive), and rhamnose (Y. pseudotuberculosis is positive), as well as
several carbohydrate fermentation reactions for which Y. enterocolitica is positive.

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Salmonella

Steven L. Percival∗, David W. Williams∗∗, in Microbiology of Waterborne Diseases (Second Edition), 2014

Metabolism and Physiology

Salmonellae are facultative anaerobes and are catalase positive, oxidase negative and ferment glucose,
mannitol and sorbotol to produce acid or acid and gas. Whilst S. arizonae is able to ferment lactose, this
is the exception rather than the rule. As a group, Salmonella are able to ferment sucrose, but rarely
adonitol and overall do not form indole. They also do not hydrolyze urea or deaminate phenylalanine,
but usually form H2S on triple sugar iron agar and can use citrate as a sole carbon
source. Salmonella form lysine and ornithine decarboxylases, exceptions to this include S. paratyphi A
and S. typhi. Salmonellae yield negative Voges-Proskauer and positive methyl red tests and do not
produce cytochrome oxide. Salmonellae are also unable to deaminate tryptophan or phenylalanine and
are usually urease and indole negative. Based on the biochemical tests above, Salmonella can
presumptively be identified. However, for a more detailed identification of Salmonella, isolates are
generally serotyped, especially for epidemiological investigations. As mentioned previously, typing
of Salmonella is based on the recognition of bacterial surface antigens: the thermostable polysaccharide
cell wall or somatic (‘O') antigens and the thermolabile flagella proteins or ‘H' antigens. It is also possible
to subtype Salmonella serotypes on the basis of phage typing. Sub-divisions of Salmonella can also be
undertaken by plasmid profiling, ribotyping or pulsed field gel electrophoresis (PFGE) of DNA fragments
generated from restriction enzyme digestion.

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Physiology of Prokaryotic Cells

Dennis W. Grogan, in Cell Physiology Source Book (Fourth Edition), 2012

VIB

Acid Stress

Other responses to the environment involve changes in the pattern of gene expression which are often
complex and lead to corresponding changes in the protein composition of the prokaryotic cell
(Neidhardt et al., 1990). The response of enteric bacteria to pH stress provides an example of this
strategy that involves changing the amount of enzymes and other proteins. When the pH of the growth
medium decreases sharply, E. coliand Salmonella enterica increase the synthesis rate of about 50
proteins. These acid-induced proteins include catabolic (degradative) enzymes that decarboxylate basic
amino acids (arginine, ornithine and lysine decarboxylases). The action of these enzymes on amino acids
taken up from the environment generates CO2 and diamines; the latter are exported from the cell and
act to neutralize the acidic growth medium. Similarly, acid stress during anaerobic growth on a
fermentable sugar increases lactate dehydrogenase and formate-hydrogen lyase; both of these enzymes
divert the flow of intermediates through fermentative metabolism in ways that decrease the amount of
acid produced (Slonczewski and Foster, 1996).

Many other prokaryotic responses to environmental change similarly involve changing the protein
composition of the cell by modulating transcription of certain genes. This general strategy takes
advantage of the relatively large biosynthetic capacity of the growing prokaryotic cell, its simple
organization and a very short half-life of messenger RNA which, in E. coli (under standard conditions)
averages 1.3 min. Studies of specific responses in bacteria have revealed an array of elegant and diverse
molecular mechanisms for controlling gene expression, which lie outside the scope of this chapter. One
example will be described, however, which involves transmembrane signaling and appears in many
environmental responses in bacteria.

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Shigella and enteroinvasive Escherichia coli

Anthony T. Maurelli, in Escherichia coli (Second Edition), 2013

Classification and biochemical characteristics

The Japanese microbiologist Shiga isolated an organism (Shigella dysenteriae) from the dysenteric stool
of a stricken individual in 1898. Three more Shigella species (S. boydii, S. flexneri, and S. sonnei) were
subsequently identified and grouped by serotype and metabolic activities. The four species of the
genus Shigellaare grouped serologically (41 serotypes) based on their somatic O-antigens: S.
dysenteriae (group A), S. flexneri (group B), S. boydii(group C), and S. sonnei (group D). As members of
the family Enterobacteriaceae, they are closely related to the salmonellae (Ochman et al.,
1983). Shigella are non-motile, Gram-negative rods. Some important biochemical characteristics that
distinguish these bacteria from other enterics are their inability to utilize citric acid as a sole carbon
source and their inability to ferment lactose, although some strains of S. sonnei may ferment lactose
slowly. They are oxidase-negative, do not produce H2S (except for S. flexneri 6 and S. boydii serotypes 13
and 14), and do not produce gas from glucose. Shigella spp. are inhibited by potassium cyanide and do
not synthesize lysine decarboxylase(Ewing, 1986).

Based on the cumulative sequence evidence from 16S rRNA genes, multiple housekeeping genes, and
whole genome comparisons, the pathogenic Shigella can be considered a subset of non-pathogenic E.
coli (Cilia et al., 1996; Pupo et al., 1997; Shu et al., 2000; Jin et al., 2002). Although phylogenetically it
would be more accurate to treat Shigella as pathotypes of E. coli, the members of the
genus Shigella continue to be divided for historical and medical purposes into the four species or
subgroups described above (Strockbine and Maurelli, 2005).

Enteroinvasive E. coli (EIEC) isolates that cause a diarrheal illness identical to Shigella dysentery were
identified in the 1970s (DuPont et al., 1971; Tulloch et al., 1973). The pathogenic and biochemical
properties that EIEC share with Shigella pose a problem for distinguishing these pathogens. For example,
unlikenormal flora E. coli, EIEC are non-motile and 70% of isolates are unable to ferment lactose (Silva et
al., 1980; van den Beld and Reubsaet, 2012). These are also features of Shigella. More striking is the
observation that strains of EIEC are almost universally negative for lysine decarboxylase (LDC) activity
whereas almost 90% of normal flora E. coli are positive. In this respect, EIEC also
resemble Shigella, which uniformly lack LDC activity. Some serotypes of EIEC even share identical O-
antigens with Shigella (Sansonetti et al., 1985). By contrast, EIEC resemble E. coli in their ability to
ferment xylose and to produce gas from glucose, both traits for which Shigella are negative (Silva et al.,
1980).

It is now generally accepted that the present day strains of Shigella arose multiple times from as many
as seven independent ancestral strains of E. coli (Pupo et al., 2000; Yang et al., 2005). EIEC probably
evolved later than Shigella and from different E. coli ancestors (Lan et al., 2004). However, the common
seminal event in the evolution of both groups of pathogens was the acquisition of a large plasmid that
encodes the genes necessary for invasion of mammalian cells (see below) (Lan et al., 2001).

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Etiologic Agents of Infectious Diseases

Dwight A. Powell, Mario J. Marcon, in Principles and Practice of Pediatric Infectious Diseases (Fourth
Edition), 2012

Microbiology and Epidemiology

The genus Citrobacter has undergone significant taxonomic revision through the use of newer
techniques based on DNA relatedness. The genus now contains 11 named species: Citrobacter freundii,
C. koseri, C. amalonaticus, C. youngae, C. farmeri, C. braakii, C. werkmanii, C. sedlakii, C. gillenii, C.
murliniae,and C. rodentium.1–5 Citrobacter koseri has replaced the taxon formerly known as Citrobacter
diversus, and Citrobacter farmeri is the new taxon assigned to the former Citrobacter
amalonaticusbiogroup 1. All species except C. rodentium have been recovered from human clinical
sources (some rarely) including blood and other normally sterile body sites, wounds, respiratory and
urinary tract; however, C. freundii and C. koseri are the most important human pathogens. C. freundii, C.
koseri, and C. amalonaticusappear to be distinct organisms; however, only C. koseri appears to be
genetically homogeneous. Several other named species form a closely related group and are difficult to
differentiate biochemically; they are sometimes referred to as C. freundii-complex organisms.

Members of the genus Citrobacter share all the general properties and biochemical characteristics of the
family Enterobacteriaceae, including the following: gram-negative rod, catalase-positive and oxidase-
negative, growth on MacConkey agar, reduction of nitrate to nitrite, growth both aerobically and
anaerobically, and fermentation of glucose and other carbohydrates. Most isolates are motile and utilize
citrate as a sole carbon source, but lack urease and lysine decarboxylaseactivity; production of hydrogen
sulfide is variable, occurring with C. freundii and a few other species. On salmonella-shigella agar,
lactose-negative/hydrogen-sulfide positive isolates of Citrobacter spp. produce black colonies
resembling Salmonellaspp. The lysine decarboxylase reaction allows for separation of the hydrogen
sulfide-producing isolates of Citrobacter spp. from Salmonella spp.1–5 Select isolates of C. freundii have
“O” (somatic) cell wall antigens closely related to the O antigens of Salmonellaspp. and thus cross-react
with Salmonella typing antisera.5 In addition, rare isolates of C. freundii and C. braakii cross-react with
some commercial Escherichia coli O157 typing antisera.5 For this reason, it is always prudent to confirm
the identification of suspected Salmonella spp. and E. coli O157 by the use of both biochemical and
serologic methods. Although biochemical identification of the newer species can be accomplished
through the use of conventional tests, most commercial identification systems do not include all species
in their databases. This limitation hampers the correlation of the newer species with human disease.

Citrobacter spp. are primarily inhabitants of the intestinal tract of mammals and other vertebrates. Their
isolation from environmental sources such as water and soil likely is the result of fecal
excretion. Citrobacter spp. are not common agents of human disease, and are most often recovered
from stool as colonizing flora of the gastrointestinal tract. When associated with significant human
infection, Citrobacter can be recovered from blood, cerebrospinal fluid (CSF), urine, respiratory tract
secretions, and wounds. The most common Citrobacter spp. isolated from human sources are C.
freundii (all sites listed above), C. koseri (all sites but CSF and brain most commonly), C. amalonaticus (all
sites except CSF), C. braakii (primarily stool), and C. youngae (primarily stool).

The pathogenesis of Citrobacter infections has not been characterized fully. Most C. koseri isolates
produce hemolysins, are piliated, and are resistant to killing by pooled human sera. Tropism for the
central nervous system may be due to specific outer membrane proteins. In one study, 79% of strains
of C. koseri isolated from CSF had a unique 32-kd outer-membrane protein, which was found in only 9%
of isolates from other kinds of specimens.6 Citrobacter koseri also has the ability to enter macrophages,
survive phago-lysosomal fusion, and replicate intracellularly. These infected macrophages then infiltrate
blood vessels in the brain, which may be one of the main mechanisms of infecting brain microvascular
endothelial cells, thus starting the process leading to brain abscess.5,7,8

In the pediatric population, infections due to Citrobacter spp. occur most commonly in
neonates.9,10 Organisms can be transmitted by vertical transmission from mothers or by nosocomial
spread. Direct mother-to-infant transmission has been confirmed by ribotyping and DNA
fingerprinting.11,12 It is likely that individual strains of Citrobacter spp. circulating in the community
periodically gain access to a hospital nursery from the hands of nursery personnel and visitors.13 One
nursery outbreak of C. freundii was traced to contaminated infant formula.14 Outbreaks in nursery
settings can last for months or years and appear to be best controlled by cohorting of patients and
personnel.

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Etiologic Agents of Infectious Diseases

Katalin I. Koranyi, Mario J. Marcon, in Principles and Practice of Pediatric Infectious Diseases (Fourth
Edition), 2012

Edwardsiella Species

Organisms of the genus Edwardsiella are found in freshwater environments and cold-blooded animals,
including fish, reptiles, and amphibians, but occasionally also in birds and mammals. E. tarda, E.
hoshinae, and E. ictaluri are the three recognized species of this genus, but only E. tarda has been
associated with human disease.1–6 Edwardsiella generally are positive for both lysine and ornithine
decarboxylase activity, but yield a negative Voges–Proskauer test result and do not use citrate as a sole
carbon source. Most clinical isolates of E. tarda produce indole and hydrogen sulfide, but do not
ferment lactose so appear as colorless colonies on MacConkey agar. This characteristic, along with the
production of hydrogen sulfide on salmonella-shigella agar, necessitates further testing of such isolates
to exclude Salmonella spp.1–6 E. tarda is not considered to be part of the normal human intestinal flora,
being recovered in <1% of stool samples in studies of carriage. Thus human infections generally result
from exposure to the organism from its natural environments.

A variety of human infections due to E. tarda are described, but none has been associated with
outbreaks or epidemics of disease.11–19 Infections due to E. tarda can be divided into intestinal and
extraintestinal. The pathogenesis of gastroenteritis due to E. tarda is not well established. The organism
is negative for Shiga-like enterotoxin and in the Sereny test of entero-invasiveness, but it does produce
cell-associated hemolysins and siderophores and can penetrate HeLa and HEp-2 cells in
culture.17 Intestinal infections usually manifest as self-limiting gastroenteritis associated with
consumption of raw seafood or snake flesh or other exposure from aquatic environments. Evidence
supporting the association of E. tarda with gastroenteritis is largely circumstantial. E. tarda is isolated
from stool cultures significantly more frequently in individuals with diarrhea than in asymptomatic
individuals.1,2 The gastroenteritis typically is an acute secretory diarrheal illness, although serious cases
of dysentery can also occur. Intestinal infections in immunocompromised patients including transplant
patients can be severe.19 Reported extraintestinal infections include: wound infections such as cellulitis
and gas gangrene associated with trauma; septicemia and meningitis; and focal infections, including
cholecystitis, prosthetic valve endocarditis, hepatic abscesses, tubo-ovarian abscess, pyogenic arthritis,
and osteomyelitis.1,11–15

Wound infections are perhaps the most important infections caused by E. tarda. They usually occur after
local trauma, such as abrasion, laceration, or penetrating injury. Many infections have been associated
with aquatic injuries and represent combined infection due to E. tarda and Aeromonas
hydrophila.1 Pyogenic arthritis caused by E. tarda after catfish puncture wound has been reported.15 E.
tarda BSI is most commonly associated with an underlying condition. In adults, liver disease is a
common predisposing factor. In children, sickle-cell anemia has been linked to serious infections due
to E. tarda, as it has been with Salmonella spp.11 Osteomyelitis and meningitis have occurred in
individuals with hemoglobinopathies. The gastrointestinal tract is presumably the site of E.
tarda colonization in these patients, with invasion leading to BSI and subsequent focal infection.
Neonatal septicemia and meningitis due to E. tarda has been reported in association with acquisition
from mother.13

Typically, E. tarda is susceptible to ampicillin, cefazolin, third-generation cephalosporins, extended-

spectrum penicillin/inhibitor combination antibiotics, aminoglycosides, fluoroquinolones, and
trimethoprim-sulfamethoxazole. In serious invasive infections, combination therapy with an extended-
spectrum cephalosporin and an aminoglycoside has been successful. Antibiotic therapy of individuals
with gastroenteritis due to E. tarda probably is not indicated.

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Shigella and Shigellosis

Sophie Octavia, Ruiting Lan, in Molecular Medical Microbiology (Second Edition), 2015

Microbiology
Shigella can be distinguished from E. coli by their classic biochemical properties, including negative
lysine decarboxylation, non-lactose fermentation and non-motility [5]. To isolate Shigella from a clinical
sample, MacConkey and xylose-lysine-desoxycholate (XLD) agars are commonly used, with lactose in the
former and xylose and lysine in the latter media to differentiate it from E. coli (Table 65.1). However,
EIEC is often non-motile as well as lactose and lysine decarboxylase negative. Xylose fermentation is the
only property that may differentiate EIEC from Shigella in these media. To further differentiate EIEC
from Shigella, additional biochemical tests, such as utilization of l-serine and sodium acetate, and
mucate fermentation may be used [5]. Alternatively, salicin fermentation, aesculin hydrolysis, and the
combined positivity of gas from d-glucose and indole production can also be used [8]. EIEC is positive for
one or more of the these properties whereas Shigella is generally negative [9]. EIEC is generally more
biochemically active than Shigella.

Differentiation of Shigella from E. coli by molecular methods is not yet feasible despite attempts using
some gene targets [10]. However, Shigella and EIEC can be differentiated from other E. coli. The
gene ipaH (invasion plasmid antigen H), which is present on the virulence plasmid as well as on the
chromosome in multiple copies, is often used as the gene target [8,11] and offers good sensitivity [12].

The differentiation of the four species, S. dysenteriae, S. flexneri, S. boydii and S. sonnei, requires a
combination of biochemical tests and serological typing [5], since relatively few biochemical properties
are useful and absolute for separation of the species. Utilization of mannitol and decarboxylation of
ornithine are primarily used [13]. S. sonnei is positive for both while S. dysenteriae is negative for
both. S. flexneri and S. boydii are positive of mannitol but negative for ornithine, with the exception of S.
boydii serotype 13 which is ornithine-positive [5]. No biochemical traits can differentiate S.
flexneri from S. boydii. Serogrouping is essential for differentiation of S. flexneri and S. boydii to species
level, for which the serogrouping sera are used.

Serotyping in Shigella is based on O antigen antigenicity only. H antigen typing cannot be developed
since all Shigella strains are defective in flagella expression. Serotyping is a key component of
identification and typing of Shigella. There are currently 55 Shigella serotypes. S. boydii, S.
dysenteriae and S. flexneri have 20, 15 and 19 serotypes, respectively, while S. sonnei has only one
serotype [14,15]. The current 19 S. flexneri serotypes are 1a, 1b, 1c (or 7a), 1d, 2a, 2b, 3a, 3b, 4a, 4av,
4b, 5a, 5b, X, Xv (4c), Y, Yv, 6 and 7b. All S. flexneri serotypes except serotype 6 are minor variants, due
to phage conversion [16] or plasmid-mediated conversion [17], with the majority being due to glucosyl
and/or O-acetyl modifications of the common O units. These modifications are mediated by
glucosyltransferases encoded by serotype-converting bacteriophages, including SfI [17], SfV [18],
SfX [19], SF6 [20] and SfII [21].

It should be noted that Shigella and EIEC share many of the O antigens including O124 (D3), O112
(D2/B15), O143 (B8), O152 (D12), and O167 (B3) [22,23] and thus serotyping cannot be used for
differentiating Shigella from EIEC for these O antigen serotypes. Molecular serotyping methods using an
O antigen-specific gene target from the O antigen gene cluster [24] or restriction polymorphism of the O
antigen gene cluster [25] are available. Most of the S. flexneri serotypes can be typed using a multiplex
PCR based on serotype modification specific genes [26].

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Metabolic regulation by global regulators in response to culture environment

Kazuyuki Shimizu, in Bacterial Cellular Metabolic Systems, 2013

3.6

Acid shock or the effect of pH

The acid barrier of the stomach represents a strong challenge for many pathogenic enterobacteria.
Enteric bacteria that cause disease in the human intestine endure transient but extreme acid conditions
in the stomach. The normal stomach presents an acid environment at around pH 2, with an emptying
time of about 2 h (Smith, 2003). Unlike acid sensitive Vibrio cholerae, E. coli, and Shigella have potent
acid resistant systems able to withstand a low pH at around 2 for at least 2 h (Lin et al., 1995; Castanié-
Cornet et al., 1999). E. coli possesses a level of acid resistance rivaling that of the gastric
pathogen Helicobacter Pylori(Rektorschek et al., 2000). As such, it is important to understand cell
metabolism in relation to acidic conditions, from both medical and fermentation points of view. The
molecular and physiological response to acid stress has thus been the subject of intense investigation
(Foster, 2004; Stincone et al., 2011).

Several acid stress response systems that can protect E. coli from acidic conditions have been
investigated (Foster, 2004; Richard and Foster, 2003, 2004; Stincone et al., 2011). Some of these depend
on the available extracellular amino acids such as glutamate, arginine, and lysine, where the intracellular
proton is consumed by the reductive decarboxylation of the amino acid followed by the excretion of the
product, such as γ-amino butyric acid (GABA) from cytoplasm to the periplasm by a dedicated antiporter
that also imports the original amino acid (Foster, 2004) (Figure 3.26). E. coli cells have been
demonstrated to exhibit acid resistance by such genes as gadAB, which encode glutamate decarboxylase
and gadC, which encode the Glutamate : GABA antiporter. Glutamate decarboxylase production has
been shown to increase in response to acid, osmotic, and stationary phase signals.
The gadA and gadB genes for glutamate decarboxylase isozymes form a glutamate-dependent acid
response system, where the process of decarboxylation consumes an intracellular proton and helps
maintain pH homeostasis. It has also been shown that there exist similar acid resistant systems using
arginine instead of glutamate by arginine decarboxylase, where the antiporter is AdiC in this case (Lin et
al., 1995; Castanié-Cornet et al., 1999; Gong et al., 2003; Iyer et al., 2003), and using lysine by lysine
decarboxylase (Iyer et al., 2003). Note that cells grown in a media rich in amino acids such as LB are acid
resistant (Foster, 2004).

In a typical batch culture, organic acids are most accumulated at the late growth phase or the stationary
phase, and it has been known that GadA and GadB proteins increase in response to the stationary phase
and low pH (Castanié-Cornet and Foster, 2001). The sigma factor σs or RpoS, which increases its amount
at the late growth phase and the stationary phase, as well as Crp, are involved in acid resistance
(Castanié-Cornet et al., 1999; Foster, 2004). As is implied by the involvement of Crp, the resistance
system is repressed when glucose is present. Moreover, it has been shown that FoF1 proton
translocating ATPase is involved in this system (Richard and Foster, 2004). The FoF1 ATPase is utilized as
the protons in the periplasm move into the cytosol across the cell membrane, producing ATP from ADP
and Pi by the negative proton motive force (PMF). Since the basic problem of acid stress is the
accumulated proton in the cytosol, this proton can be pumped out through FoF1 ATPase by hydrolyzing
ATP and reversed proton movement due to positive PMF at a low pH such as pH 2 or 3 (Richard and
Foster, 2004). Without amino acids available in the media, this acid response system is activated by
utilizing FoF1 ATPase (Martin-Galiano et al., 2001; Richard and Foster, 2003), where the positive proton
motive force (PMF) pumps extrudes protons (H+) from the cytoplasm with consumption of ATP (Richard
and Foster, 2004). Namely PMF is operated in the reverse direction, as compared to producing ATP.

Table 3.8 shows 11 regulators involved in regulating glutamate-dependent acid resistance. In the typical
batch culture, the gadA/BC loci can be induced during growth in acidic minimal media (pH 5.5) or in the
stationary phase regardless of pH (Foster, 2004). However, in complex media such as LB, neither locus is
induced until the culture enters into stationary phase.

The expressions of gadA/BC genes are under control of GadE and the response regulator RcsB (Castanié-
Cornet et al., 2010), where RcsB is part of the RcsCDB phosphorelay, a signal transduction system
conserved in members of the Enterobacteriaceae. The RcsB can also be activated independently of the
phosphorelay, by binding of different co-regulators, such as RcsA (main one), RmpA, TviA, and PhoP.
(Castanié-Cornet et al., 2010). As shown in Figure 3.27, EvgS is a sensor kinase and phosphorylates EvgA,
where the phosphorylated EvgA activates the gadE gene as well as ydeO, where YdeO also regulates
the gadE gene. It has also been shown that the small membrane protein B1500 connects the signal
transduction cascade between EvgS/EvgA and PhoQ/PhoP, where b1500 is located upstream
of ydeO and under control of EvgA (Eguchi et al., 2007) and Mg2+ turns off the PhoQ/PhoP system
(Castelli et al., 2000). Moreover, the phosphorylated PhoP activates gadW, and GadW
activates hdeAand gadA, where hdeA is under the control of GadW, PhoP-P, and GadE. EvgA regulates at
least eight genes related to acid resistance, such as ydeP, b1500, ydeO (Matsuda and Church, 2003).

It has been shown that an acid pH lowers cAMP levels in exponentially growing cells in the minimal
glucose medium. This may elevate RpoS that would drive increased expression of gadX. However, GadW
represses RpoS synthesis under acidic conditions, and in turn GadX synthesis. GadX, when not repressed
by GadW, is acid induced due to changes in cAMP. GadW is also acid induced when it is not repressed by
GadX. GadX directly binds to the gadW promoter region. GadX and GadW collaborate to
repress gadA and gadBC expressions under alkaline conditions (Ma et al., 2003). The GadX-GadW
regulon has also been investigated by DNA microarray (Tucker et al., 2003).

It has also been shown that the two-component system of EnvZ (sensor) and OmpR (regulator) regulates
porin expression, where OmpR may be a key regulator for acid adaptation, and thus the opmR mutant is
sensitive to acid exposure (Stincone et al., 2011). It has been shown that the level of OmpC increases
with increased osmolarity when cells are growing in neutral or alkaline media, whereas the level of
OmpF decreases at high osmolarity (Sato et al., 2000). It has also been shown that these porin proteins
play important roles at acidic conditions.

The acid-inducible asr gene is reported to be regulated by the two-component system PhoR/B, which
controls the pho regulon in response to phosphate starvation, and thus the PhoB-PhoR deletion mutant
fails to induce asr gene expression (Suziedeliene et al., 1999). It has also been suggested that H+ directly
or via its acceptor might activate a sensor protein PhoR in the periplasm (Suziedeliene et al., 1999).

Under anaerobic conditions, additional genes, such as ackA and lpdA, as well as hdeA and ompT, are
induced. In order to avoid deleterious concentration in the cell caused by the production by the cell at
low pH, ldhA is induced by acid in order to produce lactate instead of the more harmful acetate plus
formate (Bunch et al., 1997).

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Decarboxylation Test: Types, Uses, principles, procedure and results 4.58/5 (12)

MARCH 16, 2015 BY TANKESHWAR ACHARYAIN BACTERIOLOGY, BIOCHEMICAL TESTS IN

MICROBIOLOGY, LABORATORY DIAGNOSIS OF BACTERIAL DISEASE · 6 COMMENTS

Decarboxylases are a group of substrate specific enzymes that are capable of reacting with the
carboxyl (COOH) portion of amino acids, forming alkaline-reacting amines and byproduct Carbon
dioxide. Increased pH of the medium is detected by color change of the pH indicators bromcresol
purple and cresol red. Bromcresol purple turns purple at an alkaline pH and turns yellow at an acidic pH.

Each decarboxylase enzyme is specific for an amino acid. Lysine, Ornithine, and arginine are the three
amino acids routinely tested in the identification of Enterobacteriaceae.

The specific amine products are:

1. Lysine- Cadaverine

2. Ornithine-Putrescine

3. Arginine- Citrulline

These byproducts are sufficient to raise the pH of the media so that the broth turns purple.

Arginine is hydrolyzed to ornithine ( arginine is first converted to citrulline via dihydrolase reaction, in
which NH2 group is removed from agrinine. Citrulline in next converted to ornithine, which then
undergoes decarboxylation to form putrescine.)

If the inoculated medium is yellow, or if there is no color change, the organism is decarboxylase-
negative for that amino acid. If the medium turns purple, the organism is decarboxylase-positive for that
amino acid.

Requirements

1. Moeller decarboxylase base-4 tubes with lysine, ornithine and agrinine hydrochloride 1% and
control

Method

1. Inoculate the test medium, overlaid with paraffin layer

2. Incubate and read daily for four days

Result
  Purple color-Positive Decarboxylation

 Yellow color-Negative i.e. No decarboxylation

Control

1. Lysine: Klebsiella pneumoniae

2. Ornithine: Enterobacter cloacae

3. Arginine: Enterobacter cloacae

Negative

1. Lysine: Enterobacter cloacae

2. Ornithine: Klebsiella pneumoniae

3. Arginine: Klebsiella pneumonia

Uses;
Lysine Decarboxylase Test (LDC): To assist in the identification of Salmonellae (+ve) and Shigellae (-ve).

Bacteria that are Lysine Decarboxylase (LDC) Positive

1. Escherichia coli

2. Salmonella typhi and Most other salmonellae species (except Salmonella paratyphi A).

3. Klebsiella pneumoniae

4. Serratia marcescens

5. Vibrio cholerae

6. Vibrio Parahemolyticus