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Abstract: Ripening of fleshy fruit is a differentiation process involving biochemical and biophysical changes that lead
to the accumulation of sugars and subsequent changes in tissue texture. Also affected are phenolic compounds, which
confer color, flavor/aroma, and resistance to pathogen invasion and adverse environmental conditions. These phenolic
compounds, which are the products of branches of the phenylpropanoid pathway, appear to be closely linked to fruit
ripening processes. Three key enzymes of the phenylpropanoid pathway, namely phenylalanine ammonia lyase, Omethyltransferase, and cinnamyl alcohol dehydrogenase (CAD) have been reported to modulate various end products
including lignin and protect plants against adverse conditions. In addition, peroxidase, the enzyme following CAD
in the phenylpropanoid pathway, has also been associated with injury, wound repair, and disease resistance. However,
the role of these enzymes in fruit ripening is a matter of only recent investigation and information is lacking on the
relationships between phenylpropanoid metabolism and fruit ripening processes. Understanding the role of these enzymes
in fruit ripening and their manipulation may possibly be valuable for delineating the regulatory network that controls the
expression of ripening genes in fruit. This review elucidates the functional characterization of these key phenylpropanoid
biosynthetic enzymes/genes during fruit ripening processes.
Introduction
Fruit ripening is a developmentally regulated process resulting
from the coordination of numerous biochemical and physiological
changes within the fruit tissue that culminates in changes in fruit
firmness, color, taste, aroma, and texture of fruit flesh (Figure 1)
(Brummell and Harpster 2001; Vicente and others 2006; Singh
and others 2007). Textural changes that lead to softening of fruits
have drawn attention to the putative involvement of several enzymes able to act and modify its structure in a developmental and
coordinated way (Brady 1987; Rose and others 1998; Sozzi and
others 1998).
Phenolic compounds produced by the phenylpropanoid pathway contribute to fruit pigmentation and the disease resistance
response found in many fleshy fruits during ripening (Figure 2).
In olive fruits, several simple and complex phenolics have been
reported to be effective against pathogenic bacteria (Chowdhury
and others 1997). Caffeic acid has been found to be the most effective agent, although oleuropein, the major phenolic constituent
of olives, also exhibits bactericidal action (Ruiz-Barba and others
1991). Moreover, phenolic compounds protect plants by acting
as feeding deterrents to insects (Nahrstedt 1990). For example,
flavonoids such as luteolin, naringenin, phloretin, quercetin 3rhamnoside, and myricetin-3-rhamnoside, have been shown to
act as feeding deterrents to aphids (Dreyer and Jones 1981).
In addition, a complex polymer, lignin, is also a product of phenylpropanoid metabolism. Lignin represents a major carbon sink in vascular plants and is derived from 3
lignin precursors termed as monolignols (hydroxycinnamyl
alcohols), namely, p-coumaryl alcohol, coniferyl alcohol, and
sinapyl alcohol. The biosynthesis of monolignols involves 2 specific steps branching off the general phenylpropanoid pathway.
The phenylpropanoid pathway drives the carbon flow from
the aromatic amino acid L-phenylalanine (L-Phe) or, in some
cases, L-tyrosine (L-Tyr) (Rosler and others 1997), for the
production of 4-coumaroyl CoA (or a respective thiol ester
in the presence of other 4-hydroxycinnamates). The synthesis of monolignols (lignin monomers) involves several hydroxylation/methylation and oxidation/reduction reactions (Whetten
and Sederoff 1995) (Figure 3). Numerous enzymes, namely, a
family of oxygen-dependent cytochrome P450 hydroxylases and
S-adenosylmethionine-dependent methyltransferases, act on the
free cinnamic acids and their CoA esters which are converted to
p-coumaryl alcohol, coniferyl alcohol, and sinapyl alcohol. After
synthesis in cytoplasm, monolignols are transported to the cell wall
as glucoside derivatives formed in a reaction catalyzed by uridine
diphosphate glucose coniferyl alcohol glucosyltransferase (UDPGT), where these glucoside derivatives are hydrolyzed by coniferin
-glucosidase (CBG). The last major step in lignin biosynthesis
is monolignol dehydrogenation in a reaction catalyzed by peroxidase (POD), laccase (LAC), polyphenol oxidase (PO), or coniferyl
alcohol oxidase (CAO) followed by polymerization by oxidative
coupling (Boerjan and others 2003). Peroxidases are involved in
the oxidation of phenolic compounds in cell walls, polymerization of lignin and suberin, and have also been shown to degrade
flavonoids in noncell systems (processed food) as well as in vivo. As
lignin polymerizes, it serves as a matrix around the polysaccharide
398 Comprehensive Reviews in Food Science and Food Safety r Vol. 9, 2010
c 2010 Institute of Food Technologists
doi 10.1111/j.1541-4337.2010.00116.x
S
H
I
K
I
M
A
T
E
Phosphoenolpyruvate
Glycolysis
P
A
T
H
W
A
Y
Phenylalanine
P
H
E
N
Y
L
P
R
O
P
A
N
O
I
D
P
A
T
H
W
A
Y
F
L
A
V
O
N
O
I
D
P
A
T
H
W
A
Y
Benzoic acid
derivatives
Cinnamate
Esters of
organic acids
4-coumarate
MONOLIGNOLS
(p-coumaryl alcohol, coniferyl alcohol, sinapyl alcohol)
malonylCoA
Lignin
(Biopolymer of aromatic subunits
forming an integral part of the
secondary cell walls of plants)
Carbohydrate
metabolism
Chalcone
(Aromatic ketone that forms the central core for a variety of important
biological compounds)
Flavones
[Flavonoids based on the backbone of 2-phenylchromen-4-one
(2-phenyl-1-benzopyran-4-one)]
Leucoanthocyanidins
(Chemically flavan-3,4-diols which are colorless compounds
related to anthocyanidins and anthocyanins)
Anthocyanins
(Water-soluble vacuolar pigments which are glucosides of
anthocyanidins and may appear red, purple or blue according to pH)
Proanthocyanidins
(Polymers of flavonoids)
Figure 1Schematic overview of branch pathways of shikimate, phenylpropanoid metabolism, and flavonoid biosynthesis in plants leading to the
synthesis of flavonoids.
c 2010 Institute of Food Technologists
Vol. 9, 2010 r Comprehensive Reviews in Food Science and Food Safety 399
Caffeic acid
(Source of O-diphenols
oxidizable, fungitoxic)
Zwitterionic anthocyanin
(Protectant)
OH
C
OH
C
OH
C
OH
OH
OH
OH
C
NH2
PAL
C4H
HCH
OMT
F5H
OH
OH
OCH3 HO
OH
OMT
OH
Ferulic Acid
Esterification of host
cell wall
(Chemical barrier)
OCH3 H3CO
OH
OCH3
OH
5 OH-Ferulic Acid
Sinapic Acid
Stress lignin
(Physical Barrier to fungus)
Figure 2The involvement of phenolic compounds in the expression of resistance to pathogen infection. The diagram shows the family of compounds
known as phenylpropanoids and their route of synthesis from phenylalanine through the phenylpropanoid pathway. PAL-phenylalanine ammonia
lyase, C4H-cinnamic acid 4-hydroxylase, HCH-hydroxycinnamate 3-hydroxylase, OMT-O-methyl transferase, F5H-ferulic acid-5-hydroxylase.
tissue) and in the vascular bundles that connect the achenes to the
central pith (Suutarinen and others 1998).
Caffeoyl CoA OMT (CCoAOMT) is induced in elicited cells
synthesizing phenylpropanoid phytoalexins and, therefore, has
been implicated in the defense response of plant cells (Schmitt
and others 1991). In another study, the analysis of the effect of
high CO2 treatment on phenolic metabolism and ripening-related
changes in cherimoya fruit (Annona cherimola) demonstrated no
change in the total polyphenol levels. Upon exposure to air, however, there was a slight decrease in lignin content in CO2 -treated
fruits but the levels remained significantly higher compared to airtreated controls (Maldonado and others 2002). The data indicated
that high CO2 treatment promoted changes in lignin degradation. It was hypothesized that cell walls maintaining more lignin
deposits could modulate the strength of cellcell adhesion (Assis
and others 2001). Furthermore, in considering the insecticidal effects attributed to enriched CO2 treatment and the importance of
phenylpropanoid compounds in general defense strategies, it has
been suggested that the maintenance of these compounds in CO2 treated fruit may be an advantage in pathogen defense (Assis and
400 Comprehensive Reviews in Food Science and Food Safety r Vol. 9, 2010
c 2010 Institute of Food Technologists
OH
C
NH2
TAL
O
OH
OH
OH
S-CoA
COAMP
OH
NH2
PAL
B
4CL
C4H
C
4CL
CCR
D
OH
OH
CAD
OH
OH
C3H
G
OH
CCoA3H
OH
COAMP
4CL
4CL
OH
OH
S-CoA
OH
OH
OH
OH
CCoAOMT
OMT
OH
COAMP
4CL
4CL
OCH3
OCH3
OH
OH
CCR
CAD
OCH3
OH
CHO
SCoA
OCH3
OH
OH
OCH3
OH
F5H
OH
COAMP
HO
OCH3
HO
OCH3 HO
OH
OH
OCH3
P
OH
CCoAOMT
OH
COAMP
OCH3 H3CO
OH
SCoA
4CL
4CL
H3CO
OCH3 H3CO
OH
OMT
O
CCR
4CL
4CL
Lignin and
Lignans
Synthesis
CHO
SCoA
OCH3 H3CO
OH
CCR
R
OH
OCH3 H3CO
OH
CHO
CAD
S
OH
OCH3 H3CO
OCH3
OH
Figure 3An overview of the monolignol biosynthetic pathway (A) tyrosine, (B) L-phenylalanine, (C) cinnamic acid, (D) p-coumaric acid, (E)
p-coumaroyl CoA, (F) p-coumaraldehyde, (G) p-coumaryl alcohol, (H) caffeic acid, (I) caffeoyl CoA, (J) ferulic acid, (K) feruloyl CoA, (L) coniferaldehyde,
(M) coniferyl alcohol, (N) 5-hydroxyferulic acid, (O) 5-hydroxyferuloyl CoA, (P) 5-hydroxyconiferaldehyde, (Q) sinapic acid, (R) sinapoyl CoA, (S)
sinapaldehyde, (T) sinapyl alcohol. TAL, tyrosine ammonia lyase; PAL, phenylalanine ammonia lyase; C4H, cinnamate 4-hydroxylase; 4CL, 4-coumarate
CoA ligase; CCR, cinnamoyl CoA reductase; CCoAOMT, caffeoyl CoA 3-O-methyltransferase; F5H, ferulate 5-hydroxylase; ?, yet unconfirmed reactions.
c 2010 Institute of Food Technologists
Vol. 9, 2010 r Comprehensive Reviews in Food Science and Food Safety 401
402 Comprehensive Reviews in Food Science and Food Safety r Vol. 9, 2010
c 2010 Institute of Food Technologists
Serial nr
Plant
description
Accession
nr
1.
Vitis vinifera
EF192469
2.
Prunus avium
AF036948
3.
Musa acuminata
EU104680
4.
Eriobotrya japonica
EF685344
5.
Malus X domestica
AF494403
6.
Pyrus communis
DQ230992
7.
Citrus limon
U43338
8.
Fragaria X ananassa
AB360390
9.
Rubus idaeus
AF237955
10.
Ipomoea batatas
M29232
11.
Cucumis melo
X76130
12.
Lycopersicon esculentum
M83314
Maximum
identity
with fruits
Maximum
identity
other plants
Enzyme
activity
Prunus avium
AF036948 (80%)
Pyrus communis
DQ230992 (88%)
Camellia sinensis
D26596 (79%)
Robinia pseudoacacia
EU650628 (80%)
Citrus clementina X
Citrus reticulate
AJ238754 (83%)
Pyrus communis
DQ901399 (95%)
Pyrus communis
DQ230992 (97%)
Prunus avium
AF036948 (88%)
Populus trichocarpa
EU603319 (84%)
Quercus suber
AY443341 (80%)
Populus euramericana
AJ698920 (79%)
Populus tomentosa
EU760386 (78%)
C. clementina X C.
reticulate
AJ238753 (77%)
Rubus idaeus
AF237955 (92%)
Prunus avium
AF036948 (83%)
Lycopersicon
esculentum
M83314 (80%)
Populus trichocarpa
EU603320 (81%)
Lotus japonicas
AB283033 (79%)
Camellia sinensis
D26596 (78%)
Nicotiana tabacum
(Samsun NN)
X78269 (79%)
Trifolium pretense
DQ073811 (78%)
S. tuberosum X63103
(92%)
Ipomoea nil
AF325496 (80%)
c 2010 Institute of Food Technologists
mRNA
accumulation
the other in nearly ripe fruit (Cheng and Breen 1991). The 1st
peak was suggested to be involved in the synthesis of flavonoids
(condensed tannins) and phenolics that took place during early
fruit development, whereas the 2nd activity peak was associated
with the anthocyanin accumulation that occurred during later
stages of fruit ripening (Macheix and others 1990). In orange
fruit, the expression of a putative anthocyanin transporting glutathione S-transferase (GST) was correlated with the expression of
the pal, chalcone synthase (chs), dihydroflavonol 4-reductase (dfr),
and UDP glucose, flavonol 3-O-glucosyltransferase (ufgt) genes
under cold stress (Lo Piero and others 2005, 2006). As is the case
for many secondary metabolite biosynthetic proteins, an apparent redundancy with the anthocyanin-transporting GSTs in grape
berries (Vitis vinifera) was reported, and this redundancy was seen
with numerous functional copies of biosynthetic enzymes including PAL (Conn and others 2008). It is quite clear that, even for
much-studied old pathways like flavonoid biosynthesis, these
are exciting times.
In an earlier study, expression of 7 genes of the anthocyanin
biosynthetic pathway (pal, chs, chi, f3h, dfr, ldox, and ufct) was
determined in Shiraz grape berries (V. vinifera). In flowers and
grape berry peels, expression of all of the genes, except ufct, was
detected up to 4 wk postflowering, followed by a reduction in this
expression 6 to 8 wk postflowering. Expression of chs, chi, f3h, dfr,
ldox, and ufct then increased 10 wk postflowering, coinciding with
the onset of anthocyanin synthesis. The results obtained in the
study provide additional evidence for the correlation between the
expression of structural flavonoid pathway genes and anthocyanin
production during fruit development. In grape berry flesh, no pal
or ufct expression was detected at any stage of development, but
chs, chi, f3h, dfr, and ldox were expressed up to 4 wk postflowering.
These results indicated that the onset of anthocyanin synthesis in
ripening grape berry peel coincides with a coordinated increase in
Vol. 9, 2010 r Comprehensive Reviews in Food Science and Food Safety 403
O-Methyltransferases (OMTs)
Methyltransferases are ubiquitous enzymes that catalyze the
transfer of a methyl group from S-adenosyl-L-methionine (SAM)
to an acceptor substrate, generating O-, N-, S-, and C-methyl
derivatives and S-adenosyl homocysteine (Ibrahim and others
1998). SAM-dependent O-methylation is catalyzed by OMTs
and involves the transfer of the methyl group of S-adenosylL-methionine (AdoMet) to the hydroxyl group of an acceptor
molecule, with the formation of its methyl ether derivative and
S-adenosyl-L-homocysteine as products such as lignin, flavonoids,
phenylpropanoids, and alkaloids (Dwivedi and others 1994; Pichersky and Gang 2000; Rastogi and Dwivedi 2006). OMTs play
a critical role in the biosynthesis of many classes of compounds
required for plant growth, aroma generation, and plant defense.
There are several hundred O-methylated flavonoids which occur in
plants, and these range from mono- to polymethylated compounds
belonging to the chalcones, flavones, isoflavones, and flavonols, as
well as their dihydro derivatives (Wollenweber and Dietz 1981).
Combined biochemical and molecular analyses of volatile components produced by fruit have demonstrated that their biogenesis
forms an integral part of ripening. Several omt cDNA clones have
been reported from different plant species, which share common
structural as well as physicochemical features. The phylogenetic
analysis of plant omt sequences suggests that plant omts may have
diverged from a common ancestral gene, through gene duplication
404 Comprehensive Reviews in Food Science and Food Safety r Vol. 9, 2010
c 2010 Institute of Food Technologists
COMT
COMT
OH
OCH3 HO
OH
OH
OCH3
H3CO
OCH3
OH
OH
R-COOH:Caffeic Acid
R-COOH:Ferulic Acid
R-COOH:5-Hydroxyferulic Acid
R-COOH:Sinapic Acid
R-CHO:Caffeoyl Aldehyde R-CHO:Coniferyl Aldehyde R-CHO:5-Hydroxyconiferyl Aldehyde
R-CHO:Sinapyl Aldehyde
R-COH:Caffeoyl Alcohol
R-COH:Coniferyl Alcohol R-COH:5-Hydroxyconiferyl Alcohol R-COH:Sinapyl Alcohol
COOH
OH
OMT
OH
OH
OH
OCH3
Catechol
OCH3
Guaiacol
OH
Ferulic Acid
[A]
OH
OH
OMT
OCH3
OMT
OH
OH
Caffeic Acid
COOH
CHO
Protocatechuic
Aldehyde
[B]
CHO
Vanillin
[C]
CCoAOMT (EC 2.1.1.104), which catalyzes the meta-Omethylation of caffeoyl CoA to form feruloyl CoA, appears to
play an important role in the formation of the coniferyl alcohol
moieties that are precursors for lignification (Dwivedi and Campbell 1995; Vander and others 2000). CCoAOMT catalyzes 1 of 2
alternative methylation steps of the phenylpropanoid biosynthesis
pathway which leads to the synthesis of diverse secondary products such as lignin, flavonoids, and isoflavonoids (Ye and Varner
Classification of plant OMT
OMTs are widespread throughout the plant kingdom and found 1995) and this methylation step is highly regulated (Hahlbrock and
in all lignin-producing plants. O-methylation of the phenyl- Scheel 1989).
propanoid and flavonoid groups of compounds is catalyzed by
distinct classes of OMTs.
Proposed methyl acceptor classes and groups
The molecular and biochemical data available so far provide the
Caffeic acid 3-O-methyltransferase (COMT, EC 2.1.1.68) catalyzes the O-methylation of aromatic diols and is involved in basis for a meaningful classification of the plant omt gene superfamlignification (Rastogi and Dwivedi 2008) or may have other ily. There exist some 36 omt cDNA clones, which are subdivided
physiological functions like flavor generation (Collendavelloo and into 5 groups encoding the methylation of the lignin precursors,
others 1981; Pellegrini and others 1993) (Figure 4). Interest- caffeic and 5-hydroxyferulic acids (Group 1), flavonoids (Group 2),
ingly gene evolution studies in Clarkia breweri have suggested that and phenylpropanoids (such as the coumarins Group 3). Group 4 is
isoeugenol OMT (iemt) gene that catalyzes the methylation of primarily comprised of simple phenols and anthocyanins, whereas
eugenol and isoeugenol to form the volatiles methyleugenol and Group 5 encompasses polyketides and other acetate/malonateisomethyleugenol had arisen from comt gene. It was suggested that derived compounds (Table 2). Each group of OMT is further
OMT substrate preference could be regulated by a few amino classified into Class A and Class B. Based on the class of subacid residues, and new OMTs with different substrate specifici- strate methylation, Class A OMTs methylate phenylpropanoid
ties could evolve from an existing OMT by mutation of several compounds, whereas Class B OMTs methylate flavonoid comamino acids (Wang and Pichersky 1999). The evolution of new pounds. Although the chemical mechanisms of methyl transfer
OMTs with new substrate specificities is relatively simple; it is not reactions are identical, OMTs differ in their selectivity with resurprising that OMTs with similar substrate specificities evolved spect to the stereochemistry of the methyl acceptor molecules, as
independently in different plant lineages more than once. These well as the substitution pattern of their phenolic hydroxyl groups
broad-specificity enzymes may be recruited for new metabolic (Ibrahim and others 1998). Some of the known OMTs display
pathways, followed by further evolution toward more specific and strict specificities toward their acceptor substrate as well as to the
efficient catalysts. It follows that gene duplication (or even poly- position of substrate methylation. In contrast, other OMTs, espeploidy) is an important factor in this concept as it provides the cially those catalyzing the methylation of catechol (O-dihydroxy)
raw material for the acquisition of new biosynthetic pathways. It moiety substrates, exhibit surprisingly broad substrate specificities.
is somewhat surprising that developing strawberry fruits display OMTs have been shown to be multifunctional enzymes that could
high OMT activity levels toward caffeic acid, protocatechuic alde- also catalyze transformations in 2 different biosynthetic pathways
hyde, and catechol resulting in ferulic acid, guaiacol, and vanillin such as the alkaloid and phenylpropanoid pathways (Li and others 1997) or aroma biosynthesis (Lavid and others 2002) and,
(Figure 5A to 5C), respectively.
and mutation, to yield the various functional enzyme groups currently recognized in plants (Ibrahim 1997). It would be interesting
to group plant omt cDNA genes according to a functional trait
that reflects the substrate preferences of their encoded proteins,
and could be used as the basis for classification of this supergene
family.
c 2010 Institute of Food Technologists
Vol. 9, 2010 r Comprehensive Reviews in Food Science and Food Safety 405
Groups
Group 1
Group 2
Group 3
Group 4
Group 5
406 Comprehensive Reviews in Food Science and Food Safety r Vol. 9, 2010
c 2010 Institute of Food Technologists
OH
C4H
p-Coumaric acid
HOOC
PAL
HOOC
NH3 Phenylalanine
HSAOC
OH
HO
COOH
H2C
OH
STS
HO
Trihydroxychalcone
Tetrahydroxychalcone
HO
Stilbene
F3'H
Nf F3'5'H
HO
HO
OH
UFGT
Delphinidin
O-Glc
OH
IOMT
OH
Flavonol glycosides
R
OCH3
I2'H
OH
HO
OH
Petunidin-3-glucoside
O
O
OH
Guc
OMT
HO
OCH3
HO
OCH3
IFR
Delphinidin-3-glucoside
O
OH
OH
OH
HO
RT
OCH3
OMT
HO
Isoflavone
R'
O-Glc-O-Rha
OH
OH
OH
OH
UFGT
R'
OH
HO
OH
Cyanidin
Flavonols
FSLI
OH
IFS
OH
LAR
Catechins
Catechins LAR Leucocyanidin Flavonols Leucodelphinidin
OH
LDOX
LDOX OH BAN
BAN
OH
OH Rha-O
O
H
R
HO
Phf
Dhk
DFR
OH
Aurones
F3H
F3'5'H Dhm
DFR
FLSI
F3H
Flavanone
OH
F3H
OH
O
CH
CHI
CHI
O
OH
HO
OH
OH
OH
OH
Naringenin Chalcone
HO
OH
OH
OH
OH
COSCoA
MalonylCoA
4-Coumaroyl-CoA
CHS
CHS
HO
CHS/CHR
OCH3
Guc
OMT
OH
Cyanidin-3-glucoside
OCH3
OH
HO
OCH3
Malvidin-3-glucoside
VR
DMID
Peonidin-3-glucoside
2'-hydroxy Isoflavanone
HO
OCH3
Isoflavanoids
Figure 6Metabolic pathway and key steps of the flavonoid biosynthesis pathway leading to anthocyanin formation. The figure also depicts the
cross-linking and interdependence of flavonoid and lignin biosynthesis pathways. Acronyms of the compounds reported in the figure stand for the
following: 4CL, 4-coumaroyl CoA ligase; I2 H, isoflavone 2 -hydroxylase; BAN, anthocyanidin reductase; C4H, cinnamate-4-hydroxylase; CHI, chalcone
isomerase; CHR, chalcone reductase; CHS, chalcone synthase; DFR, dihydroflavonol 4-reductase; Dhk, dihydrokaempferol; Dhm, dihydromyricetin; Dhq,
dihydroquercetin; DMID, 7,2 -dihydroxy, 4 -methoxyisoflavonol dehydratase; E, eriodictyol; F3 5 H, flavonoid 3 ,5 -hydroxylase; F3 H, flavonoid
3 -hydroxylase; F3H, flavanone 3-hydroxylase; FLS1, flavonol synthase; IFR, isoflavone reductase; IFS, isoflavone synthase; IOMT, isoflavone
O-methyltransferase; LAR, leucoanthocyanidin; LDOX, leucoanthocyanidin dioxygenase; Nf, naringenin flavanone; PAL, phenylalanine ammonia lyase;
Phf, pentahydroxyflavanone; RT, rhamnosyl transferase; STS, stilbene synthase; UFGT UDP, glucose, flavonol 3-O-glucosyltransferase; VR, vestitone
reductase.
c 2010 Institute of Food Technologists
Vol. 9, 2010 r Comprehensive Reviews in Food Science and Food Safety 407
HO
OH
and terpene volatile compounds (Dandekar and others 2004). Interestingly, due to the huge number of flavor compounds found in
strawberry, Zabetakis and Holden (1997) suggested that it was not
possible for each substance to have its own enzymes. Accordingly, the side activity of COMT involved in phenylpropanoid
metabolism of strawberry fruits was thought to be responsible
for the formation of 2,5-dimethyl-4-methoxy-3(2H)-furanone
(DMMF). Consistent with this view, it was reported that a single OMT from Chrysosplenium americanum could methylate both
flavonoid and phenylpropanoid compounds (Gauthier and others
1998). Of the 15 volatiles in strawberry, 4-hydroxy-2,5-dimethyl3(2H)-furanone (HDMF) was regarded as vital, but it was methylated further by FaOMT (Fragaria X ananassa OMT) to DMMF
during the ripening process (Wein and others 2001) (Figure 7A).
HDMF was indispensable because of its high concentration (up
to 55 mg kg1 strawberry fruit FW) (Larsen and others 1992)
and low odor threshold (10 ppb in water) (Schieberle and Hofmann 1997). The reduction of faomt gene expression altered the
HDMF/DMMF ratio, resulting in a near-depletion of the DMMF
pool, thus confirming the importance of FaOMT in the DMMF
formation. faomt encoded a sequence of 365 amino acids and accepted a substrate spectrum for compounds containing O-diphenol
structures (Wein and others 2002).
In addition, the dual function of this enzyme in the secondary
metabolism was proved as faomt down-regulation and also affected
the concentration of feruloyl 1-O--d-glucose and caffeoyl 1-O-d-glucose, suggesting that it was also involved in the methylation
of the caffeoyl group (Lunkenbein and others 2006) (Figure 7B).
HO
OH
OCH3
HDMF
O
FaOMT
OH
DMMF
[A]
HO
COOH
H3CO
COOH
FaOMT
HO
Caffeic Acid
Ferulic Acid
O
OH
HO
O
HO
HO
O
HO
Caffeoyl -D-glucose
H3CO
OH OH
OH
O
HO OH
HO
OH
Feruloyl -D-glucose
[B]
408 Comprehensive Reviews in Food Science and Food Safety r Vol. 9, 2010
c 2010 Institute of Food Technologists
involving the accumulation of lignin, flavonoids, and polysaccharides which was thought to be due to cad induction (Kpemoua
and others 1996). CAD is therefore regulated by both developmental and environmental stimuli, much like other well-studied
enzymes of phenylpropanoid metabolism (Whetten and Sederoff
1995).
Vol. 9, 2010 r Comprehensive Reviews in Food Science and Food Safety 409
410 Comprehensive Reviews in Food Science and Food Safety r Vol. 9, 2010
c 2010 Institute of Food Technologists
Chalcone Synthase
Chalcone synthase (CHS, EC 2.3.1.74) is the enzyme responsible for catalyzing the 1st committed step of the flavonoid biosynthesis pathway. CHS is an acyltransferase enzyme that catalyzes
the condensation of 4-coumaroyl CoA to the 1st flavonoid naringenin chalcone, in the presence of 3 molecules of malonyl CoA
c 2010 Institute of Food Technologists
Peroxidases
Peroxidase (POD, EC 1.11.1.7) catalyzes the polymerization of
phenylpropanoid precursors of lignin and are involved in the last
step of lignin formation. However, it has been difficult to differentiate between peroxidase isozymes associated with lignification
and those involved in biochemical activities (Whetten and Sederoff
1995). Ryugo (1964) followed the changes in lignin and phenolic
precursors in the developing peach pit and suggested it was a good
system for lignin synthesis studies. In peach, a 2-fold increase in
total peroxidase activity during lignifications was reported which
was established by an increase in a number of basic isozymes in
lignifying tissues. Although an increase in the amount and diversity of basic peroxidases was observed during lignification, other
enzymes involved in producing phenylpropanoid precursors may
play a more important role in controlling lignin formation (Abeles
and Biles 1991). Dalet and Cornu (1988) reported no differences in
the amount or distribution of peroxidase isozymes in cherry clones
with different degrees of lignifications. Interestingly, the ejpod gene
cloned from E. japonica exhibited only 1 of 6 amino acid residues
Vol. 9, 2010 r Comprehensive Reviews in Food Science and Food Safety 411
Acknowledgments
The financial assistance from the Dept. of Biotechnology
(DBT), New Delhi, India (in the form of DBT-SRF to Rupinder
Singh), is gratefully acknowledged. We are also thankful to DSTFIST, CSIR (NMITLI), and UP Government (Under Centre of
Excellence in Biochemistry and Biotechnology) for their financial
support in the form of infrastructural facilities.
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