12158 J. Phys. Chem.

B 2008, 112, 12158–12167

Cis-Trans Isomerizations of β-Carotene and Lycopene: A Theoretical Study

Wen-Hsin Guo, Cheng-Yi Tu, and Ching-Han Hu*

Department of Chemistry, National Changhua UniVersity of Education, Changhua 50058, Taiwan
ReceiVed: March 5, 2008; ReVised Manuscript ReceiVed: June 24, 2008

The all-trans to mono-cis isomerizations of polyenes and two C40H56 carotenes, β-carotene and lycopene,
at the ground singlet (S0) and triplet (T1) states are studied by means of quantum chemistry computations. At
the S0 state of polyenes containing n acetylene units (Pn), we find that the energy barrier of the central CdC
rotation decreases with n. In contrast, however, at the T1 state, the rotational barrier increases with n. For the
C40H56 carotenes, the rotational barriers of lycopene are lower than those of their β-carotene counterparts.
This difference renders the rotational rates of lycopene to be 1-2 orders of magnitude higher than those of
β-carotene at room temperature. For both these carotenes, the barrier is lowest for the rotation toward the
13-cis isomer. The relative abundances are in the following order: all-trans > 9-cis > 13-cis > 15-cis. Although
the 5-cis isomer of lycopene has the lowest energy among the cis isomers, its formation from the all-trans
form is restricted, owing to a very large rotational barrier. The possible physiological implications of this
study are discussed.

1. Introduction spinach.18 Further, 9-cis-R-carotene is bound to the Cyt b6f

Carotenes play an important role in biological systems. They
act as antennas in the light-harvesting units in chlorophyll and
bacteriochlorophyll, and they also protect biological tissues from
the deleterious effects of singlet oxygen and triplet bacteriochlo-
rophyll.1,2 Dietary carotenes are known to exhibit antioxidation
properties.3,4 For example, Di Mascio et al. have shown that
lycopene (Scheme 1) is most effective among carotenes in
quenching singlet oxygen.5 Some carotenes, such as β-carotene
(Scheme 1), are converted to retinol (vitamin A); therefore, they
thus are vital for vision, skin and membrane protection, and
anticancer activity.6
The protective effects of carotenoids have been well-documen-
ted.3,4,7-11 It has been recognized by many studies that these
effects are dependent upon their stereochemistry. For example,
the provitamin A activity of carotene is correlated with its
stereogeometry. In living organisms, the conversion of β-car-
otene to retinol is performed via regiospecific, central cleavage
at its (E)-configured 15,15′-position.12 The isomerization of
β-carotene would probably decrease the provitamin A activity.
Although the all-trans isomers of carotenes are apparently
the most stable, cis isomers were found to be present in minor
amounts. In human serum and tissues, it has been discovered
that more than 50% of lycopene and ∼5% of β-carotene exist
in cis conformations.13 The cis isomers of lycopene were found
to be more bioavailable than the all-trans isomer.14 Food
processing, heating, and illumination would definitely facilitate
the cis-trans isomerizations of carotenoids and thus would alter
the bioactivity of these compounds.15,16
In an extract of the thylakoid membranes of a thermophilic
blue-green alga and of spinach, Ashikawa et al. found ap-
proximately 80% all-trans isomer and 20% cis isomers including
9-cis, 9,13-di-cis, 9,9′-di-cis, 13-cis, and 15-cis isomers (in
decreasing order of quantity).17 Different types of carotenes were
observed in the cytochrome b6f (Cyt b6f) complexes obtained
from various species. For one example, 9-cis-β-carotene was
found to be the predominant component in Cyt b6f complex of
* Corresponding author.
10.1021/jp8019705 CCC: $40.75  2008 American Chemical Society
Published on Web 08/28/2008
complex of green alga Bryopsis corticulans.19 Interestingly, it
was observed that in the reaction center of light-harvesting
system of photosynthetic bacteria, the configuration of the
carotenoid is 15-cis; in contrast, carotenoid in the antenna
complexes exhibit all-trans configuration.1 An important ques-
tion has been raised by Ashikawa et al. with regard to whether
or why different isomers in the photosystem have optimal
functions, i.e., photoprotection in the reaction center and light-
harvesting in the antenna complex.1
Because of the relevance to their functions, the isomerization
of carotenoids has attracted considerable attention from chemists.
Spectroscopic identifications of various cis and trans isomers
of β-carotene have been reported by Koyama’s group; in their
study, isomerizations at the S0 and T1 states were investigated
under thermal and triplet-sensitized conditions.20 Doering and
co-workers reported cis-trans thermal isomerizations among
β-carotene and its cis isomers.21 At and above physiological
temperature they found that 13-cis and 15-cis isomers are
relatively abundant, and this leads to the inference that the 13-
cis and 15-cis isomers of β-carotene are true anticarcinogenic
agents, whereas all-trans-β-carotene is relegated to the reservoir
of the cis isomers.21 Doering’s research brought to attention
9-cis, 13-cis, and 15-cis isomers in addition to all-trans-β-
carotene.
After photoexcitation to its optically allowed singlet state,
carotene decays to the lower singlet (S1) and triplet (T1) states
within 1 ps. The lifetimes of S1 and T1 states are in the
picosecond and nanosecond ranges in light-harvesting systems,
respectively.20 In solution, the triplet lifetime of a carotene is
even longer, being at the microsecond range.22 In the presence
of light, therefore, it is thus essential to consider the isomer-
ization process that occurs via the T1 state. In the present
investigation, the cis and trans isomers of β-carotene and
lycopene, and the transitions between them at the S0 and T1
states, are studied using theoretical methods.
All-trans polyenes have been used as model compounds for
understanding the cis-trans isomerizations of carotenes, through
both experimental and theoretical means.21,23-25 Doering and
Cis-Trans Isomerization of β-Carotene and Lycopene J. Phys. Chem. B, Vol. 112, No. 38, 2008 12159

SCHEME 1: Structure and Numbering Schemes for all-trans-β-Carotene and all-trans-Lycopene

TABLE 1: Barriers of Activation Energy (∆Eq), Barriers of

coauthors measured the enthalpies of activation of semirigid Activation Enthalpy (∆Hq), Vertical and Adiabatic
polyenes; they incorporated all the double bonds, but the central Singlet-Triplet Energy Separations (∆E′S-T and ∆ES-T) of
All-Trans Polyenes (in kcal/mol) Predicted by DFT and ab
one, into fused six-membered rings to preclude the formation
Initio Theoriesa
of conformers other than central-cis isomers. Polyenes with up
to nine acetylene units (P9) have been investigated, and the ∆Eq ∆Hq ∆E′S-T ∆ES-T
enthalpies of activation were reported by Doering’s group.24 In P1
this study, we include the central-cis isomerizations of all-trans B3LYP/6-31G(d) 57.4 (58.4)b 57.7 104.1 60.4
polyenes up to P15. CASMP2/6-311G(d)//CAS(2,2)/ 64.4 65.4 110.3 64.1
6-31G(d)
2. Computational Approach CASPT2/6-31G(d)//B3LYP/ 58.3 59.6 104.5
6-31G(d)
The computations were performed using the density functional P2
theory (DFT) and CASMP2 approaches implemented in the B3LYP/6-31G(d) 48.7 (49.3)b 48.8 75.4 54.5
Gaussian03 series of programs.26 In the CASMP227 approach, CASMP2/6-311G(d)//CAS(4,4)/ 55.7 55.8 81.4 59.9
electron correlation corrections are added to the CASSCF 6-31G(d)
theory,28 and it is used in our model polyene compounds. The CASPT2/6-31G(d)//B3LYP/ 50.3 50.0 74.5
B3LYP density functional was applied to all of the compounds 6-31G(d)
in this study; it is a hybrid method that includes Becke’s three- P3
parameter mixing of the nonlocal exchange potential29 and the B3LYP/6-31G(d) 40.5 (41.0)b 40.4 58.6 41.3
nonlocal correlation functional proposed by Lee, Yang, and CASMP2/6-311G(d)//CAS(6,6)/ 45.8 45.6 61.9 44.3
6-31G(d)
Parr.30 In the study by Bernardi et al., it has been demonstrated
CASPT2/6-31G(d)//B3LYP/ 40.7 40.4 58.6
the B3LYP/6-31G(d) theory is able to obtain rotational barriers 6-31G(d)
of small polyenes (up to all-trans-octatetraene) that are within
P4
1 kcal/mol from the CASPT2 predictions.31,32 Harmonic vibra-
B3LYP/6-31G(d) 36.4 (36.7)b 36.2 48.6 32.9
tional frequencies were computed via analytic energy second CASMP2/6-311G(d)//CAS(8,8)/ 40.6 40.8 53.6 35.3
derivatives at the B3LYP/6-31G(d) level. These frequencies 6-31G(d)
were used to verify genuine minimum, transition states, and CASPT2/6-31G(d)//B3LYP/ 35.8
were used to compute zero-point vibrational energy corrections. 6-31G(d)
These frequencies were also used to compute enthalpy and free a
CASPT2/6-31G(d)//B3LYP/6-31G(d) predictions are adopted from
energy corrections. For small polyenes (P1-P4), we applied the previous theoretical studies (refs 32 and 34). b B3LYP/6-31G(d) pre-
CASMP2/6-311G(d)//CASSCF/6-31G(d) theory. The B3LYP dictions adopted from Bernardi et al. (ref 32).
results of these polyenes were then compared with those of
CASMP2 and CASPT2. CASMP2 and CASPT2 theories. It is observed that ∆Eq
The singlet transition states of polyenes or carotenes have a (difference of electronic energy plus zero-point vibrational
strong diradical, open-shell singlet character and hence should energy) and ∆Hq (difference of enthalpy at 298 K) predicted
be defined at least by a two-determinant wave function. Within using B3LYP are very close to those predicted using the
unrestricted DFT, this issue is resolved by spin correction, which CASPT2 theory. In contrast, the CASMP2 predictions overes-
was suggested by Yamaguchi et al.33 The corrections turn out timate the barriers by several kilocalories per mole. In all the
to be very small (within 1 kcal/mol) in all cases. cases, the differences between the values obtained using B3LYP
and CASPT2 are less than 1 kcal/mol. Our B3LYP/6-31G(d)
3. Results and Discussion predicted ∆Eq and ∆Hq to be slightly smaller than those
3.1. Polyenes. In the case of small polyenes, the energy predicted by Bernardi et al.;32 this difference is attributed to
barriers computed using B3LYP/6-31G(d) for rotating the central the inclusion of spin correction in the singlet transition states.
double bonds are compared with those computed using high- In addition, vertical (singlet and triplet energies computed at
level ab initio theories (see Table 1). It should be noted that the S0 geometry) and adiabatic (energies computed at the S0
the rotational barriers predicted for P1-P4 using DFT are in and T1 optimized geometries, respectively) singlet-triplet
reasonably good agreement with those predicted using the energy separations (∆E′S-T and ∆ES-T) predicted using B3LYP
12160 J. Phys. Chem. B, Vol. 112, No. 38, 2008 Guo et al.

TABLE 2: Barriers of Activation Energy (∆Eq), Enthalpy The transition states involved in the all-trans to cis rotations
(∆Hq), and Gibbs Free Energy (∆Gq), the Reaction Energy at both S0 and T1 surfaces of P9, P11, and P13 are illustrated in
(∆E), Enthalpy (∆H), and Gibbs Free Energy (∆G), and the Figure 2 (results of all polyenes are illustrated in the Supporting
Vertical and Adiabatic Singlet-Triplet Energy Separations Information). As can be observed in Figure 2, the geometry of
(∆E′S-T and ∆ES-T) of All-Trans Polyenes (in kcal/mol)
the transition states at the S0 and T1 surfaces are very similar;
Predicted using B3LYP/6-31G(d)a
the most noticeable difference is observed in the case of the
∆Eq ∆Hq ∆Gq ∆E ∆H ∆G ∆E′S-T ∆ES-T torsional angle at the central double bond. The geometries of
S0 the transition states reveal that they exhibit strong diradical
P1 57.4 57.7 (58.1)b 58.0 (65)c 0.0 0.0 0.0 104.1 60.4 nature; therefore, the electronic nature of the transition states
P2 48.7 48.8 48.6 0.0 0.0 0.0 75.4 54.5 at the S0 and T1 states mainly differ by the spin couplings of
P3 40.5 40.4 (38.9)b 41.0 (42.2)d 2.0 2.0 1.9 58.6 41.3 the diradical electrons. With regards to the energy, we found
P4 36.4 36.2 36.5 2.1 2.0 2.0 48.6 33.0 that for polyenes with more than four acetylene units, the
P5 32.4 32.2 (32.1)b 32.5 2.1 2.0 1.8 42.0 27.2
P6 30.0 29.9 30.1 2.2 2.1 2.1 37.3 23.1
transition states of T1 are higher in energy by no more than 1
P7 27.8 27.6 (27.5)b 27.9 2.1 2.1 1.7 33.9 19.9 kcal/mol than those of S0.
P8 26.2 26.0 26.3 2.2 2.2 2.1 31.2 17.4 The predicted singlet-triplet energy separations in Table 2
P9 24.7 24.5 (24.5)b 24.8 2.1 2.2 1.4 29.1 15.4 reveal the electronic nature of the polyenes. As a result of the
P10 23.6 23.4 23.7 2.2 2.2 1.9 27.5 13.7 extended π-conjugation system, the relative energy of the T1
P11 22.4 22.2 (22.4)b 22.6 2.1 2.2 0.8 26.1 12.3 state decreases when n increases. We observed that both ∆E′S-T
P12 21.6 21.4 21.7 2.2 2.2 1.4 25.0 11.0 (singlet and triplet energies computed at the S0 geometry) and
P13 20.7 20.5 (21.1)b 20.9 2.1 1.6 2.9 24.1 10.0
P14 20.0 19.8 20.2 2.2 1.6 3.1 23.3 9.0 ∆ES-T (singlet and triplet energies computed at the S0 and T1
P15 19.3 19.1 (20.1)b 19.6 2.2 2.2 0.8 22.7 8.2 geometries) decrease with an increase in n. On the basis of the
studies on polyenes using the Hubbard and Pariser-Parr-Pople
T1
P1 17.8 17.6 17.5 0.0 0.0 0.0
models, Tavan and Schulten demonstrated fairly linear relations
P2 0.0 -0.5 0.6 0.0 0.0 0.0 between the singlet excitation energies and 1/(2n + 1).23
P3 1.0 0.6 1.9 0.3 0.3 0.1 Interestingly, similar correlations were observed for our pre-
P4 4.4 4.0 4.8 2.5 2.1 2.9 dicted values of ∆E′S-T and ∆ES-T (Figure 3).
P5 5.9 5.5 6.3 2.9 2.9 3.0 For the S0 state, the rotational barriers are smaller in extended
P6 7.6 7.2 7.9 2.9 3.0 2.3
polyenes. The above-mentioned trend of ∆Eq versus n in Pn is
P7 8.4 8.0 8.7 3.0 3.1 1.7
P8 9.2 8.9 9.5 3.0 3.1 2.5 attributed to the stabilization of the open-shell singlet diradical
P9 9.7 9.4 10.0 3.0 3.1 2.1 by the polyacetylene units: the larger the unit, the smaller the
P10 10.2 9.9 10.4 3.0 3.1 2.3 barrier. The imaginary vibrational frequencies also decrease with
P11 10.5 10.2 10.7 3.0 3.2 2.0 n (see the Supporting Information). The rotational barriers of
P12 10.8 10.5 11.0 3.0 2.5 3.7 Pn predicted in this work are in good agreement with the
P13 11.0 10.7 11.2 3.0 3.2 1.7 experimental results of Doering et al., which were derived from
P14 11.2 10.9 11.4 3.0 2.6 3.7
P15 11.3 11.1 11.5 3.0 3.2 1.6
the kinetic data of semirigid polyenes.24
a
The rotational barriers for the S0 and T1 states of polyenes
Available experimental results and predictions from previous are plotted against n in Figure 4a. At the S0 state, ∆Eq decreases
predictions are included in parentheses. b Experimental measurement
for the semirigid counterparts of Pn from Doering and Sarma (ref
with an increase in n. This trend is attributed to the stabilization
24). c Experimental rotational barriers (∆Gq) (ref 44). d Experimental of the transition state in larger polyenes. The geometries of the
rotational barriers ∆Gq) (ref 45). transition states of polyenes contain CdC double bonds rotated
to ∼90° (refer to the Supporting Information); therefore, the
more extended π-conjugation systems are capable of stabilizing
are in good agreement with those obtained from the CASPT2 the transition states.
theory but are smaller than those obtained using the CASMP2 In contrast to ∆Eq of polyenes in the S0 state, that of polyenes
theory. at the T1 state increases with n. This seemingly contradictory
The comparison between B3LYP and ab initio theories result is a consequence of the fact that, at larger n, the extended
encourages the application of B3LYP to the computation of the π-conjugation system stabilizes the T1 minima to a larger extent
potential energy surfaces for the isomerizations at the ground than it stabilizes the transition states. This is demonstrated by
state (S0) and first triplet state (T1) of polyenes. All-trans the dependency of ∆ES-T on n, as shown in Figure 4a. Since
polyenes (H-(CHdCH)n-H, Pn) up to P15 are examined using the energies of the transition states for S0 and T1 are nearly
the B3LYP/6-31G(d) method, and the results are summarized degenerate because they are a singlet and triplet pair of open-
in Table 2. shell diradicals, respectively, the rotational barrier in the T1 state
The geometries of singlet and triplet Pn are all planar, the is proportional to n. This correlation explains how the potential
only exception being triplet ethene, which adopts a D2d energy surfaces of the Pn change with the length of polyenes.
geometry. For simplicity, we illustrated only the geometrical With respect to the S0 minima, as n of Pn increases, the relative
parameters of P9, P11, P13, and their central-cis isomers in Figure energies of T1 decrease more drastically than those of the
1. The geometrical parameters of all the polyenes are illustrated transition states (refer to Figure s4 of the Supporting Informa-
in the Supporting Information. Among the singlet polyenes, we tion). Linear correlations are observed when we plot ∆Eq of S0
observed the bond-length alternations and found that the extent and T1 against ∆ES-T. As illustrated in Figure 4b, the rotational
of alternation becomes less apparent for larger polyenes. In barriers of singlet Pn are directly proportional to ∆ES-T, whereas
contrast, the bond lengths of triplet all-trans Pn reveal switching those of the triplet Pn are inversely proportional to ∆ES-T.
between the double and single bonds, particularly at the central An exception to the aforementioned dependency is triplet P1,
region. The longest bond in the T1 state is the central double for which the rotational barrier is the largest among the triplet
bond. polyenes studied. Unlike the other triplet Pn compounds, the
Cis-Trans Isomerization of β-Carotene and Lycopene J. Phys. Chem. B, Vol. 112, No. 38, 2008 12161

Figure 1. Geometrical parameters (in angstroms) of singlet (S0) and triplet (T1) polyenes (P9, P11, P13), and their corresponding central-cis isomers
optimized at B3LYP/6-31G(d) level. Geometrical parameters of S0 and T1 states are shown in the upper and lower entries, respectively.

minimum of P1 adopts a nonplanar, D2d symmetry. Its transition
state, however, has a planar geometry. The electronic structure
of P1 is, therefore, not comparable to that of the other polyenes.
The cis isomers at the S0 and T1 states are higher in energy
than their all-trans counterparts.
3.2. β-Carotene and Lycopene. The potential energy sur-
faces of the isomerizations of all-trans-β-carotene and lycopene
were investigated using the B3LYP approach. For simplicity,
the structures of all the isomers and transition states are
summarized in the Supporting Information. We include the
optimized structures of the two carotenes and their 13-cis and
15-cis isomers in Figure 5. The structures and magnitudes of
the imaginary frequencies for the transition states corresponding
to the all-trans to 13-cis and 15-cis isomerizations are shown
in Figure 6.
The optimized structures of β-carotene and lycopene reveal
the following trends similar to those observed in polyenes; bond-
length alternation at the S0 state and the bond-length inversion
at the T1 state. Bond-length alternation is apparent at the central
part of the compound and is less apparent at the peripheral
region of the conjugated chains. In contrast to the marked
difference between the geometries of S0 and T1, their transition
12162 J. Phys. Chem. B, Vol. 112, No. 38, 2008 Guo et al.

Figure 2. Geometrical parameters (in angstroms) and magnitude of imaginary vibrational frequencies (νimag, in i cm-1) of the isomerization transition
states (TSs) of singlet (S0) and triplet (T1) polyenes (P9, P11, P13) located at B3LYP/6-31G(d) level. The results of S0 and T1 states are shown in the
upper and lower entries, respectively.

Figure 3. Singlet-triplet energy separation (∆ES-T, b, R2 ) 0.9940),

and vertical singlet-triplet energy separation (∆E′S-T, 9, R2 ) 0.9993,),
vs 1/(2n + 1) for polyenes P2-P15. The energies are in kcal/mol.

states (Figure 6) are very similar. For each isomerization, the

transition states at S0 and T1 surfaces are nearly degenerate (vide
infra). The imaginary vibrational frequencies of the S0 state are
higher than their counterparts at the T1 state.
The rotational barriers and energies of isomerization reactions
involved in the isomerizations of all-trans-β-carotene and
lycopene are summarized in Tables 3 and 4. Relative energies
(including zero-point vibrational energy corrections) of all-trans-
β-carotene and lycopene, their mono-cis isomers, and the
transition states of isomerizations at the S0 and T1 states are
schematically illustrated in Figures 7 and 8.
For both β-carotene and lycopene, at their S0 and T1 states,
the lowest rotational barrier occurs at the 13-14 CdC bond,
rather than the 15-15′ CdC bond. This observation is
contradictory, for the experiments performed by Doering’s group
Figure 4. (a) Rotational barrier (∆Eq) of S0 ([) and T1 (2) and the
singlet-triplet energy separation (∆ES-T, 9) vs n; (b) ∆Eq of S0 (0,
R2 ) 0.9971) and T1 (O, R2 ) 0.9793) states vs ∆ES-T, for P2-P15.
The energies are in kcal/mol.
revealed that the rotational rates for the conversion of the all-
trans isomer to the 13-cis isomer of β-carotene are higher than
those for the conversion of its all-trans isomer to the 15-cis
isomer.21 We attribute this result to the stabilization of the
transition state by methyl substitution at the 13th carbon. The
stabilization effect of methyl group on the rotational barrier of
Cis-Trans Isomerization of β-Carotene and Lycopene J. Phys. Chem. B, Vol. 112, No. 38, 2008 12163

Figure 5. Geometrical parameters (in angstroms) of all-trans, 13-cis, and 15-cis isomers of singlet (S0) and triplet (T1) carotenes optimized at
B3LYP/6-31G(d) level. Geometrical parameters of S0 and T1 states are shown in the upper and lower entries, respectively.

an alkene has been studied by Jarowski et al.34 To verify this
proposal, we examined the rotational barriers of trans-2-butene
and 2-methyl-trans-butene at the B3LYP/6-31G(d) level. Indeed,
the energy of the rotational barrier of 2-methyl-trans-butene is
lower than that of trans-2-butene by 1.3 kcal/mol.
At S0, the rotational barrier for the conversion of the all-
trans to mono-cis isomers for both β-carotene and lycopene are
in the following order: 13-cis < 15-cis < 11-cis < 9-cis < 7-cis
(< 5-cis, lycopene). Nevertheless, we see that for all isomers
except for 7-cis rotations, the energies of the rotational barriers
12164 J. Phys. Chem. B, Vol. 112, No. 38, 2008 Guo et al.

Figure 6. Geometrical parameters (in angstroms) and magnitude of imaginary vibrational frequencies (νimag, in i cm-1) of the isomerization transition
states (TSs) of singlet (S0) and triplet (T1) carotenes located at B3LYP/6-31G(d) level. The results of S0 and T1 states are shown in the upper and
lower entries, respectively.
Cis-Trans Isomerization of β-Carotene and Lycopene J. Phys. Chem. B, Vol. 112, No. 38, 2008 12165

TABLE 3: Barriers of Activation Energy (∆Eq), Enthalpy

(∆Hq), and Gibbs Free Energy (∆Gq), the Reaction Energy
(∆E), Enthalpy (∆H), and Gibbs Free Energy (∆G) of
all-trans-β-Carotene into its Cis Isomers, and the Adiabatic
Singlet-Triplet Energy Separations (∆ES-T) of Isomers (in
kcal/mol) Predicted using B3LYP/6-31G(d)
∆Eq ∆Hq ∆Gq ∆E ∆H ∆G ∆Erq a ∆ES-T
S0 13.8b
13-cis 22.6 (27.7)c 22.5 22.7 1.1 (1.3)c 1.0 1.6 21.5 15.3
15-cis 23.2 (28.9)c 23.2 23.1 2.6 (1.7)c 2.6 2.5 20.6 14.4
11-cis 24.2 24.2 23.6 5.3 5.1 5.9 18.9 13.8
9-cis 26.9 26.9 26.8 0.9 0.8 1.2 26.0 14.2
7-cis 27.4 27.3 27.3 5.2 5.1 5.6 22.2 13.3
T1
13-cis 9.1 9.5 8.6 2.6 3.3 1.6 6.5
15-cis 9.7 10.0 9.0 3.2 4.0 1.7 6.5
11-cis 10.9 11.1 11.2 5.3 5.9 4.2 5.6
9-cis 13.4 13.7 12.8 1.3 1.9 0.3 12.1
7-cis 14.0 14.2 13.5 4.6 5.3 3.0 9.4
a
Rotational barrier for the mono-cis to all-trans reverse reaction.
b
∆ES-T of all-trans-β-carotene. c Arrhenius parameters obtained by
Doering et al. (ref 21).

TABLE 4: Barriers of Activation Energy (∆Eq), Enthalpy

(∆Hq), and Gibbs Free Energy (∆Gq), the Reaction Energy
(∆E), Enthalpy (∆H), and Gibbs Free Energy (∆G) of
all-trans-Lycopene into its Cis Isomers, and the Adiabatic
Singlet-Triplet Energy Separations (∆ES-T) of Isomers (in
kcal/mol) Predicted using B3LYP/6-31G(d)
∆Eq ∆Hq ∆Gq ∆E ∆H ∆G ∆Erq a ∆ES-T Figure 7. Potential energy diagrams of all-trans-β-carotene, their
mono-cis isomers, and the transition states of isomerization at the S0
b
S0 11.9 (a) and T1 (b) states. The energies (in kcal/mol) are presented with
13-cis 21.1 20.9 20.5 1.2 1.1 1.4 19.9 13.4 reference to the all-trans isomer of S0.
15-cis 21.9 21.6 21.1 2.8 2.7 2.4 19.1 12.6
11-cis 22.2 21.8 22.2 5.4 5.2 5.8 16.8 11.8
9-cis 24.1 23.9 23.9 1.0 0.9 0.9 23.1 12.5
β-carotene and lycopene are 23.2 and 21.6 kcal/mol, respectively.
7-cis 27.6 27.4 27.7 5.5 5.2 6.1 22.1 11.6 The energy of the central (15-cis) rotational barrier of β-carotene
5-cis 35.4 35.1 35.5 0.2 0.0 1.1 35.2 11.7 is higher than that of P11, whereas the energy of the rotational
T1 barrier of lycopene is lower than that of P11. Despite the fact that
13-cis 9.3 9.1 9.2 2.6 2.8 1.6 6.7 our computed order of reactivities and relative stabilities for all-
15-cis 10.0 9.7 9.7 3.4 3.5 2.4 6.6 trans, 13-cis, and 15-cis isomers of β-carotene are in the same order
11-cis 10.4 10.0 10.9 5.2 5.1 5.3 5.2 with those predicted by Doering et al.,21 our values are ∼5 kcal/
9-cis 12.4 12.1 12.7 1.6 1.5 1.8 10.8
7-cis 15.9 15.5 16.3 5.2 5.0 5.4 10.7 mol higher in energy than those predicted by Doering (Table 3).
5-cis 23.7 23.4 24.5 0.0 -0.1 0.4 23.7 At present, we cannot offer an explanation to justify this noticeable
deviation.
a
Rotational barrier for the mono-cis to all-trans reverse reaction. For both the carotenes, the all-trans isomers are the most stable
b
∆ES-T of all-trans-lycopene.
at the S0 and T1 states. The order of the relative energies of the
mono-cis isomers is also similar for both the electronic states as
of lycopene are higher than those of their counterparts in follows: (5-cis, lycopene) < 9-cis < 13-cis < 15-cis < 7-, 11-cis.
β-carotene. Therefore, at room temperature, the isomerization Thus, thermodynamically, it is easier to rotate a methylated double
rates of lycopene are higher by 1-2 orders of magnitude than bond (5-cis, 9-cis, and 13-cis) than an unmethylated double bond,
those of β-carotene. i.e., 15-cis, 7-cis, and 11-cis. Among the methylated cis isomers,
Despite the fact that both these carotenes possess 11 it is easier to rotate the peripheral ones (5-cis and 9-cis). In contrast,
conjugated π-bonds, the presence of the β-ionone ring in in the case of unmethylated double bonds, the central 15-cis isomer
β-carotene reduces electron delocalization. ∆ES-T of all-trans- is more susceptible to rotation. It has to be noted that, although
β-carotene is 13.8 kcal/mol; in contrast, ∆ES-T of all-trans- 5-cis-lycopene is a thermodynamically stable product, it has an
lycopene is lower by ∼2 kcal/mol. Similarly, ∆ES-T values of extraordinarily high rotational barrier.
the lycopene isomers are lower than those of their β-carotene The relative stabilities of the isomers, as shown in Figures 7
counterparts (see Tables 3 and 4). The low-lying triplet of and 8 and in Tables 3 and 4, are generally in good agreement with
lycopene could be responsible for its significant antioxidative the experimental values. In chloroplasts and solvents, the 9-cis and
effects among carotenes. 13-cis isomers of β-carotene and lycopene were found to be the
On the basis of experimental measurements, Doering and Sarma predominant cis isomers.15 In human serum and tissues, the 9-cis,
extrapolated their results and predicted a rotational barrier (∆Hq) 13-cis, and 15-cis isomers of β-carotene and lycopene are present.13
of 22.4 kcal/mol for semirigid P11.24 Our DFT computation for From the experiments on both pure β-carotene and carrot juices,
the rotational barrier of P11 (22.2 kcal/mol, see Table 2) is in close 13-cis isomer was found to be the dominant cis isomer, whereas
agreement with Doering and Sarma’s prediction. Considering under lighted storage, the accumulation of 9-cis isomer was
β-carotene and lycopene that involve a branched P11 conjugated favored.35,36 In green vegetables and in fruits, 9-cis-β-carotene was
system, the predicted energies of the 15-cis rotational barriers in found to be the major cis isomer.37 In Figure 7, we can observe
12166 J. Phys. Chem. B, Vol. 112, No. 38, 2008 Guo et al.

that the relative energies of the 9-cis and 13-cis isomers are
comparable, whereas the formation of 9-cis involves a larger
rotational barrier. Aman et al. revealed that in heated chloroplast
isolates, pure 13-cis-β-carotene and 13-cis-lutein are the predomi-
nant isomers, whereas 9-cis is the predominant cis isomer in heated
chloroplast.15 This observation is justified as the formation of 13-
cis-β-carotene is kinetically favored, whereas the formation of 9-cis-
β-carotene is thermodynamically favored.
The 11-cis isomer of β-carotene has received less attention as
its spectrum is very similar to that of the all-trans isomer, making
its identification more difficult than other isomers.38 In addition, it
has been pointed out by Hu et al. that the 11-cis isomer would
experience significant steric repulsions, thereby rendering it be
unstable to coexist with other isomers at room temperature.38 Our
results are in accordance with these predictions, i.e., the 11-cis
isomer is the least stable species on both the S0 and T1 potential
energy surfaces. Kinetically, 11-cis-β-carotene has the smallest
barrier toward all-trans-β-carotene and thus can most easily elude
experimental identification. In addition, 7-cis is also an unstable
species. At the ground state, 7-cis-β-carotene is only 0.1 kcal/mol
lower in energy than 11-cis-β-carotene. The rotational barrier of
7-cis-β-carotene, however, is larger than that of 11-cis-β-carotene.
7-cis-β-Carotene was observed by Tsukida and Saiki, whereas 11-
cis-β-carotene was not.39
The thermal isomerization reaction of mono-cis isomers with
all-trans-β-carotene reveals that the experimental isomerization
rate is in the following order: 7-cis < 9-cis < 13-cis < 15-cis
< 11-cis.20,38 The rate of isomerization according to our
prediction is in the order of 9-cis < 7-cis < 13-cis < 15-cis < Figure 8. Potential energy diagrams of all-trans-lycopene, their mono-
cis isomers, and the transition states of isomerization at the S0 (a) and
11-cis. The energy barrier of thermal isomerization of 9-cis-β- T1 (b) states. The energies (in kcal/mol) are presented with reference
carotene to all-trans-β-carotene was 26.0 kcal/mol, which is to the all-trans isomer of S0.
significantly higher than those of other cis isomers. The di-cis
isomers of 9-cis-β-carotene were identified by Kuki et al.20
the experimental results, with the exception for the 9-cis isomer.
Overall, our results are in reasonable agreement with the
According to our prediction, the 9-cis isomer is most stable with
experimental results, in which the composition of isomers in
respect to isomerization. In addition, the relative compositions
natural or heated β-carotene was investigated.
of the isomers after 1000 or 3000 pulses of irradiation for triplet-
The predicted rotational barriers at S0 suggest that, within sensitized photoisomerization are in the order all-trans . 9-cis
hours, the all-trans isomer is in equilibrium with the first three > 13-cis > 15-cis.40
or four lowest energy isomers. Experimentally, when the thermal We can summarize the rule of thumb for evaluating the
isomerization reaction is carried out for 30 min with the all- relative stability of carotene isomers on the basis of our
trans isomer as the starting material, the amounts of 7-cis and computational data: (1) the all-trans isomers are the lowest
11-cis isomers are negligible.20 Although 9-cis-β-carotene has energy isomers; (2) the methylated cis isomers are lower in
the next higher population after all-trans-β-carotene, the very energy than the unmethylated cis isomers, and among them the
large rotational barrier forbids its early observation. The relative peripheral cis isomers are lower in energy; (3) among the
populations of the isomers obtained from our thermodynamic unmethylated cis isomers, central-cis (15-cis) isomers are lower
data are consistent with the experimental observations, wherein in energy. These general features are valid for both S0 and T1
13-cis, 9-cis, and 15-cis isomers are the products formed by potential energy surfaces. Although the information for the
the isomerization of all-trans-β-carotene at 80 °C.20 isomerizations of pure carotenes may not be applied to chlo-
As revealed by Koyama’s group, the 13-cis and 15-cis isomers roplast-bound carotenes, the isomerization processes of the two
of β-carotene are efficiently isomerized toward the all-trans important carotene species examined in this study should provide
configuration upon triplet excitation.1,20 Our predictions (Table 3) valuable insight for future studies.
show that at the T1 state, the 11-cis, 13-cis, and 15-cis isomers are The relative stability of the 9-cis isomers of β-carotene and
most susceptible to isomerization. The theoretical predictions are lycopene may have important physiological implications. As
in good agreement with the observations (11-cis was not included mentioned in the Introduction, cis isomers including 9-cis, 9,13-
in the triplet sensitization experiment). However, our prediction di-cis, and 9,9′-di-cis isomers are the predominant cis isomers
for the isomerization efficiency of the triplet 9-cis isomer remains in blue-green algae and spinach.17 9-cis-β-Carotene was found
questionable when compared with the experimental values. to be the predominant component in Cyt b6f of spinach,18
The triplet-sensitized photoisomerization of the all-trans and whereas 9-cis-R-carotene is bound to the Cyt b6f complex of
mono-cis isomers of β-carotene shows that the decrease of the green algae.19 Zhang et al. have reported the existence of
isomers at the initial stage of irradiation is in the following order: stoichiometrically bound (9-cis or 15-cis) β-carotene in the Cyt
all-trans < 7-cis < 9-cis < 13-cis < 15-cis.40 According to the b6f extracts obtained from a thermophilic bacterium, spinach,
computed rotational barriers, the tendency for isomerization is and a green alga.41 It was suggested that the function of the
9-cis < all-trans < 7-cis < 13-cis < 15-cis < 11-cis at T1. protein-bound β-carotene is to provide protection of the Cyt
Again, we found that the predicted activities agree well with b6f complex against the toxicity of photosynthetically produced
Cis-Trans Isomerization of β-Carotene and Lycopene
O2.41 In vitro experiments of Levin and Mokady suggest that
9-cis-β-carotene has a higher antioxidant potency than that of
the all-trans isomer.42 Liu and Osawa showed that the cis
isomers, especially 9-cis-astaxanthin, exhibit higher antioxidant
activities in vitro than the all-trans isomer.43 Being more stable
and more thermodynamically favored than other cis isomers,
the presence of these 9-cis isomers, particularly in Cyt b6f
complexes, may play important roles in antioxidant functions.
4. Conclusion
We have studied the mono-cis rotations of all-trans polyenes,
β-carotene, and lycopene. The B3LYP method has been demon-
strated to be able to provide reliable predictions for the rotational
barriers of the species in this study. In the case of polyenes, we
found that ∆Eq for Pn isomerization at S0 decreases with an increase
in n; in contrast, ∆Eq increases with n at the T1 state. The difference
is attributed to the fact that, with respect to S0, the relative energy
of T1 decreases at a faster rate than the energy of the transition
state with an increase in n. A linear correlation was observed
between ∆ES-T and 1/(2n + 1) for Pn, which was in agreement
with the prediction of Tavan and Schulten. In addition, we found
that ∆Eq of Pn is linearly proportional to ∆ES-T at S0 and inversely
proportional to ∆ES-T at T1. Therefore, the singlet-triplet energy
separation is a very good indicator for the stability of a polyene
cis-trans isomerization. For carotenes at the S0 state, lycopene
has lower rotational barriers and higher rotational rates than its
β-carotene counterparts. Lycopene has a more low-lying triplet state
than β-carotene, which may be responsible for its enhanced singlet-
oxygen quenching ability. On the basis of our computed data, it
can be suggested that, at room temperature, the low-lying isomers
of carotenes are in equilibrium with the all-trans isomer in the
following order: all-trans > 9-cis > 13-cis > 15-cis. The rule of
thumb for evaluating the relative stability of carotene isomers
derived from this study is as follows: (1) all-trans isomers are the
most stable; (2) the methylated cis isomers are lower in energy
than the unmethylated cis ones, and the peripheral methylated cis
isomers are lowest in energy; (3) among the unmethylated cis
isomers, central-cis is the most stable. Among all the cis isomers,
the 9-cis isomers are extraordinarily stable. The presence of 9-cis
isomers in photosynthetic systems may be attributed to their stability
and antioxidative ability.
Acknowledgment. The authors acknowledge that the National
Science Council of Taiwan, Republic of China, supported this work.
We also thank the National Center for High-Performance Comput-
ing in Taiwan for computer time and facilities.
Supporting Information Available: Geometrical parameters
of compounds and transition states optimized at the B3LYP/6-
31G(d) level. This material is available free of charge via the
Internet at http://pubs.acs.org.
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