Changes in he con en o ee amino acids In
che imoya ui s a a ious ipening
empe a u es
Cambios en el con enido de aminoácidos lib es en u os de chi imoyo a
dis in as empe a u as de madu ación
TORRES, M. N.; LÁZARO, J. J.; PERIAGO, J. L.; GIL, A. and FAUS, M. J.
Depa amen o de Bioquímica y Biología Molecula . Facul ad de Fa macia, Uni e sidad de
G anada. 18071 G anada, España.
RESUMEN
Las a iaciones en el con enido de aminoácidos lib es en u os du an e la madu a
ción es un ema de in e és debido a la elación de es os compues os con su olo , sabo
y es uc u a. Es e es udio se ha diseñado pa a de e mina los cambios en aminoácidos
lib es du an e la madu ación de los u os del chi imoyo (Annona che imolia a o Fino de
Je e), e aluando la in luencia de la empe a u a de almacenamien o después de la eco
lección. Ex ac os me anólicos de epica pio y mesoca pio del u o se usa on pa a la
cuan i icación de los aminoácidos lib es, los cuales ue on analizados po c oma og a ia
de in e cambio iónico. Ci ulina y p olina se de ec a on en can idades ela i amen e al as.
Glu ama o, glu amine, aspa a o, aspa agina y los no p o einogené icos au ina, ácido y
aminobu í icoy ácido a-aminoadípico ue on encon ados en concen aciones que a ia
on en e 2-6% del o al del con enido de aminoácidos lib es. La madu ación inc emen ó
el con enido de aminoácidos, sob e odo de p olina, ci ulina y glu ama o, cuando los
chi imoyos se almacena on a 20 oC o 8 oc. Es e inc emen o no ocu ió cuando la
empe a u a de almacenamien o se man u o a 4 oC, sugi iendo que el daño po ío a ec a
ma cadamen e a la ía de sín esis de la p olina a pa i de glu ama o.
Palab as cla e: Aminoácidos. Annonacea. Chi imo�a. Madu ación.
ABSTRACT
The a ia ions in he con en o ee amino acids in ui s du ing ipening is a ma e
o in e es since hese compounds a e ela ed o hei la ou , as e and s uc u e. This
s udy was designed o de e mine he changes o ee amino acids in ipening che imoya
ui s (Annona che imolia a . Fino de Je e), e alua ing he in luence o he s o age
empe a u e a ie ha es ing. Me hanolic ui ex ac s o epica p and mesoca p we e
used o quan i ica ion o ee amino acids which we e analyzed by ion-exchange
ch oma og aphy. Ci ulline and p oline we e de ec ed in ela i ely high amoun s. Glu ama e,
glu amine, aspa a e, aspa agine and he non p o einogenic au ine, y-aminobu y ic and y
aminoadipic acids we e ound in concen a ions ha a ied be ween 2-6% o he o al ee
amino acids con en . Ripening inc eased he con en s o amino acids, mainly p oline,
A s Pha maceu ica, 36:1; 51-58,1995
52 TORRES, M. N., LÁZARO, J. J., PERJAGO, J. L., GIL, A. and FAUS, M. J.
ci ulline and glu ama e when che imoya s o age a 20 oC o 80 oC. These inc eases did
no occu when he s o age empe a u e was main ained a 4 oC sugges ing ha chilling
inju y ma kedly a ec s he syn hesis pa hway o p oline om glu ama e.
Key wo ds: Amino acids. Annonaceae. Che imoya. Ripening.
Recibido: 20-10-94.
Acep ado: 16-11-94.
BIBLID [0004-2927(1995) 36:1; 51-58]
INTRODUCTION
Che imoya ui is one o he iches sou ces o ee amino acids, accoun ing
o 10.4% o ui d y weigh . (1, 2). In he las ew yea s, a g ea deal o
in e es has be en cen e ed on he ee amino acid con en o so ne ui s (3-6).
Ven u a e al. (7) ha e examined he suga apple Annona squamosa, which
p esen s he unusual and in e es ing me abolical cha ac e is ic ha y-aminobu y ic
acid and ci ulline p edomina e among he o al amino acids, inc1uding a ginine
and o ni hine. In he sou sop (Annona mu ica a) p oline and y-aminobu y ic
acid a e he majo amino acids, oge he wi h nine o he s which appea in
smalle amoun s (7). Recen ly Touche e al. ha e epo ed ha Annona c .
c assi lo a and Annona paludosa a e cha ac e ized by a high con en o basic
amino acids, pa icula ly his idine, lysine, a ginine and omi hine. (3).
The a ia ions in he con en o ee amino acids in ela ion o me abolic
changes occu ing du ing g ow h and ipening, p ocesses ha a e o undamen
al impo ance in con olling he pos -ha es beha iou o ui s, ha e ecei ed
li le a en ion. A p esen he e a e no da a conceming he quali a i e and
quan i a i e pa ems o ee amino acids in ipenig che imoya ui s.
The aim o his s udy was o de e mine he changes in he con en o ee
amino acids o ipening che imoya ui s a e ha es a he g een- ipe s age. To
e alua e he in luence o chilling inju y, che imoya ui s we e s o ed a 8 oC
and 4 oC and he changes in he ee amino acid pa ems we e compa ed wi h
hose ob ained in ui s s o ed a 20 oC.
MATERlALS AND METHODS
Plan ma e ial
Uni o m weigh (500 + 50 g) che imoya ui s (Annona che imolia Mill) o
he "Pino de Je e" a ie y we e ha es ed a he g een- ipe s age and subsequen ly
ipened unde one o he ollowing empe a u es: 20 oC, 8 oC o 4 oC, and a
85% ela i e humidi y (RH).
A s Pha maceu ica. 36:1; 51-58. 1995
CHANGES IN THE CONTENT OF FREE AMINO ACIDS IN CHERIMOY A FRUITS ... 53
The ui s we e s o ed o 12 o 40 days depending on he ipening empe a u e.
Sampling o che imoyas o amino acid analysis was done a 4 day in e als
du ing ipening.
Amino acid analysis
The ui s we e washed wi h ap wa e and d ied wi h il e pape . Mesoca p
was sepa a ed o he epica p by a s ainless s eel spa ula and seeds we e emo
ed. Each issue was cu in o small pieces and weighed. Samples homogeniza ion
we e ca ied ou sepa a ely wi h app oxima ely 100 g o mesoca p and 50 g o
epica p. Two olumes o 80% me hanol in wa e con aining 0.2% sodium
disulphi e by epica p o mesoca p weigh we e added. Suspensions we e ob ained
by disin eg a ion using a So all Omnimixe homogenize o 5 mino The
suspensions we e il e ed wice h ough ou laye s o su gical gauze. The
me hanolic ex ac was cen i uged a 1,200 g and acuum concen a ed a 40-
50 oC.
Final olumes o epica p and mesoca p ex ac s we e espec i ely adjus ed
o 100 mI. and 50 mI Samples o amino acid analysis we e dilu ed i e imes
wi h a bu e pH 2.1, con aining 250 nmoles o no leucine as in e nal s anda d.
Amino acids we e de e mined by ion-exchange ch oma og aphy using a
Ch omaspeck-J-180 au oma ic amino acid analyze as desc ibed p e iously (8).
RESULTS
Table 1, 2 and 3 espec i ely show he con en s o mesoca p and epica p
ee amino acids o che imoya ui s ipened a 20 oC, 8 oC, and 4 oC a e
ha es a he g een- ipe s age.
The majo ee amino acids de ec ed in bo h issues we e ci ulline and
p oline. The concen a ion o ci ulline a.ccoun ed o 11-65% and 12-44% o
o al ee amino acids in epica p and mesoca p espec i ely. The ela i e con en s
o p oline a ied be ween 11-58% and 16-60%.
Acidic amino acids and hei amides as well as se ine we e also de ec ed in
ela i ely high amoun s (2-6%). On he con a y, a oma ic and basic amino
acids, excep o ni hine, we e p esen in low amoun s. O ni hine ep esen ed 2-
20% o o al ee amino acids bo h in epica p and mesoca p o che imoya ui s.
B anched chain and neu al alipha ic amino acids we e always p esen in he
ui s bu hei le els we e e y low. Th eonine and hyd oxyp oline we e also
de ec ed in low amoun s.
The non-p o einogenic amino acids, au ine, y-aminobu y ic and a-aminoadipic
we e egula ly ound in quan i ies ha a ied be ween 2-5% o he o al ee
A s Pha maceu ica, 36:1; 51-58, 1995
54 TORRES, M. N., LÁZARO, J. J., PERJAGO, J. L., GIL, A. and FAUS, M. J.
T ABLE 1.-Mesoca p and epica p ee amino acid con en s o che imoya ui s ipened a 20
°C-22 oC and 85% ela i e humidi y a e ha es ing a he g een- ipe s age .
Epica p Mesoca p
Amino acid Day a e ha es ing Days a e ha es ing
1 4 8 12 1 4 8 12
TAU 0. 42 1. 38 0. 39 1. 33 0. 51 0.11 0.45 0. 52
a-AA 0. 42 0.52 0.70 1. 03 0.17 0. 21 0. 60 0. 45
y-AB 0.33 0. 19 0.68 0. 52
ASP 0. 59 0. 31 0. 14 0. 30 0. 05 0. 15 0. 18 0. 12
THR 0.21 0. 19 0. 31 0.48 0. 12 0. 08 0. 17 0. 14
SER 0. 47 0. 26 0. 22 0. 46 0. 19 0.10 0. 16 0. 16
ASN 0. 82 0. 41 0. 65 . 0. 88
GLU 0. 11 0. 28 0. 32 0. 50 0. 16 0. 04 0.07 0. 06
GLN 0.38 0.71 0. 95 1. 43 0.06 0. 07 0. 07 0. 04
PRO 4.90 12.60 2.41 13.42 1.91 4.90 17. 12 7. 20
OH-PRO 0. 17 0. 21 0. 36 0. 56 0. 09 0. 15 0. 16 0.18
GLY 0. 11 0. 08 0. 16 0. 10 0. 06 0. 08 0. 18 0. 15
ALA 0. 10 0. 19 0. 21 0. 17 0. 21 0. 17 0. 14 0. 11
CIT 11. 82 14. 33 15. 83 14. 21 3. 81 1. 72 7. 30 6. 00
VAL 0. 25 0. 22 0. 58 0. 58 0. 11 0. 06 0. 23 0. 20
ILEU 0. 45 0. 23
LEU 0. 25 0. 20 0. 04 0. 04 0. 06 0. 11
TYR 0. 06 0. 15 0.11 0. 25 0. 07 0.08 0. 11 0. 15
PHE 0.27 0. 10 0. 32 0. 62 0.17 0.17 0. 14 0. 10
TRP 0.08 0. 72 0. 14 0. 23 0. 03 0. 08 0. 13 0. 05
HIS 0. 06 0. 08 0. 10 0.06
ORN 1. 21 1. 84 1. 60 2. 21 0. 90 0. 45 1. 05 1.30
LYS 0. 21 0. 28 0. 32 0. 36 0. 15 0. 07 0. 15 0. 18
ARG 0. 00 0.01 0.02 0. 03
-= No de ec ed.
* Resul s a e exp essed as mean o ou de e mina ions in �moles o ee amino acids pe g
o esh issue.
amino acids. Ci ulline was egula ly de ec ed in highe concen a ions in epica p
han in mesoca p wi h independence o he ipening empe a u e. P oline was
also ound ele a ed in epica p wi h he inc eases mo e e iden a 8 oC and 4 oc.
A 20 oC he concen a ion o glu ame e was inc eased in he epica p compa ed
o ha o he mesoca p bu i did no p esen signi ican di e ences be ween
bo h issues a 8 oC and 4 oc.
The concen a ions o ci ulline and p oline we e inc eased a 8 oC and
dec eased a 4 oC in ela ion o hose o 20 oC in bo h epica p and mesoca p,
and glu amine and se ine we e ound a highe le els a 8 oC and 4 oC han a
20 oC.
A s Pha maceu ica, 36:1; 51-58, 1995
CHANGES IN THE CONTENT OF FREE AMINO ACIDS IN CHERIMOYA FRUITS ... 55
TABLE 2.-Mesoca p and epica p ee amino acid con en s o che imoya ui s ipened a 8 oC
and 85% e1a i e humidi y a e ha es ing a he g een- ipe s age .
Epica p Mesoca p
Amino acid Day a e ha es ing Days a e ha es ing
1 8 16 20 1 8 16 20
TAU 0.43 0.66 0.53 0.79 0.57 0.41 0.40 0.42
a-AA 1.21 1.37 0.82 3.08 1.21 0.85 0.92 2.05
y-AB 0.65 0.33 0.74 0.58
ASP 0.46 1.05 0.56 0.81 0.47 0.32 0.44 0.37
THR 0.22 0.58 0.31 0.75 0.37 0.33 0.54 0.60
SER 0.59 1.05 0.64 1.24 0.58 0.45 0.53 0.67
ASN 1.16 1.46 0.45 0.62 2.05 1.67 0.61 1.05
GLU 3.20 2.90 1.39 2.03 1.87 1.02 0.80 0.82
GLN 2.10 1.91 1.10 2.10 6.42 2.97 1.95 2.25
PRO 23.95 22.71 25.42 24.82 38.05 15.21 19.25 9.75
OH-PRO 0.31 0.24 0.32 0.38 0.23 0.46 0.69 0.32
GLY 0.17 0.22 0.15 0.10 0.02 0.01 0.05 0.07
ALA 0.21 0.34 0.15 0.17 0.32 0.41 0.20 0.18
CIT 20.92 19.21 15.13 24.08 8.72 11.33 7.40 7.52
VAL 0.32 0.47
ILEU 0.26 0.25 0.19 0.25 0.30 0.42 0.71
LEU 0.05 0.04 0.04 0.08 0.07 0.17 0.06 0.07
TYR 0.02 0.05 0.03 0.04
PHE 0.10 0.18 0.15 0.14 0.03 0.17 0.08 0.11
TRP 0.02 0.12 0.03 0.06
HIS 0.19 0.15 0.24 0.14 0.05 0.10 0.12 0.21
ORN 1.35 2.95 1.25 0.82 1.21 2.05 1.08 4.10
LYS 0.60 0.71 0.23 0.35
ARG 0.51 0.50 0.40 0.45 0.16 0.23 0.24 0.31
- = No de ec ed.
* Resu1 s a e exp essed as mean o ou de e mina ions in Jlmo1e s o ee amino acids pe g
o esh issue.
DISCUSSION
The amino acid p o ile o che imoya du ing ipening a e ha es a he
g een- ipe s age was cha ac e ized by a high con en o ci ulline and p oline
and mode a e concen a ions o omi hine, glu ama e, aspa a e, glu amine and
aspa agine and low le els o b anched chain a oma ic and basic amino acids.
So ne opical Annonaceae con ain high le els o basic amino acids in he
lea es as well as in he ui s, ci ulline and omi hine being he mos equen ly
isola ed (3). Fu he mo e, suga apple also con ain high le els o ci ulline (6)
and many o he ui s no ela ed o che imoyas a e also cha ac e ized by he
p esence o high concen a ions o basic amino acids (5, 6, 9, 10). P oline is
A s Pha maceu ica, 36:1; 51-58,1995

56 TORRES, M. N., LÁZARO, J. J., PERlAGO, J. L., GIL, A. and FAUS, M. J.
TABLE 3.-Mesoca p and epica p ee amino acid con en s o che imoya ui s ipened a 4 oC
and 85% ela i e humidi y a e ha es ing a he g een- ipe s age
Epica p Mesoca p
Amino acid Day a e ha es ing Days a e ha es ing
1 12 24
32
1 12 24 32
TAU 0.95 0.48 0.98 0.83 0.26 0.39 0.63 0.03
a-AA 0.75 0.23 1.62 1.21 0.44 0.20 0.65 0.24
y-AB 0.45 0.40 0.82 2.71 0.16 0.44 0.55 0.42
ASP 0.98 0.70 2.15 1.06 0.97 0.81 0.75 0.31
THR 0.17 0.10 0.38 0.44 0.26 0.13 0.29 0.10
SER 0.68 0.35 1.41 l.l0 0.68 0.44 0.78 0.39
ASN 0.27 l.l2 0.61 0.50 0.23 0.13 0.23 0.22
GLU 1.40 0.91 3.05 l.l0 1.87 1.39 1.71 0.78
GLN 2.01 3.20 1.42 1.56 1.88 0.80 1.30 1.07
PRO 1.22 0.32 3.10 2.58 0.88 0.37 0.65 1.17
OH-PRO 0.98 0.65 1.85 2.02 0.26 0.37 1.07 0.10
GLY 0.12 0.17 0.08 0.15 0.26 0.05 0.10 0.11
ALA 0.57 0.25 0.93 1.04 0.24 0.29 0.58 0.28
CIT 11.20 6.62 3.95 19.35 2.48 3.67 2.36 5.72
VAL 0.12 0.17 0.22 0.34 0.10 0.03 0.05 0.05
ILEU 0.25 0.26 0.29 0.03 0.05 0.15 0.13 0.06
LEU 0.05 0.06 0.06 0.06 0.02 0.02 0.03 0.09
TYR 0.13 0.28 0.21 0.20 0.10 0.05 0.15 0.04
PHE 0.61 0.07 0.52 0.38 0.12 0.05 0.06 0.22
TRP 0.27 0.12 0.26 0.14 0.02 0.04 0.02 0.02
BIS 0.01 0.15 0.05 0.06 0.01 0.04 0.07 0.09
ORN 0.17 2.65 0.84 0.94
LYS 0.04 0.05 0.04 0.04
ARG 0.05 0.07 0.25 0.06 0.04 0.00 0.04 0.32
- = No de ec ed.
* Resul s a e exp essed as mean o ou de e mina ion in ¡ moles o ee amino acids pe g o
esh issue.
usually p esen in ui s du ing he ipening p ocess (11) and he SOUTSOP, a kind
o che imoya, con ains his amino acid a he highes le els among he s udied
Annonaceae (7). y-aminobu y ic acid was p esen in che imoya bu i s le el was
only mo e han 2% o he o al amino acids in con as o he esul s epo ed
o o he Annonaceae (6).
Tbe ipening p ocess has been obse ed o be in ima ely associa ed wi h he
me abolism o a ginine, p oline and glu amic acid. The p oline le el in pa icu
la being inc eased a he heigh o he ipening p ocess (11).
In OUT s udy p oline and ci ulline as well as glu ama e inc eased quickly
a e ha es in epica p and mesoca p o che imoya ui s. Tbe inc ease in
A .� Pha mn .p.ll i .n. �6:1: 'il-'iR lQQ'i
CHANGES IN THE CONTENT OF FREE AMINO ACIDS IN CHERIMOY A FRUITS ... 57
p oline was no co ela ed wi h a de c ease in a ginine and glu ama e as i has
been desc ibed in g apes (6). Howe e , Pool el al. (12) ha e eco ded a ise in
he le els o bo h a ginine and p oline du ing long e m s o age.
The epica p o che imoya was iche han mesoca p in he concen a ion o
ci ulline, p oline and glu ama e. This ac may be ela ed o a mo e ac i e
syn hesis pa hway in he i s issue since p oline is one o he p incipal compounds
in he main enance o he plan cell wall s uc u e.
When he ipening p ocess o he che imoyas was ca ied ou a 8 oC, we
ob ained he highes concen a ions o ci ulline, glu ama e and p oline, bo h
o epica p and mesoca p. I seems ha dec easing in empe a u e o ipening is
ela ed o an in c ease in he ac i i y o he enzy nic sys ems o p oduce mo e
p oline. Howe e , chilling inju y occu ing a 4 oC ma ked1y dec eased he
le els o ci ulline, glu ama e and p oline bo h in epica p and mesoca p. F om
he da a ob ained in his s udy we may conclude ha he ipening p ocess and
he beginning o he ui senescence in che imoya a e co ela ed wi h inc eases
in he le els o p oline, ci uline and glu ama e, which appea s o be me abolically
associa ed, and ha chilling inju y, conside ably a ec s he accumula ion o
amino acids ha no mally occu du ing he ipening pe iodo
ACKNOWLEDGEMENTS
This wo k has been suppo ed by a g an om Caja Gene al de Aho os de
G anada, G anada (Spain).
REFERENCES
(1) MANNINO, S. and AMELOTTI, G. (1974). La i is a i aliana delle so anze g asse,
51:111-112.
(2) PERIAGO, J. L., TORRES, M. M., SÁNCHEZ-MEDINA, L., GIL, A. and FAUS, M. J.
(1994). A s Ph., 35:315-326.
•
(3) TOUCHE, A., DESCONCLOIS, 1. F., JACQUEMIN, H., LELIEVRE, Y. and FORJACS,
P. (1981). Plan es medicinales e phy o hé apie, 15:4-9.
(4) DESMAlSON, A. M., LE ROUX, M. T. and TIXIER, M. (1977). Bull Soc Pha m
Bo deaux, 116:65-74.
(5) OUPADISSAKOON, C., YOUNG, C. T. and MOZINGO, R. W. (1980). Peanu Sci.,
7:55-60.
(6) PANDEY, R. M., RAO, M. M. and SINGH, R. M. (1974). Scien ia Ho , 2: 383-388.
(7) VENTURA, M. M. and LIMA, H. F. (1961). Phy on, 17:39-43.
(8) RANK-HILGER (1978). "Anali ical Ins umen s o Indus y and Resea ch". Technical
Li e a u e, Wes wood, Ma ga e, Ken CT94JL, England.
(9) LAFON-LAFOURCADE, S. and GUIMBERTEAU, G. (1962). Vi is, 3:130-135.
(10) KLIEVER, W. M. (1969). J. Food. Sci., 34:274-278.
A s Pha maceu ica, 36:1; 51-58,1995
58 TORRES, M. N., LÁZARO, J. J., PERIAGO, J. L., GIL, A. and FAUS, M. J.
(11) PEYNAUD, E. and RIBEREAU-GAYON, P. (1971). "The g ape". In: A. C. Hu1me (Ed.),
The Biochemis y 01 ui s and hei p oduc s. Vol. II. Academic P ess, London, pp. 171-
205.
(12) POOL, R. M., WEAVER, R. J. and KLIWER, W. M. (1972). J. Ame . Soco Ho . Sci.,
97:67-70.
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