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LWT 39 (2006) 10721079


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Nutritional properties of tempeh our from quality


protein maize (Zea mays L.)
E.O. Cuevas-Rodr gueza, N.M. Verdugo-Montoyaa, P.I. Angulo-Bejaranob,
J. Milan-Carrilloa,e, R. Mora-Escobedoc, L.A. Bello-Perezd,
J.A. Garzon-Tiznadoe,f, C. Reyes-Morenoa,e,
a

Maestra en Ciencia y Tecnologa de Alimentos, Facultad de Ciencias Qumico Biologicas, Universidad Autonoma de Sinaloa (FCQB-UAS),
Lichis #1986, La Campina, 80 060 Culiacan, Sinaloa, Mexico
b
Ingeniera Bioqumica, FCQB-UAS
c
Departamento de Graduados en Alimentos, Escuela Nacional de Ciencias, Biologicas, Instituto Politecnico Nacional
d
Centro de Desarrollo en Productos Bioticos, Instituto Politecnico Nacional, Yautepec, Morelos, Mexico
e
Programa Regional del Noroeste para el Doctorado en Biotecnologa, FCQB-UAS
f
Centro de Investigaciones Regionales del Noroeste, Instituto Nacional de Investigaciones Forestales Agrcolas y Pecuarias, Valle de Culiacan,
Sinaloa, Mexico
Received 4 April 2005; received in revised form 30 June 2005; accepted 30 June 2005

Abstract
The objective of this investigation was to evaluate physico-chemical and nutritional properties of tempeh our from a quality
protein maize (QPM). In comparison to untreated QPM, the QPM tempeh our showed a higher (Pp0:05) gelatinization
temperature (81.7 vs 73.9 1C), and resistant starch (4.24 vs 1.9 g/100 g dry our), and a lower (Pp0:05) gelatinization enthalpy (1.94
vs 2.74 J/g) and total starch content (56.9 vs 62.6 g/100 g dry our). The essential amino acids (EAAs) content of raw QPM our was
improved by the solid-state fermentation process. The contents of His, Ile, and Leu increased (Pp0:05) in 0.81, 0.52, and 1.46 g/
100 g protein, respectively. The total sulphur and total aromatic EAAs increased (Pp0:05) in 0.55 and 3.45 g/100 g protein,
respectively. In untreated QPM our, the rst and second limiting EAAs were Lys and Trp, with EAAs score of 0.72. First and
second limiting EAAs in QPM tempeh our were Trp and Lys, with an EAAs score of 0.84. The SSF process increased (Pp0:05)
nutritional indicators as follows: protein efciency ratio (PER) from 1.78 to 2.10, calculated PER from 1.43 to 1.74, and protein
digestibility corrected amino acid score from 0.55 to 0.83. It is concluded that based mainly on its nutritive value, fermented our
may be considered for the fortication of widely consumed cereal-based food product (tortillas, bread, cookies, atoles).
r 2005 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.
Keywords: Nutritional; Quality protein maize; Tempeh

1. Introduction
Maize (Zea mays L) is the third most important food
crop in the world and a major source of energy, protein,
Corresponding author. Maestr a en Ciencia y Tecnolog a de
Alimentos, Facultad de Ciencias Qu mico Biologicas, Universidad
Autonoma de Sinaloa (FCQB-UAS), Lichis #1986, La Campina, 80
060 Culiacan, Sinaloa, Mexico. Tel.:+52 66772136615;
fax: +52 6677 13 66 15.
E-mail address: creyes@uas.uasnet.mx (C. Reyes-Moreno).

and other nutrients for both human and livestock.


Maize contains 713 g/100 g proteins (d.m.). However,
the quality of maize proteins is poor, because they are
decient in the essential amino acids (EAAs) lysine and
tryptophan (Paredes-Lopez, Serna-Sald var, & GuzmanMaldonado, 2000). Due to the economic importance of
maize, genetic improvements have played a key role in
the development of genotypes that could grow in a wide
range of environment, rainfall, and altitudes (CIMMYT, 1985). In Mexico, extensive eld trials have been

0023-6438/$30.00 r 2005 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.lwt.2005.07.003

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carried out at the International Maize and Wheat


Improvement Center (CIMMYT) to identify the most
productive maize cultivars, high in lysine and tryptophan contents (Ortega & Bates, 1983). Through back
crossing and several recurrent selections, maize breeders
of CIMMYT and the National Research Institute for
Forestry, Agriculture and Livestock (INIFAP) have
successfully developed 26 hybrids and cultivars, similar
in yield and other important agronomic properties to
normal maize. These new high-quality protein genotypes
are collectively called quality protein maize (QPM)
(INIFAP, 1999). Solid-state fermentation (SSF) process
represents a technological alternative for improving the
nutritional value of a great variety of legumes and
cereals maintaining acceptable sensory properties. Tempeh is a traditional fermented food produced by SSF of
soybeans. Usually, the fermentation process is performed by Rhizopus strains belonging to the species
Rhizopus oligosporus, R. oryzae, and R. stolonifer. An
important function of the strains in the fermentation
process of soybeans is the synthesis of enzymes, which
hydrolyse its constituents and contribute to the development of a product with desirable texture, avour, and
aroma. Enzymatic hydrolysis also may decrease or
eliminate antinutritional factors; consequently, the
nutritional quality of the fermented product may be
improved (Hachmeister & Fung, 1993; Paredes-Lopez &
Harry, 1988). The potential of using SSF to improve the
nutritional quality of maize in developing countries has
been evaluated (Egounlety & Aworh, 2003; Mugula,
1992; Mugula & Lyimo, 2000). There is, however ,still a
need for more information on the nutritional effects of
using SSF on QPM maize. The objective of this work
was therefore to evaluate the nutritional properties of
QPM tempeh our made by SSF.

Cardenas-Valenzuela, and Reyes-Moreno (2004), with


minor modications. Whole seeds of QPM were soaked
at 25 1C for 16 h in four volumes of 0.09 mol/l an acetic
acid solution (pH 3.1). The seeds were then drained and
cooked at 90 1C for 30 min, cooled at 25 1C, and packed
in perforated polyethylene bags (15  15 cm). A suspension of R. oligosporus (1  106 spores/l) was used to
inoculate the bags. SSF was carried out applying a
fermentation temperature of 35.4 1C for a fermentation
time of 54.6 h. The resulting QPM tempeh was dried at
52 1C for 12 h, cooled at 25 1C and milled (UD, Cyclone
Sample Mill, UD Corp, Boulder, CO, USA) to pass
through an 80-US mesh (0.180 mm) screen. QPM
tempeh our was kept in tightly sealed containers at
4 1C until used.

2. Materials and methods

2.2. Methods

2.2.4. Particle size index (PSI)


Flour samples of 100 g were placed in a series of US
standard sieves (WS Tyler Inc., Mentor, OH, USA) with
the following sizes: No. 40 420 mm; No. 60 318 mm,
No. 80 180 mm, No. 100 150 mm. Sieves were
shaken by a Ro-Tap machine (WS Tyler Inc., Meter,
OH, USA) for 10 min. The material retained on the
sieves was expressed as percent over. To complete the
particle size index
P of ours, the following formula was
applied: PSI aibi, where ai is the percentage of overs
on sieve i, and bi the coefcient relative to sieve i. The bi
values for sieves numbers 40, 60 and 80 were 0.4, 0.6 and
0.8, respectively. Over from sieve No. 100 and the pan
were added and on overall bi 1.0 was assumed
(Bedolla & Rooney, 1982).

2.2.1. Manufacture of QPM tempeh flour


Tempeh our was made by a procedure described
by Cuevas-Rodr guez, Milan-Carrillo, Mora-Escobedo,

2.2.5. Bulk density (rA)


The ground samples were placed in a known volume
stainless cylinder until topped at 25 1C. The device was

2.1. Materials
The QPM (Z. mays L) V 537 variety was obtained
from the National Research Institute for Forestry,
Agriculture and Livestock (INIFAP) Culiacan, Experimental Station, Sinaloa, Mexico. The grains were
harvested, shelled, cleaned and stored in tightly sealed
containers at 4 1C until used. R. oligosporus was
obtained from the Microbiology Laboratory, National
School of Biological Sciences, National Polytechnical
Institute (Mexico, DF).

2.2.2. Proximate composition


The following AOAC methods (1998) were used to
determine proximate composition: drying at 105 1C for
24 h for moisture (method 925.098); incineration a
550 1C for ash (method 923.03); defatting in a soxhlet
apparatus whit 2:1 chloroform/methanol, for lipids
(method 920.39C with minor modications); and
microKjeldahl for protein (Nx6.25) (method 960.52).
Carbohydrate content was estimated by difference.
2.2.3. Total colour difference (De)
The surface colour of samples was measured using
a Minolta Model CR-210 colour difference meter
(Minolta LTD, Japan). L (0 black, 100 white), a
(+value red, value green) and b (+value
yellow, value blue) were recorded. The L, a, and b
values of a white standard tile used as reference were
97.63, 0.78 and 2.85, respectively. DE was calculated as
DE [(DL)2+(Da)2+Db)2]1/2, where DL LstdLsample,
Da astdasample, Db bstdbsample.

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E.O. Cuevas-Rodrguez et al. / LWT 39 (2006) 10721079

topped ve times and the our density obtained dividing


the sample mass by the cylinder volume (Moreyra &
Peleg, 1981).
2.2.6. Water activity (aw)
This parameter was determined in 5 g our samples,
tempered at 25 1C, using a Hygrometer Aqua Lab
Model CX-2 (Decagon Devices Inc., Pullman, WA,
USA) which was calibrated with a potassium chloride
satured solution (aW 0:841 at 25 1C). After leaving the
sample for 1 h, the headspace equilibrium was attained
and the readings taken.
2.2.7. Differential scanning calorimetry (DSC)
Thermal analysis was performed using DSC (TA
Instruments 2010, New Castle, DE, USA) previously
calibrated with indium. Powder samples (2 mg, d.m.)
were weighed directly into DSC aluminium pans, and
after addition of deionized water (20 ml), pans were
sealed and allowed to equilibrate for 1 h. The heating
rate was 10 1C/min, from 30 to 120 1C. An empty pan
was used as reference for all measurements. The
parameters evaluated were DH (enthalpy of crystal
fusion) and To (onset temperature of gelatinization).
2.2.8. Total starch (TS)
TS was measured as described by Goni, Garc aAlonso, and Saura-Calixto (1997). Fifty milligram
samples were dispersed in 6 ml of 2 mol/l KOH and
energically shaken at room temperature for 30 min.
After addition of 3 ml of 0.4 mol/l sodium acetate buffer
pH 4.75 and 60 ml of amyloglucosidase (Sigma A9913) the samples were incubated for 45 min at 60 1C in
a controlled shaking water bath. Starch was measured as
glucose with Peridochrom Glucose GOD-PAP (Ref
676543, Boehringer). The conversion factor from
glucose to starch was 0.9.
2.2.9. Resistant starch (RS)
RS was measured using the modied method described by Saura-Calixto, Goni, Bravo, and Manas
(1993). This method uses the principle of determining
RS from insoluble dietary bre. The procedure consists
of enzymatic hydrolysis of starch with heat stable alpha
amylase (Sigma No. 4-3306) followed by proteolytic
degradation with a protease (Sigma No. P-5380) and a
nal hydrolysis of starch with amyloglucosidase (Sigma
A-9913) to yield glucose. The insoluble dietary bre was
obtained after several steps of rinsing and centrifugation. The RS was extracted from the insoluble residue
with 2 mol/l aqueous KOH and retreated with amyloglucosidase oxidase/peroxidase (GOD-POD). RS was
calculated as glucose (mg)  0.9 (conversion factor due
to starch hydrolysis).

2.2.10. Water absorption index (WAI) and water


solubility index (WSI)
The WAI and WSI were assessed as described by
Anderson, Conway, Pfeifer, and Grifn (1969). Each
our sample (2.5 g) was suspended in 30 ml of distilled
water in a tared 60 ml centrifuge tube. The slurry was
stirred with a glass rod for 1 min at room temperature
and centrifuged at 3000  g for 10 min. The supernatant
was then poured carefully into a tared evaporating dish.
WAI was calculated from the weight of the remaining
gel and expressed in grams of gel per grams of solid. The
WSI, expressed in gram of solids per gram of original
solids, was calculated from the weight of dry solids
recovered by evaporating the supernatant overnight at
110 1C.
2.2.11. Dispersabality
It was determined according to Mora-Escobedo,
Paredes-Lopez, and Gutierrez-Lopez (1994). A our
sample of 1 g was suspended in a graduated conic tube
with 10 ml of distillated water and agitated at 1000 rpm
for 5 min.
2.2.12. pH
The pH of our samples was recorded using a pH
meter. Each our (10 g) was suspended in 100 ml of
boiling distilled water (AACC, 1995). After cooling at
room temperature, the slurry was shaken (1500 rpm,
25 1C, 20 min) using an orbital shaker (Cole Parmer
Model 21704-10, Cole Parmer International, USA).
2.2.13. In vitro protein digestibility (IVPD)
The method proposed by Hsu, Vavak, Satterlee, and
Miller (1977) was used to determine IVPD. A multienzyme system, consisting of a mixture of porcine
pancreatic trypsin type IX, bovine pancreatic chymotrypsin type II and porcine intestinal peptidase grade III
(Sigma Chemical Co, St Louis, MO, USA), was utilized.
QPM ours and distilled water were used to prepare
50 ml of an aqueous protein suspension (6.25 g of
protein/l) with pH adjusted to 8.0, while stirring in a
37 1C water bath. The multi-enzyme solution was
maintained in an ice bath. Aliquots (5 ml) of the multienzyme solution were added with stirring to the protein
suspension at 37 1C. The rapid pH drop was recorded
automatically over a 10 min period using a pH meter.
IVPD was calculated from the equation IVPD
210.4618.10X, where X pH after 10 min.
2.2.14. Amino acid analysis
Five to 10 mg of each sample were placed in 2 ml
ampoules containing internal standard (norleucine) and
0.4 ml of a HCl solution (527.9 ml concentrated HCl/l).
The ampoules were evacuated, sealed, and placed in an
oven at 110 1C for 24 h. After hydrolysis, a 20 ml aliquot
of the hydrolisate was withdrawn, dried, hydrated,

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redried, and subjected to derivatization. Samples for the


determination of cysteine were rst oxidized with
performic acid at 25 1C for 18 h (Hirs, 1967). Performic
acid was removed with the aid of an evaporative
centrifuge and the samples hydrolysed as described
above. The tryptophan content was determined in a
separate analysis. The samples were hydrolysed in
polypropylene tubes in KOH solution (235.62 g KOH/
l) containing 10 g of thiodiglycol, at 110 1C for 18 h
(Hugli & Moore, 1972). After hydrolysis, the KOH was
neutralized with a perchloric acid solution (249.11 g of
perchloric acid concentrated/l). The supernatant was
removed and adjusted to pH 3 with dilute acetic acid,
and 50 ml aliquot was used for derivatization. Quantitation was achieved using a Pierce Standard H amino acid
calibration mixture that was supplemented with tryptophan. The amino acid analysis was performed using the
Pico-Tag system (Waters, Milford, MA). After hydrolysis, aliquots were dried, mixed with 10 ml of ethanol:
water:triethylamine (2:2:1), dried again and reacted with
20 ml phenylisothiocyanate reagent (ethanol:water:
triethylamine:phenylisothiocyanate, 7:1:1:1) at 25 1C
for 20 min (Cohen & Strydom, 1988). Excess reagent
was removed with the aid of a vacuum pump.
Derivatized samples were dissolved in 0.1 ml of a sodium
acetate solution (11.48 g sodium acetate/l) that had been
adjusted to pH 6.4 with acetic acid. A 10 ml aliquot was
injected onto the column. Tryptophan was analysed on
a Waters C18 reversed-phase column (3.9  150 mm)
(Waters, Milford, MA) using the conditions described
by Buzzigoli et al. (1990). It was necessary to use this
column in order to achieve complete resolution of
tryptophan and ornithine. Ornithine was produced by
alkaline hydrolysis of arginine. Analysis of the other
amino acids was carried out using a Waters C18 column
(3.9  150 mm) with gradient conditions described elsewhere (Bindlingmeyer, Cohen, & Tarvin, 1984).
2.2.15. Protein quality evaluation
Protein quality evaluation of the reference and QPM
ours was performed on 40 growing male Wistar rats,
weighing 4575 g at the beginning of the study. Each
protein diet was tested on eight animals randomly
allocated in individual cages (Eggum, 1973). The cages
were housed in a room at 2071 1C temperature and
55% RH, under 12 h light/12 h dark cycles. Diets were
composed of 10 g protein, 9 g fat, 2 g vitamin mix, 5 g
mineral mix, 5 g cellulose and corn starch to make up
100 g. Corn oil was used as fat source. The vitamin and
mineral mixes were AIN-93-VX and AIN-936-MX, and
were obtained from Harland Tekland Laboratory
Animal Diets (Madison, WI, USA). Sodium caseinate
was used as reference (Eggum, 1973; Linder et al., 1997).
An extra 16 animals were fed on a protein-free diet for
assessment of endogenous nitrogen. Food and deionized
water were given ad libitum. Rats were fed with test

1075

diets containing 10% protein for 3 days preliminary


acclimation period and 28 days for the determination of
protein efciency ratio (PER). Feed intake was recorded
every other day. Weight gain was recorded weekly. Net
protein retention (NPR) was measured for an 8-day
period during test days 1826. Feed and fecal nitrogen
contents were analysed by microKjeldahl (method
960.52) (AOAC, 1998). Apparent digestibility (AD,
%), PER and true digestibility (TD, %) were determined according to Eggum (1973). TD was corrected for
endogenous excretion of nitrogen. The following equations were used:
PER WG=PC;
NPR WG WLPFG=PC;
AD % 100TNintake1826 d
 TNfaeces1826 d =TNintake1826 d ,
TD%
100TNintake1826 d  TNfaeces1826 d
TNfaeces protein free diet1826 d =TNintake1826 d ,

where WG is the weight gain (g), PC the protein


consumed (g), WLPFG the weight loss of protein free
group (g) and TN the total nitrogen (g).
2.2.16. Calculated protein efficiency ratio (C-PER)
C-PERs were calculated using the procedure suggested by Satterlee, Marshall, and Tension (1979) and
summarized by AOAC (1998). This procedure is based
on utilizing of the IVPD and the EAAs composition of
the different ours (untreated QPM our, QPM tempeh
our).
2.2.17. Protein digestibility corrected amino acid score
(PDCAAS)
The PDCAAS has been adopted as a current concept
in protein quality evaluation since it is more relevant to
human requirements (Sarwar & McDonough, 1990).
The PDCAAS method was conducted in two steps. The
rst step involved the determination of the TD (%) of
casein and QPM ours diets. In the second step, the
amino acid content was used to calculate the chemical
score for the protein of the diets. PDCAAS was
calculated according to the following equation:
PDCAAS (TD)(lowest AA score).
2.2.18. Statistical analysis
Results were analysed by the Desing Expert (2002)
using one-way analysis of variance (ANOVA) followed
by Duncans multiple range test comparisons among
means. Signicance was dened at Pp0:05.

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3. Results and discussion


3.1. Physico-chemical properties of QPM tempeh flour
Table 1 presents some physico-chemical properties of
QPM tempeh our. QPM tempeh our had lower
(Pp0:05) Hunter L value and higher (Pp0:05) DE
than the untreated QPM our (Table 1). Soaking and
cooking produced a signicant increase in Hunter L
value of QPM, meaning a lighter colour (data not
shown). Fermentation tended to lead towards a slightly
darker colour, probably due to the inuence of mycelia
colour and the drying step. Fermentation led to a DE
higher than the unfermented sample. However, colour
of QPM tempeh our appeared agreeable, but no
sensory studies were performed. The PSI is a measurement of our neness: higher PSI values mean smaller
our particles. The QPM ours showed PSI values from
71.94% to 79.59%; higher values corresponded to QPM
tempeh our. The bulk density measurements varied
from 0.492 to 0.544 g/cm3; QPM tempeh our showed
lower bulk density than QPM raw our. The range of
water activity for QPM ours corresponds to those
values where development of enzymatic activities,
growth of microorganisms and chemical reactions carry
out slowly, meaning a long shelf-life.
Gelatinization temperature and enthalpy were different (Pp0:05) between the untreated QPM and QPM
tempeh ours (Table 1). Gelatinization temperature was
higher for the QPM tempeh our than for the untreated
our. Gelatinization enthalpy was highest for the
untreated QPM our. Low gelatinization enthalpy value

Table 1
Physico-chemical properties of quality protein maize tempeh ours
Property1

Colour
Hunter L value
DE
PSI (%)
Bulk density (g/cm3)
aW
Tg (1C)
DH (J/g)
Total starch (g/100 g d.m.)
Resistant starch (g/100 g d.m.)
pH
WAI (g gel/g solids, d.m.)
WSI (g solids/g original solids)
Dispersability (%)

QPM Flour2
Unfermented

Tempeh

91.970.4a
9.270.5b
71.9471.87b
0.54470.01a
0.4970.03a
73.970.3b
2.7470.3a
62.971.9a
1.970.13b
6.1270.09a
1.370.02b
9.370.27b
34.470.49b

82.370.5b
21.770.6a
79.5972.22a
0.49270.01b
0.4870.05a
81.770.7a
1.9470.8b
56.972.5b
4.370.44a
4.570.10b
2.970.0a
8.270.21a
45.370.01a

1
DE total colour diference, PSI particle size index, aW water
activity, Tg gelatinization temperature, DH gelatinization enthalpy, WAI water absorption index, WSI water solubility index.
2
Means were separated by rows using Duncan0 s multiple range test.
Means with same letter are not signicantly different at Pp0:05.

in the QPM tempeh our suggests more severe processing conditions, because a drastic thermal treatment
produces starch gelatinization with a higher degree of
disorganization (Bello-Perez, Osorio-D az, Agama-Acevedo, Nunez-Santiago, & Paredes-Lopez, 2002) for this
reason, when QPM tempeh our was treated in the
DSC, only a small endotherm was found. Furthermore,
the lower enthalpy value in QPM tempeh our indicate
that, as a result of processing conditions, starch becomes
more gelatinized in QPM tempeh our than in raw QPM
our.
Appreciable variation (62.6 vs 56.9 g/100 g dry our)
for TS was recorded between untreated QPM and QPM
tempeh ours content (Table 1). TS values were slightly
greater in the untreated QPM our than in the QPM
tempeh our. This may be a consequence of partial
removal of nonstarch constituents during SSF process.
The RS value in the raw QPM our was similar to that
reported by Garc a-Alonso et al. (1999) and RendonVillalobos, Bello-Perez, Osorio-D az, Tovar, and Paredes-Lopez (2002) in maize our (Table 1). QPM
tempeh our showed signicantly higher RS values than
original raw QPM our. This result can be explained
based on the heat treatments that the grain suffers
during the SSF process. These treatments promoted the
interaction of the starch with other components
(proteins, lipids or itself) making it less accessible to
enzyme hydrolysis (Saura-Calixto et al., 1993). Biliaderis
(1992) reported that during the thermal processing of
foods rich in starch, RS is formed due to amylose
retrogradation. RSs have been introduced in recent
years as functional ingredients important to human
nutrition. The physiological importance of RS has been
investigated in relation to reduction of the glycemic and
insulinemic response to a food, as well as hypocholesterolemic effects and protective effects against colorectal
cancer (Asp, Van Amelsvoort, & Hautvast, 1996).
Fermented sample showed higher WAI than unfermented QPM; partial protein denaturation and starch
gelatinization, which occurred during cooking step,
could all be responsible for the increased WAI of
tempeh our. WAI of tempeh our might be related to
our starch damage. Heat processing of bean ours
resulted in higher water absorption capacities than
unfermented ours (Narayana & Narasinga, 1982).
3.2. Eaa content of QPM tempeh flour
EAAs content of unfermented QPM and tempeh
ours is shown in Table 2. Unfermented QPM and
tempeh ours contained 41.31 and 48.64 g EAA/100 g
protein, respectively; these values are higher than those
recommended by FAO /WHO for children 25 years old
(33.9 g EAA/100 g protein). When compared with FAO/
WHO reference standards, the proteins from unfermented QPM showed higher values of EAAs such as His,

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Table 2
Essential amino acid contents of QPM tempeh ours
EAA

His
Ile
Leu
Lys
Meth+Cys
Phe+Tyr
Thr
Trp
Val
Total
First lim EAA
Second lim EAA

QPM our1,2

EAA3 requirements 25 years

Unfermented

Tempeh

3.22b (1.69)
2.33b (0.83)
8.47b (1.28)
4.20b (0.72)
5.63b (2.25)
7.05b (1.11)
3.52b (1.04)
0.80b (0.73)
6.09a (1.74)
41.31
Lys
Trp

4.03a (2.12)
2.85a (1.02)
9.93a (1.50)
5.67a (0.98)
6.18a (2.47)
10.5a (1.67)
4.31a (1.27)
0.92a (0.84)
4.25b (1.21)
48.64
Trp
Lys

1.9
2.8
6.6
5.8
2.5
6.3
3.4
1.1
3.5
33.9

Means were separated by rows using Duncan0 s multiple range test. Means with same letter are not signicantly different at Pp0:05.
Values in parentheses are the essential amino acids score.
3
FAO/WHO (1991).
2

Leu, total sulphur (Met & Cys) amino acids, total


aromatic (Tyr & Phe) amino acids, Thr and Val;
however, these untreated proteins have lower levels of
Ile, Lys and Trp. In general, the EAAs content of
proteins from unfermented QPM was improved by SSF
process; the content of His, Ile, and Leu increased
signicantly (Pp0:05) in 0.81, 0.52, and 1.46 g/100 g
protein, respectively. Furthermore, total sulphur (Met &
Cys) and total aromatic (Tyr & Phe) amino acids
increased (Pp0:05) in 0.55 and 3.45 g/100 g protein,
respectively. However, Val levels decreased from 6.09 to
4.25 g/100 g protein. The most signicant effect of SSF
process on QPM proteins was the improvements in the
Lys and Trp contents; these EAAs had increments in
1.47 and 0.12 g/100 g protein, respectively. Other researchers (Addo, Lykins, & Cotton, 1996; Hamad &
Fields, 1979) have reported increments in Lys levels
when comparing fermented against unfermented maize.
Paredes-Lopez and Harry (1988) reported that among
the amino acid released during fermentation, the highest
values were for Lys and Met. They suggested that a
biochemical mechanism, such as transamination, might
be taking place during SSF.
The EAAs scores of proteins from unfermented and
fermented QPM our were evaluated taken into account
the suggested pattern of amino acid requirements for
children 25 years old (FAO/WHO, 1991) (Table 2). In
proteins from untreated QPM, Lys and Trp were the
rst and second limiting EAAs, with an EAAs score of
0.72. Sproule, Serna-Saldivar, Buckholt, Rooney, and
Knabe (1988), and Barragan-Salgado and Serna-Sald var (2000) reported EEA scores for proteins from
unfermented QPM of 0.72 and 0.66, respectively.
Therefore, the EEA scores and limiting amino acids of
raw QPM our were affected by the SSF process; in
proteins from QPM tempeh our, Trp and Lys were the

rst and second limiting EAAs, with EAAs scores of


0.84.
3.3. Nutritional properties of QPM tempeh flour
IVPD and biological values of proteins from unfermented and fermented QPM our are shown in Table
3. The IVPD was improved by SSF process; proteins
from unfermented QPM and QPM tempeh ours had
IVPD of 72.283.2%, respectively. Paredes-Lopez and
Harry (1989) reported an increase in IVPD of common
beans as a consequence of the same process. Increases in
IVPD could be explained by elimination of antinutritional factors (e.g. hydrolysis of phytic acid during
fermentation) and protein denaturation in the cooking
step, which result in proteins that are more vulnerable to
enzyme action (Mugula, 1992; Paredes-Lopez & Harry,
1989). The TD is an indicator of the amount of
nitrogen/protein absorbed from a particular diet. The
results of the animal studies showed that the TD of
QPM tempeh our increased signicantly (Pp0:05)
when compared with unfermented QPM our. As
expected, rats fed with the control casein diet used
dietary protein more efciently when compared to
counterparts fed with the untreated QPM and QPM
tempeh our (Table 3). The improved EAA patterns or
scores and the higher nitrogen retention values observed
in the QPM tempeh our (Table 2) clearly improved rat
performance (Table 3). PER and NPR were improved
(Pp0:05) as a consequence of the SSF. Wang, Ruttle,
and Hesseltine (1969) studied the effect of SSF process
with R. oligosporus, on protein quality of wheat by rats
assay methods; the PER of wheat was increased by
fermentation, this improvement was partially attributed
to increases in availability of lysine in wheat by SSF.
The QPM tempeh our had 89.6% of the PER value

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E.O. Cuevas-Rodrguez et al. / LWT 39 (2006) 10721079

Table 3
Nutritional properties of quality protein maize tempeh ours
Property1

Protein digestibility (%)


In vitro
In vivo
Apparent
True
PER
NPR
C-PER
PDCAAS

QPM Flour2

Casein

Unfermented

Tempeh

78.5071.2c

83.6070.9b

90.0071.2a

73.9071.6c
76.6071.1c
1.7870.05c
2.1370.02c
1.4370.8c
0.5570.03c

84.2072.1b
86.8070.8b
2.1070.03b
2.5670.03b
1.7470.6b
0.8370.02b

90.7071.7a
93.2071.1a
2.5070.05a
2.8670.05a
2.5070.7a
1.1170.01a

PER protein efciency ratio, NPR net protein retention, C-PER calculated protein efciency ratio, PDCAAS protein digestibility
corrected amino acid score.
2
Means were separated by rows using Duncan0 s multiple range test. Means with same letter are not signicantly different at Pp0:05.

found for the casein diet indicating that high Lys in


QPM tempeh our can be very benecial for groups of
people who depended on maize as the staple food. The
high quality of the protein of QPM tempeh our makes
these products particularly attractive for countries where
the high prevalence of protein-energy malnutrition is due
largely to the poor nutritional quality of the diet.
C-PERs for proteins from unfermented and fermented QPM ours were 1.43 and 1.54, respectively (Table
3); these values are higher than those reported by other
researchers (Hsu, Sutton, Banjo, Satterlee, & Kendrick,
1978; Sullivan & Carpenter, 1993): corn meal (1.1),
wheat our (0.8), soy our (1.3). C-PER model is based
on EAA prole and protein digestibility analysis (Hsu
et al., 1978; Sullivan & Carpenter, 1993).
PDCAAS is adopted as a current concept in protein
quality evaluation methodology since it is more relevant
to human requirements (Henley & Kuster, 1994; Sarwar
& McDonough, 1990). It is considered that PDCAAS
may replace the need for assessing the overall protein
quality. SSF process increased (Pp0:05) the PDCAAS
of untreated QPM our; PDCAAS of unfermented
QPM and QPM tempeh our were 0.55 and 0.73,
respectively. Both values are higher than those reported
for whole wheat (0.40), wheat gluten (0.25), oat (0.5),
and peanut meal (0.52). Moreover, Barragan-Salgado
and Serna-Sald var (2000) reported PDCAAS of 0.44
and 0.59 for weaning food produced from QPM and
regular maize, respectively.

4. Conclusions
The study showed that the SSF process could be
applied to improve the nutritional characteristics of
QPM. In comparison to unfermented QPM ours, the
QPM tempeh our showed higher (Pp0:05) gelatinization temperature, and RS, and a lower (Pp0:05)

gelatinization enthalpy, and TS. The contents of His,


Ile, Leu, Lys, Trp, Cys+Met and Tyr+Phe increased
(Pp0:05) as a consequence of SSF process. In unfermented QPM our the rst and second limiting EAAs
were Lys and Trp, respectively; QPM tempeh our had
as the rst and second limiting EAAs Trp and Lys,
respectively. The fermentation process improved
(Pp0:05) in vivo protein digestibility, PER, NPR,
calculated PER, and PDCAAS. Based mainly on its
nutritive value, fermented our may be considered for
the fortication of widely consumed cereal-based food
product (tortillas, bread, cookies, and atoles).
Acknowledgements
This research was supported by the Consejo Nacional
de Ciencia y Tecnolog a (CONACyT) Mexico, Consejo
Estatal de Ciencia y Tecnolog a del Estado de Sinaloa
(CECyT) and the Programmes Programa Integral para
el Fortalecimiento Institucional (PIFI, 2003) and
Programa Integral para el Fortalecimiento del Postgrado (PIFOP, 2003).

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