Food Chemistry, Vol. 63, No. 1, pp.

71-78, 1998
0 1998 Elsevier Science Ltd. All rights reserved
Printed in Great Britain
PII: SO308-8146(97)00199-4 030%8146/98 $19.00+0.00
ELSEVIER

Influence of enzymatic treatment on the

nutritional and functional properties of pea flour

Ma Jesh Periago,“” Ma Luisa Vidal,” Gaspar Ros,= Francisco Rinc6n,b Carmen Martinez,”
Gin& Lbpez,” Joaquin Rodrigo” & Isabel Martinez”
aDepartment of Food Science and Nutrition, Veterinary Faculty, Murcia University, 30071 Murcia, Spain
bDepartment of Food Science and Technology, Faculty of Veterinary, Cbrdoba University, 14014 Cdrdoba, Spain

(Received 26 March 1997; revised version received and accepted 8 September 1997)

The effect of enzymatic treatment on the nutritional value and functional prop-
erties of pea flour was investigated. Pea flour was hydrolyzed with acid protease
from Aspergillus saitoi, to give two different hydrolyzed pea flours. This enzy-
matic treatment led to a significant (p < 0.05) decrease in crude and true protein
and to an increase of free amino acids and non-protein nitrogen. The nutritional
value decreased, but an increase in the avilability of protein was expected as
result of lower trypsin inhibitor activity and phytic acid content in hydrolyzed
pea flours. The amino acid profile of unhydrolyzed pea flour was slightly modified
after enzymatic hydrolysis, increasing (significantly) the isoleucine, leucine, lysine,
cystine, phenylalanine, threonine, alanine, arginine and aspartic acid contents as
a result of the added enzyme. In addition, enzymatic treatment released hydro-
phobic amino acids, which significantly improved the protein solubility at acid
pH, the oil absorption capacity and the emulsification capacity of pea flours.
Protein solubility, foaming capacity, foam stability, water absorption capacity,
gelation capacity and green colour decreased. It was thus confirmed that treatment
with acid protease improves some functional properties of pea flour, but the effect on
nutritional properties was unclear. 0 1998 Elsevier Science Ltd. All rights reserved

INTRODUCTION proteins are hydrolyzed for many reasons, which range

from the improvement of nutritional and functional
Proteins are an essential food component because they properties, texture characteristics and for removal of
are the source of amino acids, needed for growth and odour, flavour, and toxic or antinutritive components
maintenance, and provide functional properties to foods (Lahl and Grindstaff, 1989). The most commonly used
(Giese, 1994). Commercially available protein foods are proteins in hydrolysis treatments are casein, whey and
obtained from a range of animal and plant sources and soya proteins (Lahl and Braun, 1994), but other protein
are used as functional ingredients (Amiot and Brisson, sources, such as legumes, have also been used success-
1985). Pea flour and pea protein isolates are examples fully (Amiot and Brisson, 1985).
of protein foods. Pea protein isolates are functional Pea flour, obtained from milled seeds, is a good
ingredients in terms of water and fat binding, emulsifi- source of protein (around 30% of total composition)
cation, and foaming and gelling characteristics (Giese, (Periago et al., 1996~). It also has high levels of non-
1994). They have been used to formulate non-dairy starch polyssacharides (or dietary fibre) and resistant
frozen desserts (Chan et al., 1992) and to replace the starch (Periago et al., 1994, 19966) and a high iron
albumen in sponge cakes (Giese, 1994). content (Periago et al., 1996~). However, as in the case
Over the last twenty years, the use of enzymes in the of other seed legumes, antinutritive factors such as
food processing industry has expanded rapidly (Faerge- phytic acid and trypsin inhibitor as well as their colour
man, 1994). For special foods, such as those destined and flavour, can limit the use of pea flour as an ingre-
for children, old people or athletes, protein food has dient in bakery products (Nielsen et al., 1980; Repetsky
been hydrolyzed (Gottschick, 1994). In general, food and Klein, 1982), meat products and snack feedstuffs
(Owusu-Ansah and McCurdy, 1991). For these reasons,
grain legumes were treated enzymatically to improve the
*To whom correspondence should be addressed. Fax: 0034 68 nutritional value of the protein (Lopez-Hemandez et al.,
364147; e-mail: mjperi@fcu.um.es 1977), to remove their beany flavour (Fujimaki et al.,

71
72 Ma J. Periago et al.

1968) and to reduce the content of antinutritive factors Treatment 1

such as polyphenols (Feldman and Vinnikova, 1973), The supension of pea flour was centrifuged at 3500 rpm
phytic acid (Morehouse and Malzahn, 1976; Li et al., for 10min. The superntant was discarded and the pellet
1989) and trypsin inhibitor (Li et al., 1989). was freeze-dried to obtain the hydrolyzed pea flour.
The aims of the present study were to study the chem-
ical and nutritional properties of pea flour protein after Treatment 2
enzymatic treatment, to evaluate the effect of enzymatic The suspension of pea flour was directly frozen and then
hydrolysis on the functional properties of pea flour and freeze-dried to obtain the hydrolyzed flour.
to explore the possible uses of the hydrolysed flour to
fortify some foodstuffs. Chemical analysis

Total nitrogen and crude protein (Nx6.25) were deter-

MATERIALS AND METHODS mined according to the micro-Kjedahl method (AOAC,
1990). True nitrogen and true protein was analysed fol-
Samples lowing the TCA precipitation method described by
Awolumate (1983). Non-protein nitrogen was calcu-
Wrinkle pea seeds (Pisum sativum, L.) (cultivar War- lated as the difference between total and true nitrogen
indo with a seed diameter from 8.3 to 8.8mm) were contents as recommended by Periago et al. (1996~). Free
selected for this study. They were harvested mechani- amino acids were determined with the ninhydrin reagent
cally and the pods removed by a shelling machine. (N- 1632, Sigma) compared with an amino acid standard
The peas were washed and the whole seeds were frozen (A-9656, Sigma) and measured at 580 nm using a double
and dried in a Virtis Freezer-drier model 10234 (Gardi- beam molecular spectrophotometer Hitachi model
ner, NY, USA) for 48 h. The dry pea seeds were milled U-2000 (Hitachi Ltd, Tokyo, Japan).
in a Moulinex Coffee-grinder (Alegon, France) with a
stainless-steel blade and passed through a US standard In vitro protein digestibility
40 mesh sieve.
In vitro protein digestibility was determined by the
Enzymatic hydrolysis multienzymatic technique (Satterlee et al., 1982) and
was calculated from the change in pH of a sample
Enzymatic treatment of the pea flour protein involved digested within a 20min period with a mixture of the
acid protease from Aspergillus saitoi, commercially following enzymes: porcine pancreatic trypsin (Type IX,
called ‘Molsin’ (P-2143, Sigma Chemical, St Louis, MO, T-01 34, Sigma), bovine pancreatic ar-chymotrypsin
USA). Trial experiments were performed to determine (Type x11, C-4129, Sigma), pepsin from porcine sto-
the best time, temperature and ratio of enzyme to sub- mach mucose (Grade I, P-6887, Sigma) and bacterial
strate using: temperatures of 40, 60 and 90°C times of protease (Pronase E, P-5147, Sigma). Sodium caseinate
90, 120 and 180 min, and enzyme to substrate ratios of (Sigma) was used as reference material.
0.27, 0.80, 1.62, 2.70, 5 and 10%. Enzyme activity was
measured as the amount of solubilized protein in a Amino acid profiles
3.3% trichloroacetic acid solution and as the free amino
acids released after hydrolysis, both quantified In order to prepare the sample, the pea flour protein
spectrophotometrically using Lowry’s technique at was digested by acid hydrolysis as described by Satterlee
71Onm (Lowry et al., 1951) and the ninhidrin method at et al. (1982). The amino acid composition was deter-
580nm (Awolumate, 1983) respectively. As a result of mined with an amino acid analyser LKB Alpha Plus
these trials the following parameters were selected as the (Parmacia LKB Ciochrom Ltd, Cambridge, England),
best treatment conditions: temperature 40°C time comparing the chromatogram of the samples with a
90min and an enzyme/substrate ratio of 1:lO. These standard solution (Part. No. 40 00 9037, Pharmacia
conditions led to maximum hydrolysis of the protein in LKB Biochrom Ltd). The tryptophan content was ana-
pea flour. Prior to enzymatic treatment, a suspension of lysed by a calorimetric technique (Sastry and Tummuru,
pea flour was prepared by adding 100ml of distilled 1985) after alkali hydrolysis of the pea flour protein
water to 4g of flour. The suspensions were acidified to with 5 M NaOH.
pH 2.8 with 3 M HCl solution, and placed in a water
bath and the enzyme was added. The enzymatic Trypsin inhibitor activity assay
hydrolysis was conducted at 40°C for 90min. The reac-
tion was stopped by increasing the pH to 6.0 with 2N Trypsin inhibitor activity was determined using the
NaOH solution, and the enzyme was inactivated by method of Kakade et al. (1974), modified by della Gatta
placing the samples in a boiling water bath for 15 min. et al. (1988). This procedure measures the inhibition by
The resulting hydrolyzed solution was treated in two aliquots of pea flour extract, of the enzyme activity
differents ways: of bovine trypsin (T-8003, Sigma) on the synthetic
 Influence of enzymatic treatment on the nutritional andfunctional properties of pea pour 73

substrate DL-benzoyl arginine p-nitroanilide (DL- added to 6g of sample in a cylinder volumetric flask,
BAPNA, B-4875, Sigma), expressing the results as and blended with a speed of between 7000 and 8000 rpm
trypsin inhibitor activity units (TIU). One TIU is with an Omnimixer homogenizer. Foam stability was
defined as a decrease in absorbance of the test solution measured as the percentage increase in volume as
at 410 nm by 0.01 units in 10min. A sample blank recorded before and after blending (Lin and Humbert,
without enzyme was analysed for each sample to correct 1974). To study the foaming stability, the volume of the
for any residual turbidity or interaction between sub- mixtures was recorded as function of time over a period
strate and a sample solution. from 5 to 120min (Lin and Humbert, 1974).

Phytic acid Gelation capacity

Gelation capacity was determined with different pea
Phytic acid was extracted from pea flours with 3% flour suspensions, using the following flour/water ratios:
H2S04 solution, and precipitated as phytate-ferric com- 2, 4, 6, 8, 10, 12, 14, 16% (w/w). 5ml of these suspen-
plex, which was converted to ferric hydroxide by adding sions were introduced into a test tube and placed in a
1.5 M NaOH solution. After boiling, the phytic acid was boiling water bath for 1 h, followed by rapid cooling
released as soluble sodium phytate, which was measured under cold running tap water. The tubes were further
as phosphorus using an Inductively-Coupled Plasma cooled for 2 h at 4°C. The gelation capacity was taken
Atomic Emission Spectrometer (ICP-AES) model to be the concentration which prevented the sample
JY 70 Plus (Jobin Yvon, Paris, France) (Plaami and from slipping when the test tube was inverted (Coffman
Kumpulainen, 1991). and Garcia, 1977).

Functional properties Osmolality

The osmotic pressure, measured by freezing point
Protein solubility depression, was determined in a suspension of 1 g of pea
Protein solubility, as a function of pH, was determined flour in lOm1 of distilled water using a Micro-Osmo-
by extraction of the protein at different pH values and metre model 3 MO-Plus (Advanced Instruments Inc.
subsequent determination of the protein in the extract Massachussets, USA).
using the Lowry’s calorimetric method, as described by
Sathe and Salunke (1981). Colour
Colour was determined according to the ‘L’ (luminos-
Water and oil absorption capacities ity), ‘a’ (greeness) and ‘b’ (yellowness) values using a
The water absorption and oil absorption capacities were calorimeter Minolta Chroma Meter II Reflectance CR-
carried out following the procedure described by Beu- 2000 (Minolta Limited, Milton Keynes, UK).
chat (1977). One gram of pea flour was mixed thor-
oughly with 10 ml of distilled water or sunflower oil in a Statistical analyses
volumetric test tube, and then centrifuged at 55OOOxg The statistical analyses of the data were performed with
for 30min. The water absorption and oil absorption a SYSTAT program version 5.0 (Wilkinson and Howe,
capacities were calculated as grams of water or oil 1992). Results were expressed as the mean values f
absorbed per gram of flour, respectively, considering a standard deviation of three separate determinations. To
density of 1 gml-’ for water and 0.9166gmlli for sun- ascertain the significance among means of the samples,
flower oil. Tukey’s means separation test was applied. Unless
otherwise stated, ~~0.05 was used to establish sig-
Emulsification capacity nificant differences.
The emulsification capacity of pea flour was determined
by the method of Beuchat (1977). Two grams of pea
flour were mixed with lOOm1 of distilled water and RESULTS AND DISCUSSION
blended at low speed (1200rpm) for 30s at 25°C using
an Omnimixer homogenizer (Omni International, Effects of the enzymatic treatment on the chemical and
Waterbury, CT, USA). An aliquot of 5 ml was taken and nutritional composition of pea flour
sunflower oil was added from a burette at a constant rate
of 5 ml min-’ with continuous blending, until the break- Table 1 shows the chemical and nutritive parameters in
point (indicated by separation of the oil from the aqu- unhydrolyzed pea flour and in both hydrolyzed pea
eous phase) was reached. The emulsification capacity flours. In hydrolyzed pea flour, the enzymatic treatment
was expressed as ml of oil emulsified per g of protein. led to a significant @ < 0.05) reduction in the total and
protein nitrogen, and crude and true protein contents,
Foaming capacity and foam stability whereas the non-protein nitrogen and free amino acid
To ascertain the foaming capacity in non-hydrolyzed contents increased significantly (p < 0.05). The content
and hydrolyzed pea flour, 200 ml of distilled water were of crude protein was 26.1 g 100 g-’ in unhydrolyzed pea
74 Ma J. Periago et al.

Table 1. Effects of enzymatic treatments on the nutritional value of protein and on the content of antinutritive factors of pea flour’

Hydrolyzed pea flours

Parameters Unhydrolyzed pea flour Treatment 1 Treatment 2

Total nitrogen (%) 4.18&0.05” 3.18h0.1Sb 3.59 zt 0.29b

Crude protein (%) 26.1 f 0.29a 19.9 f 0.3oc 22.4 zk1.80b
Protein nitrogen (%) 3.12k0.31p 1.07h0.12b 0.92 zt 0.22b
True protein (%) 19.5 f 0.430 6.65 f 0.7@ 5.75 f 1.43b
Non-protein nitrogen (%) 1.oo zt 0.02b 2.05hO.14” 2.50 zt 0.35”
Free aminoacids (mg g-l) 0.28 f 0.01’ 4.45 f 0.12” 6.72iO.12”
In vitro protein digestibility (%) 82.3 f 0.55” 71.1 It 1.436 72.9 f 0.67’
Trypsin inhibitor (TIA mg-‘) 4.72 f 1.08O 2.06 zt 0.37b 2.11 f 0.52b
Phytic acid (mg g-i) 4.35 f 0.050 1.49 f 0.336 1.73 f 0.356

l Mean f standard deviation of three determinations exnressed as dry weight. Different letters within the same row are significantly
different at p < 0.05.

flour, whereas in hydrolyzed flours, the crude protein nificantly (p < 0.05) from 4.72 to 2.06 TIU mg-’ and the
content decreased with decreases in the total nitrogen latter from 4.35 to 1.49 mg g-t. This reduction in trypsin
content. In general, the crude protein content in peas inhibitor activity might be related to heating during the
varies widely as a result of the variety, size, and genetic enzymatic treatment, or due to the action of the enzyme
and environmental factors (Savage and Deo, 1989; Ros on the protein, leading to denaturation of the protein
and Rincon, 1990; Periago et al., 1996a). The variability chains (Vidal et al., 1995), whereas the reduction in the
observed in the crude protein content was also observed phytic acid content was mainly attributed to the release
in the true protein content. In peas, true protein is made of phosphorus as orthophospbate (Morehouse and
up of 65-80% globulin and 2&35% albumin (Owusu- Malzahn, 1976), probably due to the activation of the
Ansah and McCurdy, 1991), increasing with pea size endogenous phytase. A low antinutritive factor content
due to the protein synthesis which takes place in the has an important effect on the nutritional protein value,
seed kernel during development of the plant, in order to because trypsin inhibitor and phytic acid significantly
build up a reserve of protein ready for germination reduce the in vitro protein digestibility (Carnovale et al.,
(Periago et al., 1996a). The true protein content 1988; Al-Wesali et al., 1995). Moreover, a lower phytic
decreased markedly after enzymatic treatment of the acid content has an important effect on mineral bioa-
pea flour. This effect is mainly due to hydrolysis of the vailability, since this compound forms an insoluble
pea protein, since the enzymatic treatment releases pep- complex with divalent cations like zinc, copper, iron,
tides and free amino acids from protein, increasing the manganese and calcium, thus reducing bioavailability
non-protein nitrogen, which might be solubilized in the (Harland and Oberleas, 1987).
NaOH 0.2% and cannot be precipitated by TCA. The amino acid composition of pea flours before and
In vitro protein digestibility decreased significantly in after enzymatic treatment are shown in Table 2. In
hydrolyzed flour, from 82.3% to 71 .l %, probably due general, the most abundant amino acids were glutamic
to the fact that the remaining proteins are more resis- acid, aspartic acid, lysine, and leucine, whereas the sul-
tant to the enzymes hydrolysis. However, Lbpez-Her- phur amino acid content (cystine and methionine) was
nandez et al. (1977) have reported that hydrolyzed low compared with that of other protein sources, such
protein shows better availability since low molecular as have been described by several authors (Holt and
weight peptides and amino acids are released. These are Sosulsky, 1979; Lee et al., 1982; Sosulski and McCurdy,
readily absorbed and available to the human body and 1987; Savage and Deo, 1989; Periago et al., 1996a).
could easily fulfil the daily quantities of protein recom- However, the amino acid content of peas is known to be
mended for special groups that require dietetic control affected by cultivar, growing season, and size (Periago et
(Frrakjaer, 1994). Higher in vivo protein digestibility al., 1996a). There were no significant differences in the
values should therefore be expected in hydrolyzed pea histidine, methionine, tyrosine, valine, triptophan,
flours, since there is a marked increase in free amino glutamic acid, proline and serine contents between
acids after enzymatic hydrolysis_ The free amino acid unhydrolyzed and hydrolyzed pea flours, whereas iso-
content was significantly @ < 0.05) higher in the hydro- leucine, leucine, lysine, cystine, phenylalanine, threo-
lyzed pea flour obtained with treatment 2, because, in nine, alanine, arginine and aspartic acid increased
treatment 1, the supernatant resulting from hydrolysis significantly (p < 0.05) after enzymatic treatment of pea
was discarded, which meant the solubilized free amino flour. The increase in hydrophobic amino acids such as
acids were removed from the pea flour. isoleucine, leucine and lysine is important, due to the
The enzymatic treatment with acid protease led to a effects that these have on the physical and functional
considerable reduction in the trypsin inhibitor and phy- properties of food proteins (Giese, 1994; Mahmoud,
tic acid contents, the former’s activity decreasing sig- 1994). The hydrolyzed pea flours supplied a higher
 Influence of enzymatic treatment on the nutritional andfunctional properties of pea flour 75

Table 2. Effect of enzymatic treatment on the amino acid profiles (g 100 g-’ of protein) of pea flour’
Hydrolyzed pea flours FA02 ‘Ideal’ Human requirements
Amino acid Unhydrolyzed pea flour Treatment 1 Treatment 2
Essential
His 0.96 f 0.13” 1.11 ZkO.28” 1.00*0.1OQ - 16
Ile 2.75 f 0.2gb 3.53%0.15” 3.85kO.14” 4 13
Leu 4.76 f 0.36b 6.74*0.19” 6.20~0.21” 7 19
Lys 4.3 1 •k 0.42b 3.99 f 0.69* 5.66ItO.17” 5.5 16
Met 0.83*0.15” 1.03~0.19” 1.04 f 0.04” 3.5 17*
Cys 0.32 * 0.04b 0.77*0.11= 0.58rtO.15”b - 17*
Phe 2.53f0.21b 3.30*0.13= 3.10*0.14= 19+*
Tyr 1.55%0.15” 1.38~kO.51” 1.86rt0.29n 6- 19**
Thr 2.64*0.196 3.62kO.17” 3.41 ZtO.11’ 4 9
Trp 1.03 f 0.08” 0.87~kO.10~ 1.02 f 0.04” 1 5
Val 2.98 i 0.54a 3.62kO.12” 3.78 i 0.45a 5 13
Non-essential
Ala 3.26* 0.41c 4.11*0.136 4.80 f 0.17” -
Arg 2.55 f 0.74b 3.51 f0.19Qb 3.97 rt 0.52” -
Asp 3.28 f 0.28” 4.77 f 0.21” 5.08rt0.15a - -
Glu 10.3 f 0.94” 10.5 f 1.91” 12.0 rt 0.23”
Gly 2.70 f 0.40b 3.25 f 0.05* 3.37 f 0.09” -
Pro 2.60~k0.21~ 3.11 f 0.47O 2.94 f 0.23” - -
Ser 3.00 Zk0.20” 3.46 & 0.75” 3.82~tO.13~ - -

‘Mean * standard deviation of three determinations. Different letters within the same row are significantly different at p < 0.05.
‘FAO ‘Ideal’, data from FAO/OMS (1973).
‘Human requirements, data from FAO/OMS (1992).
*Human requirements expressed as Met + Cys.
**Human requirements expressed as Phe + Tyr.

proportion of the amino acid requirements of human

than the non-hydrolyzed pea flour, and the quantities of
essential amino acids, isoleucine, leucine, lysine, and
threonine covered 90.5% to 100% of the FAO ‘ideal’
(FAO, 1973).

Effects of enzymatic treatment on the functional

properties of pea flour

The solubility profiles of protein from unhydrolyzed

and hydrolyzed pea flours are shown in Fig. 1. The
solubility of pea protein is low at acid pH, but increases
in more basic pH conditions (Owusu-Ansah and
McCurdy, 1991). The profile of unhydrolyzed pea flour 2 3 4 4‘5 5 5.5 6 7 8 9 10 11
showed a solubility curve with a broad minimum in the
pH range of 3-6. Below pH 3, the solubility increased
PH
reaching a maximum of 35%. Above pH 6 there was a
marked increase in solubility with a maximum of Fig. 1. Protein solubility curves at different pH of unhy-
42.11% at pH 11. Similar solubility patterns were drolyzed pea flour and hydrolyzed pea flours.
reported in pea protein (Megha and Grant, 1986;
Sosulski and McCurdy, 1987), and in other legume seed hydrolyzates’ hydrophilic&y (Mahmoud, 1994). The
proteins such as those recovered from cowpea flour additional heat treatment applied during enzymatic
(Abbey and Ibeh, 1988) and brown beans (Abbey and treatment might also cause a slight modification in the
Ibeh, 1987). Both hydrolyzed pea flours showed similar solubility of proteins from vegetal sources (Megha and
protein solubility values in the acid pH range. In general, Grant, 1986; Abbey and Ibeh, 1987, 1988; Prakash and
enzymatic hydrolysis modifies the solubility character- Ramanatham, 1995).
istics of all food proteins, not only those from vegetal The foaming capacities and the foam stabilities of
sources but also from animal sources (Frskjaer, 1994). protein from unhydrolyzed and hydrolyzed pea flour
The enhanced solubility of the hydrolyzates is due to are represented in Fig. 2. Unhydrolyzed pea flour
their smaller molecular size and the newly exposed showed a higher foam capacity, by developing high
ionizable amino and carboxyl groups, that increase the initial foam volumes and maintaining their relatively
76 Ma J. Periago et al.

is significantly correlated with the content of crude pro-

tein (r=0.87, ~~0.01) (Sosulski and McCurdy, 1987),
increasing as the protein content increases (Megha and
Grant, 1986). For this reason, the hydrolyzed protein of
treatment 2 showed the lowest water absorption capa-
city, because of its low true protein content (Table 1).
As regards oil absorption capacity, the hydrolyzed pea
flours showed a significant (p ~0.05) increase with
respect to that of the corresponding flour (Table 3),
suggesting that the protein composition of the fractions

lo;],, ] was the principal determining factor in the response to

these functional tests (Sosulski and McCurdy, 1987).
Since lipid binding depends on the surface availability
of hydrophobic amino acids, the increased oil absorp-
0 beaten 1 5 10 30 60 120 tion capacity could be attributed to an increase in these
amino acids during enzymatic treatment (Lahl and
Time (mid Braun, 1994), as mentioned earlier (Table 2). Therefore,
Fig. 2, Foaming capacity and foam stability of unhydrolyzed the heat treatment applied during enzymatic treatment
pea flour and hydrolyzed pea flours. could lead to some modifications of the pea protein,
since heat treatment of pea flour and pea concentrate
coarse structure throughout the 2 hour holding period. modified the globulin fraction and increased the oil
Hydrolyzed pea flour formed less foam on whipping absorption capacity (Megha and Grant, 1986).
than the corresponding pea flour, as result of the pro- The emulsification capacities of pea flour proteins are
tein’s hydrolysis and also probably due to the loss of shown in Table 3, the enzymatic treatment increasing
soluble low-molecular weight proteins during enzymatic this capacity significantly (pcO.05) from 35.85ml of
treatment (Sosulski and McCurdy, 1987). The hydro- sunflower oil g-l of sample in unhydrolyzed pea flour to
lyzed pea flour of treatment 1, showed a slightly higher 44.52ml of sunflower oil g-’ of sample in hydrolyzed
foam capacity immediately after whipping than the pea flour of treatment 1. It is generally recognised that
hydrolyzed pea flour of treatment 2, perhaps due to proteins are improved by enzymatic hydrolysis, and the
losses of low molecular weight protein. However, the emulsification capacity of soya protein isolate hydro-
foam stability of both hydrolyzed pea flours were simi- lyzed by fungal protease and whey protein and casein
lar, the foam volume only being maintained for 10 min hydrolyzed with trypsin increased after treatment
after whipping. (Haque and Mozaffar, 1992). However, the extent of
Table 3 shows the effect of the enzymatic treatment of hydrolysis could lead to a reduction in the emulsifica-
pea flour on some functional properties. The water tion capacity of proteins, probably due to exposure of
absorption capacity of the hydrolyzed pea flour from the hydrophobic protein interior, which would enhance
treatment 2, was significantly (~~0.05) different from adsorption at the interface forming a cohesive inter-
that of unhydrolyzed pea flour and hydrolyzed pea flour facial film, and the hydrophobic residues interacting
of treatment 1. The water uptake ranged from 1.5 to with oil and the hydrophilic residues interacting with
2.9 g of waterg-’ of sample. This functional property water (Mahmoud, 1994). The enzymatic treatment of
depends on the protein content but mainly on the phy- pea flour using acid protease under the conditions
sical interactions between water and protein (Cheftel et selected, led to a substantial enhancement of the emul-
al., 1989). The water absorption capacity in pea protein sifying capacity of the pea protein, which might have

Table 3. Effects of enzymatic treatment on the functional properties of pea flour’

Hydrolyzed pea flour

Functional properties Pea flour Treatment 1 Treatment 2
Water absorption capacity (g g-t) 2.9*0.10” 2.9zkO.10” 1.53 f 0.066
Oil absorption capacity (g gg ‘) 1.06~0.10c 1.77*0.10= 1.38+0.056
Emulsification capacity (ml gg ‘) 35.85 f 1.25b 44.52 z!z1.76” 42.37 f 2.05”
Gellation capacity (%) 8 14 12
Osmolality (m Osm Kg-’ HzO) 63.3 zk4.71’ 90+ 14.146 253*4.71a
Colour (coordenadas Lab)
L 70.93 f 0.80” 64.43 f 0.09b 55.78 f 1.30’
-a -11.77~0.18c + 19.81 ztO.05” + 17.14*0.226
b 26.20 z!z0.43b 37.22 f O.OOb 37.86* 0.23O
‘Mean f standard deviation of three determinations. Different letters within the same row are significantly different at p < 0.05.
 Influence of enzymatic treatment on the nutritional and functional properties of peaJlour 71

been due to the release of hydrophobic amino acids osmolality. However, a sensory study should be
(Table 2), thus increasing the interactions between oil developed to ascertain the effect of the enzyme on pea
and proteins. flour flavour.
The gelation concentrations for the unhydrolyzed and
hydrolyzed pea flours (treatments 1 and 2), were 8, 14
and 12%, respectively (Table 3). Protein hydrolyzates ACKNOWLEDGEMENTS
show a much reduced capacity to form gels, after
heating, than the corresponding intact proteins (Mah- Acknowledgements are made to the grants projects:
moud, 1994). This effect was also observed in the pea PIB 93/l 10 of the ‘Comunidad Autbnoma de la
flours studied. Unhydrolyzed flour had a higher gelation Regibn de Murcia’ and AL1 94-0338 of the Spanish
capacity than the hydrolyzed samples. Government (CYCIT), and to research grant from Hero
Osmolality is an important physical characteristic in Spain S.A.
hydrolyzed protein when it is used to prepare nutritional
formulas for both children and adults. A solution of
high osmolality may draw large quantities of water into REFERENCES
the small intestine, causing severe diarrhoea, possible
dehydration, and disruption of the electrolyte balance, Abbey, B. W. and Ibeh, G. 0. (1987) Functional-properties of
as well as inducing nausea, vomiting and abdominal raw and heat processed brown bean (Cunavaliu roseu DC)
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