FOCH 4202 No. of Pages 5, DTD = 5.0.

1
ARTICLE IN PRESS
29 July 2004; Disk Used

Food
Chemistry
Food Chemistry xxx (2004) xxx–xxx
www.elsevier.com/locate/foodchem

2 Effect of fermentation on the functional properties of sorghum flour

OF
a,*
3 Abd Elmoneim O. Elkhalifa , B. Schiffler b, R. Bernhardt b

a
4 School of Family Sciences, Ahfad University for Women, P.O. Box 167, Omdurman, Sudan
b
5 Universität des Saarlandes, FR 8.8 Biochemie, Saarbrücken, Germany

Received 1 March 2004; received in revised form 27 May 2004; accepted 27 May 2004

PRO
8 Abstract

9 Sorghum flour was fermented by the traditional Sudanese method of fermentation for 24 h, taking samples every 8 h, and selected
10 functional properties were studied. Results showed that fermentation increased the protein solubility of sorghum flour in the acidic
11 range (pH 2–4). Fermented sorghum flour had a least gelation concentration of 6% after 16 h of fermentation, while it was 18% for
12 unfermented sorghum. Fermentation also increased oil-binding capacity, emulsifying capacity and emulsifying stability, while it de-
13 creased the water-binding capacity. Sorghum flour, fermented or unfermented, showed no foam capacity.
14 Ó 2004 Published by Elsevier Ltd.
TED
15 Keywords: Sorghum flour; Fermentation; Functional properties
16

17 1. Introduction 1983; Elkhalifa & El Tinay, 1994, 1995; El Tinay, Abdel 34

Gadir, & El Hidai, 1979). After fermentation, in vitro 35
REC

18 Sorghum (Sorghum bicolor (L.) Moench) is a crop protein and starch digestibilities increased significantly, 36
19 that is widely grown all over the world for food and as well as available lysine, leucine, isoleucine and methi- 37
20 feed. It is one of the main staples for the worldÕs poorest onine, while tannin content and phytic acid decreased 38
21 and most insecure people. It is a key staple in many during fermentation of sorghum flour. 39
22 parts of the developing world, especially in the drier Celiac disease continues to be a major health problem 40
23 and more marginal areas of the semi-tropics. Various in many countries. As a result of this, various efforts are 41
24 processing methods are used for preparation of foods being made to solve this problem through the introduc- 42
25 from sorghum; among them, fermentation is a unique tion of methods for increased utilisation of less popular 43
26 method for food preparation in Sudan and in Africa foodstuffs. 44
OR

27 in general. Recently, there has been increased interest in sor- 45

28 Information is available on the predominant micro- ghum as a gluten-free cereal to substitute the gluten-rich 46
29 organisms involved in the fermentation of sorghum cereals in the diet of people suffering from celiac disease. 47
30 flour (Abdel Gadir & Mohamed, 1977; Hamad, Bocker, The functional properties of sorghum proteins can be 48
31 Vogel, & Hammes, 1992; Mohammed, Steenson, & Kir- used to define how flour proteins can be used to supple- 49
UNC

32 leis, 1991) and on its nutritive value (Axtell et al., 1981; ment or replace more toxic protein sources. There is, 50
33 Eggum, Monowar, Bach Knudsen, Munck, & Axtell, however, no information on the functional properties 51
of fermented sorghum flour and this information is es- 52
*
sential for determining potential uses of this product 53
Corresponding author. Tel.: +249-15-554870; fax: +249-15-
in food formulation. The aim of this study was to deter- 54
553363.
E-mail address: [email protected] (Abd Elmoneim O. Elkha- mine the functional properties of fermented and unfer- 55
lifa). mented sorghum flour. 56

0308-8146/$ - see front matter Ó 2004 Published by Elsevier Ltd.

doi:10.1016/j.foodchem.2004.05.058
 FOCH 4202 No. of Pages 5, DTD = 5.0.1
ARTICLE IN PRESS
29 July 2004; Disk Used

2 Abd Elmoneim O. Elkhalifa et al. / Food Chemistry xxx (2004) xxx–xxx

57 2. Materials and methods 2.6. Water- and oil-binding capacity 102

58 2.1. Materials Two grammes of each flour sample were weighed into 103
a pre-weighed centrifuge tube and 20 ml of distilled wa- 104
59 A low-tannin sorghum cultivar (Tabat), obtained ter were added. For oil binding, 20 ml sunflower oil were 105
60 from the Food Research Centre, Shambat, Sudan, was added. Samples were vortexed and allowed to stand for 106
61 used in this study. The seeds were carefully cleaned 30 min at 25 ± 2 °C before being centrifuged at 4000g 107
62 and ground in a hammer mill to pass through a 0.4 for 25 min. Excess water or oil was decanted by invert- 108
63 mm screen, and the flour was stored in polyethylene ing the tubes over absorbent paper and samples were al- 109
64 bags at 4 °C. lowed to drain. The weights of water and bound oil 110

OF
samples were determined by difference. 111
65 2.2. Fermentation of sorghum
2.7. Emulsifying activity and stability 112
66 Sorghum flour was fermented according to the tradi-
67 tional method practised by the Sudanese housewife, as The method of Yasumatsu et al. (1972) was used. 113

PRO
68 described by El Tinay, El Mehdi, and El Soubki Emulsions were prepared with 1g of each sample, 50 114
69 (1985). Fermentation was earned out at 37 °C for peri- ml of cold distilled water (4 °C) and 50 ml of sunflower 115
70 ods of 0, 8, 16, and 24 h. After a distinct incubation pe- oil. The flour samples were dispersed with a Waring 116
71 riod, the samples were dried in a hot air oven (Heraeus blender. Each blended sample was divided equally into 117
72 UT 5042, Germany) at 60 °C for 16 h. Dried samples 50 ml centrifuge tubes. One centrifuge tube was directly 118
73 were ground to pass a 0.4 mm screen and stored in poly- centrifuged at 4000g for 10 min while the other was cen- 119
74 ethylene bags at 4 °C prior to analysis. trifuged under the same conditions after heating in a wa- 120
ter bath at 80 °C for 30 min and cooling to room 121
75 2.3. Protein solubility temperature (25 °C). The height of the emulsified layer, 122
as a percentage of the total height of material in the un- 123
76
TED
Protein solubility was determined in the pH range 2– heated tubes, was used to calculate the emulsifying ac- 124
77 12 for both fermented and unfermented sorghum flour. tivity and stability, respectively, using the following 125
78 A one gramme sample was dispensed in 60 ml distilled formulas: 126
79 water and the pH was adjusted with NaOH or HCl us- height of emulsion layer
80 ing a pH meter (766 Calimatic/Germany). The disper- Emulsion activity ð%Þ ¼
100;
height of whole layer 128
81 sion was continuously stirred in an orbital shaker at
82 150 rpm for 2 h at 25 °C, and then centrifuged (Sigma Emulsion stability ð%Þ ¼
83 Laborzentrifugen, Osterode, Germany) at 2000g for 20
REC

height of emulsion layer after heating

84 min. The supernatant was collected and the soluble pro-
height of whole layer
85 tein was determined by the procedure of Lowry, Rosen-

100:
86 brough, Fair, and Randall (1951). The percentage of
130
87 soluble protein was calculated and plotted against the
88 corresponding pH values. Height was measured in centimetres using a transparent 131
graduated ruler. 132
89 2.4. Least gelation concentration
2.8. Foaming capacity 133
OR

90 The least gelation concentration was determined by

91 the method of Coffman and Garcia (1977) as modified The foam capacity was determined by the method of 134
92 by Akubor and Chukwu (1999). Using flour concentra- Narayana and Narsinga Rao (1982) using two grammes 135
93 tions of 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18% and of each sorghum flour sample. 136
94 30% (w/v).
2.9. Statistical analysis 137
UNC

95 2.5. Bulk density

Two separate batches, for a particular treatment, 138
96 Bulk density (BD) was determined by the method of were taken and analysed separately and the figures were 139
97 Wang and Kinsella (1976). Ten grammes of the tested then averaged. Although sample material was of one 140
98 flour were placed in a 25 ml graduated cylinder and season, it is not expected that significant variation in 141
99 packed by gentle tapping of the cylinder on a bench functional properties would occur among several sea- 142
100 top, ten times, from a height of 5–8 cm. The final volume sons. Data were assessed by analysis of variance (ANO- 143
101 of the test flour was measured and expressed as g/ml. VA) (Snedecor & Cochran, 1987) and by DuncanÕs 144
 FOCH 4202 No. of Pages 5, DTD = 5.0.1
ARTICLE IN PRESS
29 July 2004; Disk Used

Abd Elmoneim O. Elkhalifa et al. / Food Chemistry xxx (2004) xxx–xxx 3

145 multiple range test with a probability P 6 0.05 (Duncan, protein solubility was in the following order 8 178
146 1955). h > 16 > 24. According to Kinsella (1979), seed proteins, 179
which are soluble at pH 4–8, could be used in ‘‘vegetable 180
milk’’ beverages; this might also be considered for sor- 181
147 3. Results and discussion ghum. 182

148 3.1. Protein solubility profile 3.2. Least gelation concentration 183

149 Protein solubility characteristics are influenced by The ability of fermented and unfermented sorghum 184
150 factors such as origin, processing conditions, pH, ionic flour to form gel was measured. The least gelation con- 185

OF
151 strength and the presence of other ingredients (Kinsella, centration (marked as + in Table 1) for fermented sam- 186
152 1979). The protein solubility profiles at various pH-val- ples was 8, 6 and 6% for the three fermentation periods 187
153 ues of unfermented and fermented sorghum flour are 8, 16 and 24 h, respectively, while it was 18% for the un- 188
154 shown in Fig. 1. The unfermented sorghum flour had fermented sorghum flour (Table 1). The least gelation 189
155 minimum protein solubility at pH 4, while the minimum concentration reported for legume flours was 14% for 190

PRO
156 protein solubility for all fermented was samples was at lupin seed proteins (Sathe, Salunke, & Deshande, 191
157 pH 6. These results showed that fermentation was shift- 1982), and 6% for defatted sesame seeds (Inyang & 192
158 ing the solubility of sorghum proteins by 2 pH units. Nwadimkpa, 1992). Gelation is an aggregation of dena- 193
159 The protein of the fermented samples was more soluble tured molecules. Fermentation may have denatured the 194
160 at the isoelectric pH than was the control. This could be sorghum proteins and, thus, caused more aggregation 195
161 attributed to structural changes in the protein of the fer- than in the unfermented sorghum flour. These results 196
162 mented samples. It could also be due to inactivation of suggest that fermented sorghum flour would be a good 197
163 anti-nutritional factors (phytates) caused by the fermen- gel-forming or firming agent, and would be useful in 198
164 tation. Highest solubilities occurred at pH 12 and values food systems such as pudding and snacks which require 199
165 determined were 76.7%, 57.3%, 55.2% and 55.2% for un- thickening and gelling. 200
TED
166 fermented, 8-, 16-, 24-h fermented sorghum flour, re-
167 spectively. At pHs 2–4, fermented sorghum flour had 3.3. Bulk density 201
168 higher protein solubility than unfermented. The high
169 solubility at pHs 2–4 for fermented sorghum flour could Fermentation of sorghum flour for 24 h decreased the 202
170 be attributed to proteolytic activity in fermenting seeds, bulk density of the sorghum flour by about 10% 203
171 yielding peptides and free amino acids, which increase (Table 2). The decrease in bulk density of fermented 204
172 nitrogen solubility in water (Beuchat, 1976). It was ob- flour would be an advantage in the preparation of infant 205
173 served that the increase in solubility in the fermented foods. Fermentation has been reported as a useful and 206
REC

174 samples beyond pH 6 was not as high as in the unfer- traditional method for the preparation of low bulk 207
175 mented sample. This may be due to the exposure of weaning foods (Desikachar, 1980). 208
176 some hydrophobic groups in fermented samples, which
177 may cause reduction in solubility. The magnitude of 3.4. Water- and oil-binding capacity 209

Fermentation significantly decreased (P 6 0.05) the 210

water-binding capacity of sorghum flour by about 211
OR

Table 1
Gelation properties of fermented sorghum flour
Flour concentration % (w/v) Fermentation time (h)
0 8 16 24
UNC

2 – – – –
4 – – ± ±
6 – ± + +
8 ± + + +
10 ± + + +
12 ± + + +
14 ± + + +
16 ± + + +
18 + + + +
30 + + + +
Fig. 1. Protein solubility profiles of fermented and unfermented
sorghum flour. –, Not gelled; ±, gelled slightly; +, gelled.
 FOCH 4202 No. of Pages 5, DTD = 5.0.1
ARTICLE IN PRESS
29 July 2004; Disk Used

4 Abd Elmoneim O. Elkhalifa et al. / Food Chemistry xxx (2004) xxx–xxx

Table 2
Selected functional properties of fermented sorghum flour
Properties Fermentation time (h)
0 8 16 24
a b b
Bulk density (g/ml) 0.73 ± 0.004 0.68 ± 0.001 0.67 ± 0.000 0.66 ± 0.001b
Water-binding capacity (g/2g sample) 4.69 ± 0.064a 4.41 ± 0.071b 4.37 ± 0.049b 4.40 ± 0.000b
Oil-binding capacity (g/2g sample) 3.43 ± 0.042c 3.70 ± 0.007a 3.49 ± 0.007c 3.59 ± 0.028b
Emulsifying capacity (%) 49.39 ± 0.863b 50.79 ± 1.117ab 52.83 ± 0.467a 51.62 ± 0.474a
Emulsifying stability (%) 47.28 ± 0.403c 50.00 ± 0.000b 52.11 ± 0.735a 50.33 ± 0.460b
Foaming capacity (%) 0 0 0 0

OF
Values are means ± SD.
Values with the same superscript letter in a row are not significantly different (P 6 0.05).

212 7% after 16 h of fermentation (Table 2). In contrast it 4. Conclusion 249

PRO
213 increased the oil-binding capacity of the flour by the
214 same percentage after only 8 h of fermentation (Table The traditional Sudanese method of fermentation of 250
215 2). Water absorption capacity gives an indication of sorghum significantly improved the functional proper- 251
216 the amount of water available for gelatinization. Low- ties of sorghum flour. As previous studies have shown 252
217 er absorption capacity is desirable for making thinner that fermentation improved the nutritive value of sor- 253
218 gruels. Generally, sorghum flour, fermented and unfer- ghum flour, therefore, it would be possible to design 254
219 mented, has a higher water- and oil-binding capacity some new foods based on sorghum flour for people suf- 255
220 than flours such as raw fluted pumpkin with 0.37 g/ fering from gluten-intolerance disease. Further studies 256
221 g water-binding capacity (Giami & Bekebain, 1992) are needed in this area. 257
222 and Deturium microcarpum seed flour with 0.75 g/g
223 oil-binding capacity (Akpata & Miachi, 2001). The
TED
224 higher oil-binding capacity of sorghum flour suggests 5. Uncited reference 258
225 that this flour would be useful in formulation of foods
226 where an oil holding property is an important consid- El Nour, Peruffo, and Curioni (1998). 259
227 eration.

Acknowledgements 260
228 3.5. Emulsifying capacity and stability
REC

This work was supported by the Alexander von 261

229 The efficiency of emulsification by flour varies with
Humboldt Foundation through a Fellowship to A.O. 262
230 the type, concentration and solubility of the proteins
Elkhalifa. 263
231 (Achinewhu, 1983). Results showed that sorghum flour
232 had high emulsion activity and stability; fermentation
233 had significantly increased (P 6 0.05) the emulsion ac-
References 264
234 tivity and stability of the sorghum flour (Table 2). The
235 increase of the emulsion activity was 3%, 7% and 4% Abdel Gadir, A. M., & Mohamed, M., 1977. Kisra: Sudanese lactic/ 265
OR

236 for the three fermentation times, 8, 16 and 24 h, respec- acetic acid fermented sorghum bread. In Symposium on indigenous 266
237 tively, while the emulsion stability was increased due to fermented foods, Bankok, Thailand. 267
238 fermentation by 5% ,9% and 6% for the three fermenta- Achinewhu, S. C. (1983). Protein and food potential of African oil 268
239 tion times, respectively (Table 2). Emulsion properties bean (Pentaclethra macrophylla) and velvet bean (Mucunauries). 269
Journal of Food Science, 47, 1736–1742. 270
240 are useful in food systems such as mayonnaise, salad Akpata, M. I., & Miachi, O. E. (2001). Proximate composition and 271
241 dressing and frozen desserts. selected functional properties of Detarium microcarpum. Plant 272
UNC

Foods for Human Nutrition, 56, 297–302. 273

242 3.6. Foaming capacity Akubor, P. I., & Chukwu, J. K. (1999). Proximate composition and 274
selected functional properties of fermented and unfermented 275
African oil bean (Pentaclethra macrophylla) seed flour. Plant Foods 276
243 Sorghum flour, either fermented or unfermented, for Human Nutrition, 54, 227–238. 277
244 showed no foaming capacity at all. This was also report- Axtell, J. D., Kirleis, A. W., Hassen, M. M., Mason, N. D., Mertz, E. 278
245 ed for cowpea powder (Okaka & Potter, 1979). These re- T., & Munck, L. (1981). Digestibility of sorghum proteins. 279
246 sults showed that sorghum flour proteins in solution Proceedings of National Academy of Sciences, 78, 1333–1335. 280
247 increase the surface tension of the water and conse- Beuchat, L. R. (1976). Fungal fermentation of peanut cake. Econom- 281
ical Botany, 30, 227–234. 282
248 quently reduce the formation of foam.
 FOCH 4202 No. of Pages 5, DTD = 5.0.1
ARTICLE IN PRESS
29 July 2004; Disk Used

Abd Elmoneim O. Elkhalifa et al. / Food Chemistry xxx (2004) xxx–xxx 5

283 Coffman, G. W., & Garcia, V. V. (1977). Functional properties and dough for Kisra production. Applied Microbiology and Biotechnol- 316
284 amino acid content of a protein isolate from mung-bean flour. ogy, 37, 728–731. 317
285 Journal of Food Technology, 12, 473–484. Inyang, U. E., & Nwadimkpa, C. U. (1992). Functional properties of 318
286 Desikachar, H. S. R. (1980). Development of weaning food with high lupin seeds protein and protein concentrates. Journal of Food 319
287 calories density and low hot paste viscosity using traditional Science, 47, 491–492. 320
288 technologies. Food Nutrition Bulletin, 2, 21–23. Kinsella, J. E. (1979). Functional properties of soy proteins. Journal of 321
289 Duncan, B. D. (1955). Multiple range and multiple F tests. Biometrics, American Oil Chemistry Society, 56, 254–262. 322
290 11, 1–42. Lowry, O. H., Rosenbrough, N. J., Fair, A. L., & Randall, R. J. 323
291 Eggum, B. O., Monowar, L., Bach Knudsen, K. F., Munck, L., & (1951). Protein measurement with the Folin–phenol reagent. 324
292 Axtell, J. D. (1983). Nutritional quality of sorghum and sorghum Journal of Biological Chemistry, 193, 265–275. 325
293 foods from Sudan. Journal of Cereal Science, 1, 127–137. Mohammed, S. I., Steenson, L. R., & Kirleis, A. W. (1991). 326
294 327

OF
Elkhalifa, A. O., & El Tinay, A. H. (1994). Effect of fermentation on Isolation and characterisation of micro-organisms associated
295 protein ractions and tannin content of low and high tannin with the traditional sorghum fermentation for production of 328
296 cultivars of sorghum. Food Chemistry, 49, 265–269. Sudanese Kisra. Applied and Environmental Microbiology, 57, 329
297 Elkhalifa, A. O., & El Tinay, A. H. (1995). Effect of fermentation and 2529–2533. 330
298 germination on the in vitro protein digestibility of low and high Narayana, K., & Narsinga Rao, M. S. (1982). Functional properties of 331
299 tannin cultivars of sorghum. Food Chemistry, 54, 147–150. raw and heat processed winged bean (Psophocarpus tetragonolobus) 332
300 El Nour, I. N. A., Peruffo, A. D. B., & Curioni, A. (1998). flour. Journal of Food Science, 47, 534–538. 333

PRO
301 Characterisation of sorghum kafirins in relation to their crosslink- Okaka, J. C., & Potter, N. N. (1979). Physico-chemical and functional 334
302 ing behaviour. Journal of Cereal Science, 28, 197–207. properties of cowpea flours. Journal of Food Science, 44, 335
303 El Tinay, A. H., Abdel Gadir, A. M., & El Hidai, M. (1979). Sorghum 1235–1237. 336
304 fermented Kisra bread: Nutritive value of Kisra. Journal of the Sathe, S. K., Salunke, D. K., & Deshande, S. S. (1982). Functional 337
305 Science of Food and Agriculture, 30, 859–865. properties of lupin seeds protein and protein concentrates. Journal 338
306 El Tinay, A. H., El Mehdi, Z. M., & El Soubki, A. (1985). of Food Science, 47, 491–492. 339
307 Supplementation of fermented sorghum Kisra bread with legume Snedecor, G. W., & Cochran, W. G. (1987). In Statistical methods (7th 340
308 protein isolates. Journal of Agricutural and Food Chemistry, 21, ed.). Ames, IA: The Iowa State University Press. 341
309 679–687. Wang, J. C., & Kinsella, J. E. (1976). Functional properties of novel 342
310 Giami, S. Y., & Bekebain, D. A. (1992). Proximate composition and proteins: Alfalfa leaf protein. Journal of Food Science, 41, 343
311 functional properties of raw and processed full-fat fluted pumpkin 286–292. 344
312 (Telfariria occudentalis) seed flour. Journal of the Science of Food Yasumatsu, K., Sawada, K., Moritaka, S., Misaki, M., Tado, J., 345
TED
313 and Agriculture, 59, 321–325. Woda, T., et al. (1972). Whipping and emulsifying properties of 346
314 Hamad, S. H., Bocker, G., Vogel, R. F., & Hammes, W. P. (1992). sorghum products. Journal of Agriculture and Biological Chemistry, 347
315 Microbiological and chemical analysis of fermented sorghum 36, 719–725. 348
349
REC
OR
UNC