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Effect of heat and alkaline hydrolysis on the amino acid profile of Jatropha curcas seed cake.


One of the major challenges to increased and sustainable livestock production in Africa and Nigeria in particular is lack of affordable protein in the animal feeds. The protein sources such as soybean, cotton, rape, sunflower and peanut seed cakes are either not readily available or costly for subsistence farmers. Fortunately Jatropha curcas, which is a drought-resistant shrub belonging to the Family Euphorbiaceae, is a good substitute as a protein source for livestock feeds. This is because besides being easy to grow, the crop seeds are oily and highly proteineous thereby making it a cheap protein source. Nevertheless, J. curcas seeds are toxic, and thus have to be detoxified prior to supplementing as a protein source in animal feeds [1-3]. Recently, J. curcas has attracted attention of various research organizations, governments, public and international developmental agencies and industries in the tropics and subtropics due to its adaptability to semi-arid marginal sites, the possibility of using its oil as a diesel fuel substitute and its role in erosion control [4]. The seeds of J. curcas are a good source of oil, yielding between 40-80 % oil [5-9]. Although the seed cake meal is rich in protein, it is toxic to rats, mice, ruminants and humans due to the presence of antinutritional factors such as phorbol esters, curcin, trypsin inhibitors, lectin, [5, 6, 8, 10-12], .thus its use as food or feed source has not been encouraging. Recent findings indicate that after proper detoxification process the seed meal can serve as a protein substitute in feed meals of animal feeds [5, 13].

Consequently, several works has been carried out on the J. curcas seed so that it can be used as a source of protein in animal feed. For example Oladele and Oshodi [8] attempted the detoxification of the seeds using local fermentation process while Marti'nez-Herrera et al. [13] also used chemical such NaHC[O.sub.3], ethanol as well as irradiation as a method of detoxification. However, Aregheore et al. [10] reported that heat and chemical (ethanol) treatments were able to reduce the antinutrient factors in J. curcas seed to a tolerable minimum while solid state fermentation employed by Belewu and Sam [14] was able to detoxify and inactivate almost 100 % of the antinutrient contents of Aspergillus niger treated sample of Jatropha kernel cake to a tolerable level [10, 14]. Similarly Usman et al. reported the effect of alkaline hydrolysis on the quantity of extractable protein fractions in Jatropha treated seed cakes [15]. In the work reported by Usman et al. [15], it was concluded that the treatment increase the quantity of prolamin in all the treated cake, while the quantity of globulin, albumin and glutelin were reduced.

Since the quality of any protein source relates to its amino acid composition, digestibility, bioavailability and ability to supply the essential amino acid in the amount required by the organism consuming it. Based on these facts there are two methods of assessing the quality of proteinous species, amino acid analysis and feed test. In view of the foregoing, the amino acid profile of heat and chemically treated Jatropha cucas seed cake in comparison to the untreated cake was investigated.


Preparation of seed cake

Jatropha curcas seeds were obtained from ripe fruits harvested from different locations in Ilorin, North-Central, Nigeria. The seeds were dehulled and milled with magnetic blender (SHB-515 Model, Sorex Company Limited, Seoul, Japan). Standard Official and Tentative Method of oil Chemists Society procedure was used to defat the seed cake [16]. The defatted seed cake was dried and kept for analysis.

Alkaline hydrolysis and chemical treatment of the seed cake

A method similar to that of Aregheore et al. [10] was adopted for the hydrolysis of the seed cake. In the method, five portions of 60 g of the seed cake were separately moistened with 10 ml 1-5 M NaOH solutions in separate beakers and left to stand for 24 hrs. Each mixture was milled into a paste using glass rod, and was subsequently covered with aluminum foil, placed in an autoclave at 121[degrees]C for 30 min. The autoclaved samples were removed and allowed to cool to room temperature. Each of the samples was then put into different white clothes, immersed in a jar of distilled water and mixed thoroughly. This was followed by squeezing and subsequent immersing in another jar containing ethanol for 30 minutes, the sample was squeezed and spread to dry at room temperature. A 60 g of the seed cake which was not treated with NaOH, water and ethanol was also kept and labeled as untreated cake.

Determination of crude protein

The crude protein (CP) of the untreated, defatted and chemically treated Jatropha meals was determined in accordance to AOAC procedure [17].

Determination of Amino Acid Profile

The amino acid composition of the treated and untreated J. curcas seed was determined using Technicon Amino Acid Analyzer (TSM-1 Technicon Instrument, Basingstoke, Hampshire, UK) using Norleucine as internal standard. This procedure follows the method described by Bassler and Buchholz [18]. The contents of different amino acids recovered were presented in g/100 g of protein. The amino acid contents of the total seed proteins were compared with the FAO/WHO reference pattern, soya bean and reported literature values for J. curcas [4, 11, 19-21].

Statistical Analysis

All data collected for the concentrates were subjected to analysis of variance, ANOVA. Means were compared using Duncan's multiple range tests at P<0.05 confidence level.


The defatted seed cake of J. curcas has been subjected to chemical and heat treatment in order to improve its nutritional quality. The percentage crude protein in the treated and untreated seed cake is presented in Table 1. From Table 1, it is observed that the chemical and heat treatment has improved the quantity of crude protein in the seed cake compared to the untreated sample. The crude protein content of the untreated sample was 63.02 % while seed cake treated with 1 M, 2 M and 3 M NaOH gave crude protein contents of 70.53, 71.46 and 67.76 %, respectively Table 1. The data presented in Table 1 also revealed that the percentage crude protein content for the seed cake treated with 4 M and 5 M NaOH were lower than that of the untreated seed cake, the reduction in the quantity of the crude protein content may be due to high concentration of NaOH leading to loss of some amino acids in the sample.

The amino acid composition of the treated and untreated seed cake is presented in Table 2. The table data presented in Table 2 revealed that the amino acid profiles of the treated seed cakes were similar and comparable to the values reported for different provenances of J. curcas, (Cape Verde and Nicaragua) [21]. The trend for the non-essential amino acids is similar to that recorded for the essential amino acids (Table 3). The values increase from 1 M to 2 M and decrease steadily from 3 M to 5 M NaOH treated seed cakes, the exception being the values for histidine and aspartic acid whose values do not follow a regular pattern. From the global perspective of the data presented, the values for the treated seed cakes were higher than those of the untreated cakes. Table 4 presents the leucine to lysine values for the treated and untreated J. curcas seed cakes, soybean and the WHO/FAO recommended standards. The leucine to lysine ratio of the untreated and treated J. curcas seed cakes are lower than the marginal value and compare favourably with the WHO/FAO standard and the soybean values.


The observation that the heat and chemical treatment does not adversely affect the crude protein contents of treated J. curcas indicates that this method may be successfully used to detoxify J. curcas seed cake. However, the highest crude protein value of 71.46 % was obtained with the cake treated with 2 M NaOH, thereby suggesting that this concentration might be a better method of treating Jatropha seed cake. This value is comparable to the one obtained by Martinez-Herrera et al [4] in which four different methods were employed for the detoxification of Jatropha seed cake from four different locations and found that the chemical and heat treatment method gave the best of 70.9 % crude protein content in the seed from Mexican city of Yautepec, Morelos state [4].

It has been reported that protein solubility in neutral salt solutions depends on ionic strength and pH of the medium used for the extraction [22]. Increase in ionic strength of any salts solution may reduce the solubility of protein fractions. The effect of pH on the solubility of protein depends on whether the desired protein is at its isoelectric pH. Some protein fractions are at their minimum solubility at isoelectric pH, while some are completely insoluble at this pH. This fact may explain the low crude protein content obtained at high concentration of NaOH.

Results present in Table 2 indicate that there was increase in the essential amino acids of the treated J. curcas seed cake with 2 M NaOH treated seed cake recording the highest values. The only exception being cystine whose value was the same for both the treated (at 1 M and 2M NaOH) and untreated seed cakes. In all, the value increases from 1 M to 2 M NaOH and then decreases steadily from 3 M to 5 M NaOH treated seed cakes. However, a steady decrease from 1 M to 5 M NaOH treated cake was recorded for cystine.

Generally, levels of essential amino acids for the J. curcas treated seed cakes were higher than the WHO/FAO recommended standards with the exception of cystine and methionine in which lower values were obtained (Table 2). In comparison with the amino acid profile of soybean, it is observed that the values obtained for the J. curcas treated seed cake were higher than those for soybeans. The exceptions being the values for isoleucine, tyrosine and cystine in which the data reported for soybeans were higher than those obtained for the J. curcas treated seed cake (Table 2). This observation indicates that the nutrition value of J. curcas treated cake compare favourably with that of soybeans. This observation has already been reported by Makkar and Becker [11].

It has been established that utilization of lysine and isoleucine in protein is affected by the amount of leucine present [20]. A leucine to lysine ratio greater than 4.6 will hinder the utilization of lysine. Results present in Figure 1 indicate that the value is not only lower than 4.6 but it is also lower than those of some Nigerian grown cereals [23]. Hence the lower ratio will no doubt make the treated J. curcas seed cake to be nutritionally superior to commonly grown Nigerian cereals.



The world has shifted sourcing protein for animal diets and oils for industrial raw material to oil seeds. Notable among the oil seeds is Jatropha curcas, from the researches carried out so far on J. curcas, the seed has nutritional potential that compares favorably to conventional oil seeds and protein source (soybean). The levels of essential amino acids except cystine and methionine were comparable with that for the FAO reference protein. The seed can thus be used as an alternative protein source in animal feed formulation after it has been properly detoxified. If well processed, it would reduce competition between man and livestock for the conventional sources of proteins. Thus, cultivation of this potentially rich plant is encouraged in order to reduce over-dependence on the currently limited sources of protein and oil.


[1.] Aderibigbe AO, Johnson COLE, Makkar HPS, Becker K and N Foidl Chemical composition and effect of heat on organic matter- and nitrogen-degradability and some antinutritional components of Jatropha meal. Animal Feed Science Technology, 1997; 67: 223-243.

[2.] Ahmed OMM and SEI Adam Toxicity of Jatropha curcas in sheep and goats. Res. Vet. Sci., 1979; 27: 89-96.

[3.] Becker K and HPS Makkar Toxic effects of phorbolester in carp (Cyprinus carpio L.), Vet. Hum Toxicol. 1988; 40: 82-86.

[4.] Martinez-Herrera J, Martinez Ayala AL, Makkar H, Francis G and K Becker Agroclimatic Conditions, Chemical and Nutritional Characterization of Different Provenances of Jatropha curcas L. from Mexico. Eur. J. Sci. Res, 2010; 39(3): 396-407.

[5.] Abou-Arab AA and FM Abu-Salem Nutritional quality of Jatropha curcas seeds and effect of some physical and chemical treatments on their anti-nutritional factors. Afr. J. Food Sci. 2010; 4(3): 93-103.

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[7.] Makkar HPS, Becker K, Sporer F and M Wink Studies on Nutritive Potential and Toxic Constituents of Different Provenances of Jatropha curcas. J. Agric. Food Chem. 1997; 45: 3152-3157.

[8.] Oladele EOP and AA Oshodi Effect of fermentation on some chemical and nutritive properties of Berlandier Nettle spurge (Jatropha cathartica) and physic nut (Jatropha curcas) seeds. Pak. J. Nutr. 2008; 7: 292-296.

[9.] Waraporn A, Pilanee V, Phanu S and M Taweesiri Optimization of Protein Hydrolysate Production Process from Jatropha curcas Cake. World Acad. Sci. Eng. Tech. 2009; 53: 109-112.

[10.] Aregheore EM, Becker K and HPS Makkar Detoxification of a toxic variety of Jatropha curcas using heat and chemical treatments, and preliminary nutritional evaluation with rats. S. Pac. J. Nat. Sci. 2003; 21: 50-56.

[11.] Makkar HPS and K Becker Potential of J. curcas seed meal as a protein Supplement to Livestock Feed. Constraints to its Utilization and Possible Strategy to Overcome Constraints. In: Gubitz GM, Mittelbach M, Trabi M (Eds). Biofuel and Industrial Products from Jatropha curcas, Proceedings of the Symposium "Jatropha 97" held in Managua, Nicaragua, Feb 23-27; 1997; 190-205.

[12.] Makkar HPS, Francis G and K Becker Protein concentrate from Jatropha curcas screw-pressed seed cake and toxic and antinutritional factors in protein concentrate. J. Sci. Food Agric. 2008; 88:1542-1548.

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[15.] Usman LA, Ameen OM, Lawal A and GV Awolola Effect of Alkaline Hydrolysis on The Quantity of Extractable Protein Fractions (Prolamin, Albumin, Globulin and Glutelin) in Jatropha Curcas Seed Cake. J. Food Tech. 2008; 6(6): 259-262.

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[22.] Charles A and L Guy Food Biochemistry, Aspen Publishers Inc., Gaithersburg, Maryland. 1999, pp. 71-89.

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Ameen OM * (1); Usman LA (2); Muhammed NO (1); Okeola OF (1); Boluwarin EO (1) and OO Fadeyi (1)

* Corresponding author's email:;

(1) Chemistry Department, University of Ilorin, Ilorin, Nigeria

(2) Biochemistry Department, University of Ilorin, Ilorin, Nigeria
Table 1: Percentage crude protein in the treated and untreated
Jatropha curcas seed cake

Sample                % Crude Protein

Untreated seed cake   63.02 [+ or -] 0.07

caketreated   1 M     70.53 [+ or -] 0.06
              2 M     71.46 [+ or -] 0.06
              3 M     67.76 [+ or -] 0.08
Treated       4 M     60.82 [+ or -] 0.07
samples       5 M     56.19 [+ or -] 0.05

Table 2: Essential amino acid compositions of treated and untreated
of J. curcas seed cake, soybean and essential amino acid pattern
suggested by FAO/WHO (g/100 g protein)


Amino acid     Untreated              Treated seed cake

               seed cake   1 M     2 M     3 M     4 M     5 M

Isoleucine     3.74        4.12    4.69    3.30    3.23    3.04
Leucine        7.54        6.12    8.46    7.59    5.77    4.26
Lysine         6.86        7.35    7.62    6.27    5.08    4.86
Phenyalanine   5.14        5.66    5.66    4.71    4.62    3.08
Tyrosine       2.86        2.86    3.17    2.54    2.22    1.90
Cystine        0.66        0.66    0.66    0.53    0.40    0.26
Methionine     1.30        1.09    1.30    0.89    0.83    0.63
Threonine      4.06        4.58    4.36    4.02    3.86    3.37
Tryptophan     ND          ND      ND      ND      ND      ND
Valine         5.60        5.27    6.24    5.27    4.33    3.05

Amino acid     WHO/FAO     Soybean


Isoleucine     4.2         4.8
Leucine        4.2         8.0
Lysine         4.2         6.4
Phenyalanine   2.8         4.8
Tyrosine       2.8         3.2
Cystine        2.0         0.8
Methionine     2.2         0.9
Threonine      2.8         4.0
Tryptophan     1.4         1.3
Valine         4.2         4.8

ND = Not Determined

Table 3: Non-essential amino acid compositions of treated
and untreated of J. curcas seed cake

Amino acid                              Samples

                Untreated               Treated seed cake

                seed cake   1 M      2 M     3 M      4 M      5 M

Histidine       2.14        2.27     1.89    2.25     1.64     1.20
Arginine        5.87        4.66     6.73    4.31     4.14     3.62
Aspartic acid   9.74        10.05    9.80    10.20    10.17    9.99
Serine          3.80        4.07     4.61    3.80     3.36     3.15
Glutamic acid   13.63       13.93    14.8    14.69    11.51    10.15
Proline         3.26        3.05     3.66    3.15     3.46     2.44
Glycine         3.65        3.56     3.70    3.70     3.07     1.68
Alanine         3.49        3.65     4.18    3.42     3.04     2.35
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Article Details
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Title Annotation:Short Communication
Author:Ameen, O. Mubarak; Usman, L.A.; Muhammed, N.O.; Okeola, O.F.; Boluwarin, E.O.; Fadeyi, O.O.
Publication:African Journal of Food, Agriculture, Nutrition and Development
Article Type:Report
Geographic Code:6NIGR
Date:Apr 1, 2014
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