Effect of Maturity Stage on Protein Fractionation, In Vitro Protein Digestibility and Anti-nutrition Factors in Pineapple (Ananas comosis) Fruit Grown in Southern Sudan
Murwan K. SabahelKhier and Saifeldin A. Hussain
Department of Biochemistry, School of Biotechnology, Faculty of Science and Technology, Al Neelain University, Sudan.
E-mail: murwansabahelkhier@yahoo.com
Abstract: This investigation showed that albumin, globulin and protein content increases with increasing the days of maturity stage while glotulin and non protein nitrogen content decreases with increasing the days of maturity stage. The prolamin remained constant during the days of maturity stage. In-vitro protein digestibility improved with decreased of tannin and phytic acid content due to increasing the days of maturity stage because tannin and phytic acid inhibit the activity of pepsin enzymes.
Keywords: Pineapple, Digestibility, Ant-nutritional, Maturity and Vitamins
1.0 Introduction
The importance of pineapple fruit lies in their nutritive value. Fruit of pineapple reached the maturity stage at 105 days after flowering (Guerra and Livera, 1999).
It is rich in protein, fat, fiber, vitamins, mineral salts and water (Atif and Hagag, 1995). The net protein utilization is referred to the percentage of ingestion and utilizing protein for growth and maintenance (Osborne, et al., 1978).It is considered as supplement to daily food (Pandey and Ajanta, 1993). The physical, chemical and sensorial characters of pineapple showed significant difference at the several of maturity stage (Guerra and Livera, 1999). Pineapple contained 25-50 mg/g ellagic acid (Amakura et al., 2000).The juice of pineapple fruit contained five proteolytic enzymes collectively known as bromelain (William, 2000). Bromelain is natural blood thinner because it prevents excessive blood platelet stickiness. It also endues the thickness of mucus, which may benefit the patient with asthma or chronic bronchitis. In addition, bromelain in combination with trypsin may enhance the effect of antibiotic in people with urinary tract infection (Manhart et al., 2002). Bromelain has some remarkable characters such as its ability to reduce inflammation pain and swelling, speed the heeling of injuries trauma, surgery, sinusitis or arthritis (Mackay and Miller, 2003). The pineapple fruit contained
tannin, phenolic acid and Flavonoids (William, 1991).
Objectives of this investigation are study effect of maturity stage (75, 90 and 105
days after flowering) on the protein fractionation, in vitro protein digestibility and antinutrition factors (Tannin, phytic acid)of pineapple fruit..
2.0 Material and methods
2.1 Collection and preparation of sample: The samples were collected from Southern Sudan (Yambio Research Station, Agriculture Research Organization, Ministry of Agriculture, Sudan) basis on maturity stage. The preparation of samples was carried according to the method described by AOAC (1984).
2.2 Protein fractionation
The sequential extraction of protein was carried out according to Mendel and Osborne method (1924). It is basis on solubility of protein in different solvents. Water soluble protein (Albumins), salt soluble protein (Globulins), alcohol soluble protein (Glotulin), alkali soluble protein (Prolamin) and residual proteins (None – protein nitrogen).The residues remaining after those successive extractions with four solvents were determined by semi micro-Kjeldhal method according to AOCA (1990).Percent protein extracted was calculated to total amount of protein in the samples extracted such as follows:
Soluble protein (total) = T X N X TV X 14 X 6.25 X 100
1000 X A
Protein solubility = Soluble protein X 100
Total protein
Where:
T = Titer reading (ml of HCl), N = Normality of the HCl (0.02N), TV =Total Volume of the aliquot extracted (100ml), A = Number of (ml) of sample extracted (2.0g), 14 = each ml of HCl is equivalent to 14 mg. Nitrogen, 1000 = Number of mg in one gram and 6.25 = conversion factor from nitrogen into protein .
2.3 Anti-nutrition factors
2.3.1 Tannin content
Quantitative estimation of tannins for each sample was carried out by using modified vanillin- HCl methanol method as described by Price and Butler (1987).
There is no useful standards curve for tannin in food, but the tanninic acid was used for preparation the standard curve of tannic acid. The standard curve of tannic acid was prepared according to AOAC (1990) for measurement the concentration of tannin in our samples (plotting the concentration of tanninic acid (mg) against the corresponding reading of Spectrophotometer in Absorbance).
2.3.2 Phytic acid content
The phytic acid content was determined according to the method described by Wheeler and Ferrel (1971). Preparation of standard curve for phytic acid was done as follows: standard curve of different Fe (NO3) 3 concentrations was plotted against the corresponding of Spectrophotometer to calculated the ferric iron conc. The phytate phosphorus was calculated from the concentration of ferric iron assuming 4:6 irons: phosphorus molar ratio.
2.4 In vitro protein digestibility: It was carried out according to Maliwal method (1981) in the manner described by Monjula and John (1991) with minor modification. A know weight of the sample containing 16 mg nitrogen was taken in the triplicates and digested with one mg pepsin in 15 ml of 0.1 M HCL at 37 oC for two hours. The reaction was stopped by the addition of 15 ml (10) trichoroacetic acid. The mixture was filtrated quantatively through filter paper (Whatman No.1). Trichoroacetic acid soluble fraction was assayed for the nitrogen by semi-micro Kjeldhal method.
Protein digestibility = N2 in supernat – N2 in pepsin X 100
N2 in sample
2.5 Statistical analysis:
Three separate sub samples from each origin sample were taken and analyzed. Then mean values were averaged. Data were assessed by analysis of variance (ANOVA) as described by Gomez and Gomez (1984).
3.0 Results and Discussion
3.1 Protein fractionation
Table I indicated that albumin content of 105, 90 and 75 days maturity of pineapple is 4.4, 3.2 and 3.0 , respectively. The results revealed that albumin content increases with increasing period of maturity. Globulin content of 105, 90 and 75 days maturity of pineapple is 1.2 and 0.6, 0.4 , respectively. The finding indicated that globulin content of fruit increases with increasing the period of maturity. The Prolamin content of 105, 90 and 75 days maturity of pineapple is remained constant (0.7 ) during the period of maturity .Those results revealed that the Prolamin of pineapple not effected by period of maturity. The Glotulin content of 105, 90 and 75 days maturity of fruit is 0.9, 1.0 and 1.8, respectively. The findings revealed that Glotulin content decreases with increased the period of maturity. Non Protein N2 of 105, 90 and 75 days maturity of fruit is 92.9, 94.6 and 94.2 , respectively. All the above results are supported by the findings that reported by Amukura et al, (2000).
3.2 In vitro protein digestibility and Anti-nutrition factors
Table 2 illustrated that protein content of 105, 90 and 75 days maturity is 4.11, 3.75 and 3.70 , respectively. The findings are high than those given by Ahmed (2001).
- The protein digestibility of 105, 90 and 75 days maturity is 39.4, 36.2 and 32.2 , respectively. Those results are lower than protein digestibility of sorghum (46 ), and rice (66), maize (73 ) and wheat (81 ) that given by MacLean (1981). This finding indicated that protein digestibility of pineapple fruit is extremely poor compared with sorghum, rice, maize and wheat. Tannin content of 105, 90 and 75 days maturity is 4.5, 22.2 and 20.2 , respectively. The results revealed that high tannin content reduced the digestibility of the protein because tannin acts as anti-enzymatic activity. In addition, tannin reacts with protein to form insoluble complex compound. The phytic acid of 105, 90 and 75 days maturity is 0.40, 0.20 and 0.15 mg/g, respectively. Phytic acid interacts with protein forming complex compound and reduce the bioavailability of protein and inhibits the action of pepsin, trypsin and ? - amylase. These results explain the poor of protein digestibility in the pineapple fruit.
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Table 1: Protein fractionation of 105, 90 and 75 days maturity of pineapple fruit, grown in Southern Sudan, 2007.
Sample
Albumin
Globulin
Prolamin
Glotulin
Non Protein N2
105 days Maturity
4.4b
(±0.03)
1.2a
(±0.6)
0.7a
(±0.13)
0.9a
(±0.03)
92.9b
(±0.01)
90 days Maturity
3.2a
(±0.09)
0.6a
(±0.05)
0.7a
(±0.13)
1.0a
(±0.12)
94.6a
(±0.01)
75 days Maturity
3.0a
(±0.09)
0.4a
(±0.05)
0.7a
(±0.13)
1.8a
(±0.12)
94.2a
(±0.01)
- Mean values of same letters within the column are significantly difference at
Table 2: In Vitro protein digestibility and Anti-nutrition factors of 105, 90 and 75 days maturity of pineapple fruit, grown in Southern Sudan, 2007.
Sample
Protein
()
Protein digestibility
()
Tannin ()
Phytic acid mg/g
105 days Maturity
4.1a
(±0.01)
39.4a
(±1.1)
4.5b
(±0.07)
0.40b
(±0.02)
90 days Maturity
3.8a
(±0.05)
36.2a
(±1.2)
22.2a
(±0.13)
0.20a
(±0.01)
75 days Maturity
3.7a
(±0.05)
32.2a
(±1.2)
20.2a
(±0.13)
0.15a
(±0.01)
- Mean values of same letters within the column are significantly difference at
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About the Author
Murwan K. SabahelKhier and Saifeldin A. Hussain Department of Biochemistry, School of Biotechnology, Faculty of Science and Technology, Al Neelain University, Sudan. E-mail: murwansabahelkhier@yahoo.com
