Food Engineering
Hossein Mohebodini; Atefe Maqsoudlou
Abstract
Introduction: Microencapsulation is the most commonly used method of preserving proteins and peptides, which increases the stability in different conditions. Bee pollen with 10–40% protein, is a valuable source of protein that has functional and nutraceutical properties. By hydrolysis and producing ...
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Introduction: Microencapsulation is the most commonly used method of preserving proteins and peptides, which increases the stability in different conditions. Bee pollen with 10–40% protein, is a valuable source of protein that has functional and nutraceutical properties. By hydrolysis and producing bioactive peptides, their functional and health effects will be improved. Fibersol is a dietary fiber that can be used in many foods and supplements. This carbohydrate compound is actually non-digestible maltodextrin and has recently been used as a wall material in encapsulation. Few studies have conducted on the microencapsulation of hydrolysed proteins and their stability during accelerated conditions. On the other hand, by-products of honey bees such as pollen have been less noticed; therefore the aim of this study was the microencapsulation of bioactive bee pollen protein hydrolysate by fibersol and WPC and to study the changes of structure and stability of resulted microcapsules during the exposure to UV radiation. Material and Methods: Bee pollen was hydrolysed by Alcalase (1.5%) for 4 h in shaking incubator. The protein hydrolysate was microencapsulated using WPC, fibersol, and their combination by spray drying. The wall materials and hydrolysed protein were used in ratio of 10:1 (w/w). WPC 2%, fibersol 2%, as well as WPC and fibersol mixtures with 1:3 ratio, were the wall materials. For accelerating the oxidation reactions, the obtained capsules were exposed to UV radiation for 48 h. During the exposure to UV radiation, the DPPH radical scavenging activity of microcapsules and hydrolysed protein was measured. Interactions between hydrolysed protein compounds, WPC and fibersol were identified by the FTIR spectroscopy. The SEM was used to investigate the morphology of the microcapsules. Results & Discussions: Almost at all experimental time, the highest DPPH radical scavenging during exposure to UV radiation was related to the capsules prepared using fibersol and WPC mixture and after that the capsule with WPC as wall material. The FTIR spectroscopy of the hydrolysed protein was changed significantly when it was exposed to UV radiation. This change caused by losing the hydrogen bonds in the secondary structure of proteins, including the separation of two polypeptide chains or the opening of the αhelix and loss of β-sheet structure. The FTIR profile of capsulated hydrolysed protein by fibersol showed that the adhesion of protein and polysaccharide changed the absorbance of C–H bending and N–H stretching bands of amide groups in the hydrolysed protein in 3000–3500 cm-1 and the stretching band of C–H and O–H group in the region of 2000–3000 cm-1 for fibersol in the wall. After exposure to UV, because of cross-linking in fibersol and more involving the molecules of fibersol to protein, the absorbance was increased in the region of 1500–3500 cm-1. The number of peaks and absorbance in the FTIR spectra of hydrolysed proteins microencapsulated in WPC were more than number of peaks and absorbance in the FTIR spectra of WPC. There was no significant difference in the FTIR spectra of hydrolysed protein encapsulated with WPC before and after exposure to UV. The peaks in FTIR spectra of hydrolysed protein microencapsulated with the mixture of WPC and fibersol, showed higher absorbance level than the peaks of fibersol and lower than peaks of WPC. None of the peaks of microencapsuls with the wall of mixture of WPC and fibersol, were changed after exposure to UV radiation. Results of SEM showed that the microcapsules prepared with mix of fibersol and WPC had a uniform and smoother wall than microcapsules prepared with only fibersol. Finally, the mix of WPC and fibersol was selected as the best wall with a proper protective ability for the microencapsulation of hydrolysed proteins and protection against UV radiation.
Atefe Maqsoudlou; Alireza Sadeghi Mahoonak; Mohammad Ghorbani; Fidel Toldta
Abstract
Introduction: Bee pollen, commonly referred as the ‘‘life-giving dust’’, results from the agglutination of flower pollens with nectar using salivary substances of the honeybees (Almeida-Muradian et al., 2005). Pollen contains 10 to 40% protein, 1 to 13% lipid, 13 to 55% carbohydrates and 2 to ...
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Introduction: Bee pollen, commonly referred as the ‘‘life-giving dust’’, results from the agglutination of flower pollens with nectar using salivary substances of the honeybees (Almeida-Muradian et al., 2005). Pollen contains 10 to 40% protein, 1 to 13% lipid, 13 to 55% carbohydrates and 2 to 6% minerals. Royal Jelly is produced by enzymatic digesting of bee pollen by proteases and other natural enzymes. Based on dry weight, it contains 27-41% protein, 30% carbohydrates, 8-19% lipids, minerals, trace elements and some vitamins (Sabatini et al., 2009; Wytrychowski et al., 2013). The antioxidant properties of royal jelly and bee pollen, are related to main proteins and phenolic compounds and flavonoids (Nagai and Inue, 2004). The antioxidant activity of peptides can be evaluated using DPPH, radicals scavenging activity, Ferric reducing, Ferrous chelating activity (Khantaphant et al., 2011). Antioxidant and ACE inhibitory activity of pollen, royal jelly and peptides were investigated by different researchers (Bogdanov, 2014; Morais et al., 2011; Salampessy et al., 2015; Marinova and Tchorbanov, 2010; Wiriyaphan et al., 2012). The objective of present research was optimization of enzymatic hydrolysis of bee pollen protein by Alcalase according to its antioxidant and ACE inhibitory activity compared to royal jelly.
Materials and methods: The preparation of the bee pollen extract was performed by mixing the bee pollen with water (1:10) (w/v). The macerates were filtered and centrifuged at 12000 g. The obtained supernatant was lyophilized. The royal jelly extract were prepared using method described by Liu et al., 2008. The total phenolic content of the extracts was recorded using the Folin–Ciocalteu method (Moreira et al., 2008). DPPH radical-scavenging activity was determined as described by Bersuder, Hole, and Smith (1998). The ability of the hydrolysate to reduce iron (III) was determined according to the method of Bougatef et al. (2008). Bee pollen was added and homogenized with 5 volumes of distilled water. pH and temperature of the solution were adjusted to pH=8 and 50◦C. Alcalase in the concentration range of 1 to 2% w/w were added to the pollen protein solution. Enzymatic hydrolysis performed during different times 2 to 5 hours. Hydrolysis was stopped by heating at 80˚C for 10 min. The hydrolysats were centrifuged at 4000x g for 30 min to remove the residue. The supernatants were pooled and then lyophilized (Matsuoka et al., 2012). DPPH radical scavenging ability and reducing power of pollen hydrolysates of pollen hydrolysates were measured. Also ACE-inhibitory activity of pollen hydrolysates was measured was assayed by method reported by Nakamura et al. (1995). Statistical analysis of results before hydrolysis was done by SPSS. Optimization of enzymatic hydrolysis was done by Response Surface Methodology (RSM) in Design Expert software.
Results and discussion: Total phenol value measured for pollen ranged between 48.15 to 174 mg Gallic acid/g for royal jelly ranged from 9.24 to 87.261 mg Gallic acid/g. Considering that royal jelly is obtained by direct digestion of pollen, the amounts of their phenolic compounds were comparable (Bogdanov, 2014). Phenolic compounds increased by increasing concentration royal jelly and pollen extract in dose dependent manner. Increasing concentrations of royal jelly in range of 300 to 1000 mg/l was more effective than pollen (p
