| Food sciences |
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| Optimization of ultrafiltration separation and quality analysis of the major royal jelly proteins |
| Li Meilu1, Tan Liangliang1, Qiu Wei2, Yin Zhihong2, Shen Lirong1* |
| (1. Fuli Institute of Food Science, College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China; 2. Hangzhou Biyutian Health Food Co., Ltd, Hangzhou 311500, China) |
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Abstract Royal jelly (RJ) is the exclusive food for the Western honeybee (Apis mellifera) queen throughout its lifespan and the primary food for the honeybee larvae during the first three days of life, which is secreted by hypopharyngeal and mandibular glands of nurse honey bees. The chemical composition of RJ is very complicate, while the protein components have been found to play critical roles in RJ. Around 50% of dry RJ is made up of proteins which could be divided into two parts, water-soluble proteins and water-insoluble proteins. The water-soluble proteins, accounting for 82% to 90% of the total RJ proteins, are named major royal jelly proteins (MRJPs) which belong to the same family composed of nine members. It has been proved that MRJPs have effects such as anti-aging, immunoregulation and anti-fatigue, et al. In addition, MRJPs, especially MRJP1 are authenticated as the critical constituents to reveal the freshness of RJ and will be hopeful as one new functional component in future. In previous studies, various methods were developed for isolation of MRJPs, which included centrifugation, alkali extraction acid precipitation and column chromatography. However, a series of obstacles, such as low purity, solvent contamination and low productivity limited the application of those methods in honeybee industry.
Ultrafiltration as a novel bio-separation technology has many advantages, such as simple operation, easy scale-up, and hence has a huge potentiality for large-scale isolation of bio-macromolecules. Our effort was to obtain the separation technology of MRJPs by using ultrafiltration and provide new knowledge for the industrial application of RJ process in future. Since the molecular masses of MRJPs are in the range of 4.9×1048.7×104, the poly-sulfone hollow fiber membrane component used in the ultrafiltration equipment was comprised of two types with different molecular mass cut-off membranes. The ultrafiltration membrane with a 1×105 molecular mass cut-off was employed to remove macromolecular impurities, whereas the 4.9×104 cut-off membrane was employed to retain most of MRJPs. In this study, disposals were taken to separate soluble proteins, MRJPs from RJ by ultrafiltration. The control process included pH, ion intensity and ratio of water to material. In order to obtain the optimal conditions to extract MRJPs, orthogonal test was employed during investigation on experimental factors. The results showed that the optimal condition was as follows, ratio of water to material 5∶1, ion intensity 0.5 mol/L and pH 7.0. Under the optimal condition, 82.63% of MRJPs was extracted from fresh RJ. It was determined that the lyophilized powder of MRJPs contained 91.00% soluble protein, 0.01% moisture, 2.00% 10-HDA and 15.22% total sugar. Besides, the by-products contained 10-HDA and total sugar were recycled, and the recycling rate of the two components reached 3.60% and 2.11%, respectively. This method with many advantages, such as no solvent pollution and fully utilization of 10-HDA, has provided a new technology for MRJPs separation by first using ultrafiltration in deep process of RJ, and will be hopeful for large scale application in RJ process industry.
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Published: 20 September 2014
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蜂王浆主蛋白(MRJPs)超滤分离工艺优化及产物质量分析
采用超滤方法从鲜王浆中提取王浆主蛋白（major royal jelly proteins,MRJPs）,先就水料比、pH、离子强度3种单因素对MRJPs提取率的影响进行单因素分析,然后通过正交试验对MRJPs的超滤提取工艺进行优化,最后对MRJPs分离产物冻干粉和滤后液进行成分测定。结果表明:最佳提取工艺为水料体积质量比5∶1 (mL/g),pH 7.0,离子强度0.5 mol/L。在最佳工艺条件下,MRJPs的提取率为82.63%,冻干粉含可溶性蛋白91.00%、水分0.01%、总糖15.22%、王浆酸2.00%,超滤副产物——滤后液中王浆酸和总糖的回收率分别为3.60%和2.11%。超滤工艺具有无溶剂污染、王浆成分可回收利用,适合规模化生产的优点,为MRJPs分离提取提供了新的工艺方法,为王浆深加工和MRJPs蛋白产品开发提供了科学依据。
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