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Ultrafiltration membrane separation method for obtaining royal jelly major protein and active filtrate in royal jelly

The technology of a royal jelly main protein and a separation method is applied in the field of separating the royal jelly main protein and active filtrate from royal jelly, which can solve the problems of no advantages and difficult optimization conditions.

Active Publication Date: 2014-05-07
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of ultrafiltration membrane separation technology for royal jelly needs to involve many factors, and it is difficult to obtain optimal conditions. Compared with chromatography, centrifugation, dialysis separation and purification technology, there is no public report (the literature generally only discloses success) case)

Method used

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  • Ultrafiltration membrane separation method for obtaining royal jelly major protein and active filtrate in royal jelly
  • Ultrafiltration membrane separation method for obtaining royal jelly major protein and active filtrate in royal jelly
  • Ultrafiltration membrane separation method for obtaining royal jelly major protein and active filtrate in royal jelly

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] MRJPs ultrafiltration separation and concentration in fresh royal jelly in embodiment 1

[0054] 1. Using ultrafiltration membrane to separate and concentrate MRJPs from fresh royal jelly

[0055] Principle: Since royal jelly usually contains 11%-14.5% royal jelly crude protein and 20%-30% sugar, the molecular weight of royal jelly main protein is 49-87kDa, and the molecular weight of sugar is 150-200. The advanced ultrafiltration membrane can efficiently intercept royal jelly main protein, and can effectively separate it from sugar and other low molecular weight components to achieve the purpose of purification and concentration.

[0056] Operation process: use figure 1 The ultrafiltration device shown (feed liquid tank 1, circulating pump 2, membrane module 3, and pressure regulating valve 4 are connected in sequence), press figure 2 Method flow, operation. Weigh 50g of fresh royal jelly, mix according to a certain proportion of material and water, fully stir at 4...

Embodiment 2

[0082] Example 2 Detection of Ultrafiltration Separation of MRJPs Products

[0083] 1. SDS-PAGE detection of MRJPs products separated by ultrafiltration

[0084] Fresh royal jelly was obtained and separated by ultrafiltration under optimal conditions to obtain MRJPs products for SDS-PAGE electrophoresis detection. The concentration of the electrophoresis stacking gel was 5%, the concentration of the separating gel was 12%, the electrophoresis buffer system was Tris-glycine buffer, and the protein sample loading volume was 20 μL / well.

[0085] ① Preparation of separation gel: 12% separation gel (5mL): 1.6mL ddH20, 2.0mL 30% acrylamide, 1.3mL1.5mol / LTris-HCL (pH8.8), 50μL 10% SDS, 50μL 10% ammonium persulfate, 4μL TEMED;

[0086] ② Prepare stacking gel: 5% stacking gel (1mL): 0.68mL ddH20, 0.34mL 30% acrylamide, 0.26mL 1.0mol / LTris-HCL (pH6.6), 10μL 10% SDS, 10μL 10% ammonium persulfate, 4μL TEMED;

[0087] ③Sampling: Mix 2×protein buffer with the same amount of protein sample...

Embodiment 3

[0103] 10-HDA and total sugar recovery in the filtered liquid of fresh royal jelly in embodiment 3

[0104] 1. Recovery of 10-HDA and total sugar in the filtrate

[0105] Under optimal conditions, the ultrafiltration membrane system was used to process the concentrated MRJPs of fresh royal jelly to obtain filtrate samples. First use a rotary evaporator to concentrate the filtrate at 40°C (30 mL of concentrated sample can be obtained per 100 mL of filtrate), and then freeze-dry in a vacuum to obtain a lyophilized powder.

[0106] 2. Determination of recovery rate of fresh royal jelly and ultrafiltration liquid 10-HDA

[0107] Referring to the 10-HDA determination method in GB9697-2008, the 10-HDA content of the lyophilized product of the filtrate and the fresh royal jelly were measured. The results (Table 4) showed that the 10-HDA contents of the lyophilized product of the fresh royal jelly and the filtrate were respectively 1.13% and 0.93%. follow the next

[0108] Calcula...

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Abstract

The invention discloses an ultrafiltration membrane separation method for obtaining royal jelly major protein and active filtrate in royal jelly. The method comprises the following steps: weighing royal jelly, stirring and mixing uniformly with water at the temperature of 4 DEG C, regulating pH by using NaOH, regulating the ion strength by using NaCl, and removing particles by using a 0.2mu m microfiltration membrane; separating and removing macromolecular impurities by using an ultrafiltration membrane M1 with the molecular weight cutoff of 100kDa, separating and concentrating the royal jelly major protein in the M1 filtrate by using an ultrafiltration membrane M2 with the molecular weight cutoff of 49kDa, supplementing purified water when the royal jelly major protein is concentrated to be 1 / 3 volume, allowing a small molecule solution to pass through the M2, wherein the obtained cutoff solution is a purified and concentrated royal jelly major protein solution, and performing freeze drying to form the royal jelly major protein freeze-dried powder. The small molecule solution contains polysaccharides and 10-hydroxy-2-caproleic acid (10-HDA) and can be used for production of functional drinks. The royal jelly major proteins (MRJPs) are separated and concentrated by using an ultrafiltration device, the extraction rate of the MRJPs is 82.63 percent, and the royal jelly major protein is subjected to freeze-drying to form the freeze-dried powder with the protein content of more than 70 percent. Moreover, the filtrate is used for preparing drinks, and comprehensive utilization of the royal jelly is realized.

Description

technical field [0001] The invention relates to the fields of ultrafiltration and immunology, in particular, the invention relates to a method for separating royal jellybumin and active filtrate from royal jelly. Background technique [0002] Royal Jelly (Royal Jelly) is secreted by the royal jelly glands (tongue gland and jaw gland) on the head of 6-12-day-old worker bees. It is used to feed the special milky substance that feeds the queen bee and larvae. Royal jelly is rich in protein, among which the main water-soluble protein, Major Royal Jelly Proteins (MRJPs), accounts for 46% to 89% of the total protein in royal jelly. MRJPs include nine proteins, MRJP1-9, with a molecular weight of 49-87 kDa, among which MRJP1 is the most abundant, accounting for about 50%. MRJPs have physiological and nutritional functions such as promoting cell growth, anti-tumor, and antibacterial. On April 24, 2011, Dr. Masaki Kamakura of Toyama Prefectural University in Japan reported for the ...

Claims

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Application Information

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IPC IPC(8): C07K14/435C07K1/34A23L2/38
CPCA23L2/38A23L2/74A23L21/20C07K14/43572A23V2002/00A23V2250/542
Inventor 沈立荣李玫璐于张颖周志军谭量量赵敏洁尹志红裘卫
Owner ZHEJIANG UNIV
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