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Extracting and purifying method of chlorella lutein protein complex

A purification method, the technology of chlorella, applied in the biological field, can solve the few problems involving lutein protein complexes, etc., and achieve the effect of easy production scale-up and strong stability

Active Publication Date: 2014-03-05
FUZHOU UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the research on lutein in Chlorella is mainly focused on the extraction of lutein from Chlorella and its physiological effects, while there are few related studies involving lutein-protein complexes

Method used

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  • Extracting and purifying method of chlorella lutein protein complex
  • Extracting and purifying method of chlorella lutein protein complex
  • Extracting and purifying method of chlorella lutein protein complex

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Embodiment 1

[0022] Step 1: Weigh 50g of chlorella algae mud frozen at -20°C, melt it at 4°C, add a small amount of distilled water, centrifuge at 5000r / min for 10min at 4°C, and collect the precipitate. The precipitate was washed by centrifugation three times under the same conditions. Add 50mL phosphate buffer (20mM, pH 7.0) to the cleaned algae mud, mix well and perform ultrasonic crushing in an ice-water bath (ultrasonic conditions: power 600W, ultrasonic time: 30min, operation 2s, intermittent 3s). After crushing, centrifuge at 4°C and 5000r / min for 10min to remove unbroken cells, then centrifuge at 14000r / min for 30min to collect the precipitate, which is the thylakoid membrane.

[0023] Step 2: suspend thylakoid membrane precipitation with 20mM, pH7.0 phosphate buffer, adjust the content of chlorophyll a to 1.5mg / mL, add Tween-20 to make the final concentration mass fraction reach 1%, and stir slowly at 4°C After solubilizing in the dark for 12 hours, centrifuge at 12,000 r / min for...

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Abstract

The invention relates to an extracting and purifying method of a chlorella lutein protein complex. The extracting and purifying method comprises the following steps: carrying out wall-breaking treatment onto chlorella by utilizing an ultrasonic crushing method; carrying out solubilization onto chromoprotein on a thylakoid membrane by adopting a surfactant; and purifying the lutein protein complex by adopting ammonium sulfate classification and sedimentation, ion exchange chromatography and gel filtration chromatography. A pigment composition of the complex is analyzed by utilizing full-wavelength scanning and high performance liquid chromatography, wherein the obtained lutein protein complex has remarkable ABTS radical-scavenging activity. The extracting and purifying method disclosed by the invention can ensure structure and function stability of the complex by utilizing surfactant solubilizing chromoprotein; purified lutein protein complex obtained by column chromatography is simple in purifying process and convenient to produce and expand; and the lutein existing in the form of the complex not only improves stability, but also keeps antioxidant activity.

Description

technical field [0001] The invention relates to a method for extracting and purifying a chlorella lutein protein complex, belonging to the field of biotechnology. Background technique [0002] Lutein, also known as "plant lutein", is a class of naturally occurring oxygen-containing carotenoids, which are widely found in higher plants and algae. A series of significant physiological activities such as sclerosis. However, due to poor water solubility and instability to light and oxygen, lutein monomer has certain limitations in application. Due to its own hydrophobicity, naturally occurring lutein can form a lutein-protein complex by interacting with the hydrophobic domain of lipoproteins, thereby maintaining its own stability and exerting physiological activity. In higher plants and algae, light-harvesting pigments such as lutein and chlorophyll are combined with membrane proteins to form a light-harvesting pigment-protein complex, which is located on the thylakoid membrane...

Claims

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Application Information

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IPC IPC(8): C07K14/405C07K1/36C07K1/30C07K1/18C07K1/16
CPCC07K14/405
Inventor 汪少芸林祥志蔡茜茜杨倩赵立娜张林杨善军荣辉王昭凯
Owner FUZHOU UNIVERSITY
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