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Enzyme method modified preparation method for improving activity of lily polysaccharides

A technology of lily polysaccharide and activity is applied in the field of biomodification preparation for improving the activity of lily polysaccharide, which can solve the problem of low activity of lily polysaccharide, and achieve the effects of improved immune activity, strong specificity and controllable reaction.

Inactive Publication Date: 2013-12-11
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the problem of low activity of lily polysaccharides and provide a biomodification preparation method for improving the activity of lily polysaccharides

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] According to the following conditions and operations, the complex enzymes were used to modify the lily polysaccharide: substrate lily polysaccharide concentration 30 / L; β-mannanase concentration 500U / L, β-glucanase 250U / L; pH 5.5 (PBS buffer liquid); temperature 40°C; time 0.5h. After the reaction, heat the solution to 100°C, keep it warm for 5min, then centrifuge (6000rpm) to remove the enzyme protein, put the supernatant into a dialysis bag (molecular weight cut-off: 5.0kDa) and dialyze (4°C, 12h), after dialysis The sample was centrifuged (5000rpm), and the supernatant was freeze-dried to obtain a white powder lily polysaccharide modified product (molecular weight: 70.0kDa), which increased the proliferative activity of macrophages by 40.5% compared with the substrate lily polysaccharide.

Embodiment 2

[0017] Lily polysaccharides were modified with complex enzymes according to the following conditions and operations: substrate lily polysaccharide concentration 35 / L; β-mannanase concentration 600U / L, β-glucanase 300U / L; pH 6.0 (PBS buffer liquid); temperature 45°C; time 40min. After the reaction, heat the solution to 100°C, keep it warm for 5min, then centrifuge (6000rpm) to remove the enzyme protein, put the supernatant into a dialysis bag (molecular weight cut-off: 5.0kDa) and dialyze (4°C, 12h), after dialysis The sample was centrifuged (5000rpm), and the supernatant was freeze-dried to obtain a white powder lily polysaccharide modified product (molecular weight: 50.3kDa), which increased the proliferative activity of macrophages by 45.2% compared with the substrate lily polysaccharide.

Embodiment 3

[0019] According to the following conditions and operations, the complex enzymes were used to modify lily polysaccharides: substrate lily polysaccharide concentration 40 / L; β-mannanase concentration 700U / L, β-glucanase 350U / L; pH 6.5 (PBS buffer liquid); temperature 45°C; time 50min. After the reaction, heat the solution to 100°C, keep it warm for 5min, then centrifuge (6000rpm) to remove the enzyme protein, put the supernatant into a dialysis bag (molecular weight cut-off: 5.0kDa) and dialyze (4°C, 12h), after dialysis Centrifuge the sample (5000rpm), take the supernatant and freeze-dry to obtain the modified product of lily polysaccharide (molecular weight: 40.5kDa) in the form of white powder. Compared with the substrate lily polysaccharide, the proliferative activity of macrophages is increased by 55.0%.

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Abstract

The invention relates to an organism modified preparation method for improving the activity of lily polysaccharides, belonging to the field of agricultural-product functional ingredient preparation technologies. Lily polysaccharides (403.3kDa) are subjected to organism modification (namely enzyme-method controllable degradation) by adopting a compound enzyme (beta-mannanase and beta-glucanase) method. After the modification, the lily polysaccharides are subjected to separation and purification, and the molecular weight of product lily polysaccharides is 8.0-70.0kDa. The in-vitro immunoregulation activity of the lily polysaccharides and modified products of the lily polysaccharides is subjected to model research by adopting in-vitro macrophages RAW264.7. The activity of the modified lily polysaccharides is improved remarkably, and the macrophage proliferation activity is increased by 40.5-77.2% compared with a substrate. The activity of the lily polysaccharides is relatively low no matter the molecular weight of the lily polysaccharides is too high or too low. The method for improving the immunocompetence of the lily polysaccharides is created, and the biologically modified lily polysaccharides have great application values in the fields of medicines and functional foods.

Description

technical field [0001] The invention belongs to the technical field of preparation of functional components of agricultural products, and relates to a biomodification preparation method for improving the activity of lily polysaccharide. Background technique [0002] Lily is one of the first batch of medicine and food approved by the Ministry of Health. It not only has a wide range of clinical applications, but also has a good health care effect. Lily polysaccharide is one of the main functional factors of lily. Studies have shown that lily polysaccharides have biological activities such as enhancing immunity and anti-tumor. Through ultrasonic-assisted hot water extraction, we extracted a lily polysaccharide from lily, with a molecular weight of 403.3kDa, mainly composed of mannose and glucose in a molar ratio of 1:1 through β-1,4 glycosidic bonds (containing a small amount of 1 →2 or 1→6 bond type), also contains a small amount of arabinose and a small amount of sulfate gro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/14C12P19/04
Inventor 陈志刚张丹妮王芬郭晓玉
Owner NANJING AGRICULTURAL UNIVERSITY
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