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Method for extracting cardamine hirsute plant selenium protein

A technology of plant selenoprotein and extraction method, which is applied in the processing field of rice chestnuts, can solve problems such as complex procedures, unfavorable development and utilization of plant organic selenium, and inability to effectively extract plant selenoprotein, so as to achieve improved extraction efficiency and simple process Effect

Inactive Publication Date: 2018-09-04
安徽过湾农业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing extraction technology is not only complicated in process, but also unable to effectively extract the plant selenoproteins in the broken rice chestnuts, which is not conducive to the development and utilization of plant organic selenium by researchers. Therefore, we propose an extraction of plant selenoproteins in broken rice chestnuts method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] First, carry out preliminary extraction: take 200g fresh leaves of Cardamine in the flask, add 800ml sample extract solution that is pre-cooled, homogenize with a high-speed tissue homogenizer (intermittently, keep low temperature), keep the temperature at 4℃, and rotate at 6000r / min Centrifuge for 30 minutes, take the supernatant, and accurately measure the volume; then salt out: under slow stirring, slowly add (NH 4 ) 2 SO 4 , Make it reach 35% saturation, then keep it at 4℃ for 60min, homogenize with a high-speed tissue homogenizer at 6000r / min and centrifuge for 30min, discard the precipitate, accurately measure the volume of the supernatant, and stir slowly in the supernatant. Add slowly (NH 4 ) 2 SO 4 , Make it reach 55% saturation, keep it at 4℃ for 1h, then centrifuge at 4℃, 6000r / min for 30min, discard the supernatant; finally desalting: dissolve the precipitate with column equilibration buffer, and load on SephadexG-25 column with column equilibration The buffer...

Embodiment 2

[0022] In the first embodiment, the following steps are added:

[0023] The composition of the sample extract in step (1) is 20mmol / L PH7.8 Tris-HCL buffer, 1mmol / LEDTA, 10mmol / L MgCL 2 , 10mmol / L NaHCO 3 , 10mmol / L β-mercaptoethanol, 1% PVP.

[0024] First, carry out preliminary extraction: take 200g fresh leaves of Cardamine in the flask, add 800ml sample extract solution that is pre-cooled, homogenize with a high-speed tissue homogenizer (intermittently, keep low temperature), keep the temperature at 4℃, and rotate at 6000r / min Centrifuge for 30 minutes, take the supernatant, and accurately measure the volume. The composition of the sample extract is 20mmol / L PH7.8 Tris-HCL buffer, 1mmol / L EDTA, 10mmol / L MgCL 2 , 10mmol / L NaHCO 3 , 10mmol / L β-mercaptoethanol, 1% PVP; then salting out: under slow stirring, slowly add (NH 4 ) 2 SO 4 , Make it reach 35% saturation, then keep it at 4℃ for 60min, homogenize with a high-speed tissue homogenizer at 6000r / min and centrifuge for 30min, di...

Embodiment 3

[0026] In the second embodiment, the following steps are added:

[0027] Step (2) maintains the pH of 7.8 during the salting-out process, and adjusts the pH value with ammonia or sulfuric acid.

[0028] First, carry out preliminary extraction: take 200g fresh leaves of Cardamine in the flask, add 800ml sample extract solution that is pre-cooled, homogenize with a high-speed tissue homogenizer (intermittently, keep low temperature), keep the temperature at 4℃, and rotate at 6000r / min Centrifuge for 30 minutes, take the supernatant, and accurately measure the volume. The composition of the sample extract is 20mmol / L PH7.8 Tris-HCL buffer, 1mmol / L EDTA, 10mmol / L MgCL 2 , 10mmol / L NaHCO 3 , 10mmol / L β-mercaptoethanol, 1% PVP; then salting out: under slow stirring, slowly add (NH 4 ) 2 SO 4 , Make it reach 35% saturation, then keep it at 4℃ for 60min, homogenize with a high-speed tissue homogenizer at 6000r / min and centrifuge for 30min, discard the precipitate, accurately measure the vol...

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Abstract

The invention discloses a method for extracting a cardamine hirsute plant selenium protein. The method comprises the following steps: performing primary extraction, namely putting 200g of fresh leavesof cardamine hupingshanesis into 800ml of a precooled sample extract, homogenizing (intermittently, at a low temperature) by using a high-speed tissue homogenizer, keeping the temperature at 4 DEG C,centrifuging for 30 minutes at a rotation speed of 6000r / minute, collecting supernate, and measuring a volume accurately; performing salt separation, namely with slow stirring, slowly putting (NH4)2SO4 into the supernate till a saturation degree of 35%, keeping the temperature of 4 DEG C, leaving to stand for 60 minutes, homogenizing by using the high-speed tissue homogenizer at a speed of 6000r / minute, centrifuging for 30 minutes, and abandoning precipitate. By adopting the method, the purpose of extracting a plant selenium protein can be achieved, the method has the characteristics of beingsimple in process and high in extraction efficiency, convenience can be brought to scientific operators to utilize the plant selenium protein in cardamine hupingshanesis, and the problems that not only is a conventional extraction technique complex in procedure, but also the plant selenium protein in the cardamine hupingshanesis cannot be effectively extracted, and organic plant selenium cannot be well developed and utilized by scientific operators, can be solved.

Description

Technical field [0001] The invention relates to the technical field of cardamine processing, in particular to a method for extracting cardamine plant selenoprotein. Background technique [0002] Huping Cardamine, a cruciferous plant, was first discovered on Huping Mountain in Hunan in 1983, hence its name. Cardamine Huping is a perennial herb with obvious underground stems. The stems stand up or bend. The upper part often has branches, 20~2500cm high, and the whole plant is smooth and hairless. Single leaf alternate, papery, 4~325cm long, 5~14cm wide, kidney-shaped or nearly heart-shaped, palm-shaped veins, with serrated edges, basal leaves occasionally 1 to 4 pairs of leaflets: petiole length 2.5~3.0cm, With inconspicuous wings. It is basically enlarged and shaped like leaf ears without holding stems. The racemes are terminal or axillary, without bracts. The calyx is ovate, 5-6mm long and 3-4mm wide; petals are white with reticulate veins. Broadly obovate. 8-10mm long, 5-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14C07K1/30C07K1/16
CPCC07K1/145C07K1/16C07K1/30
Inventor 樊浪生樊高俊
Owner 安徽过湾农业科技有限公司
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