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Chlorella-based metallothionein preparation method

A metallothionein and chlorella technology, which is applied in the preparation methods of metallothionein and peptides, and methods based on microorganisms, etc., can solve the problems of high transfer fee, large equipment investment, low output, etc., to improve quality and reduce Production cost, rapid growth effect

Inactive Publication Date: 2015-04-01
TIANKE HUIYANG TECH DEV TIANJIN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, rabbit liver is used to produce metallothionein in China. The investment in equipment is large, the transfer fee is high, and the output is low. It is difficult to meet the current market demand for metallothionein.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Chlorella-based metallothionein preparation method, according to the following steps:

[0028] Step 1, the raw material of chlorella culture fluid is the NaHCO of 2g / L 3 , 0.5g / L of KNO 3 , 0.02g / L of KH 2 PO 4 , 0.3mg / L of V B1 and a V of 1.0μg / L B12 , put the culture solution in the reactor, sterilize at 120°C, inoculate the chlorella and the culture solution at a ratio of 1:3 by volume, then place it in a light incubator at 25°C, cultivate it for 12 hours, and then put the reaction Place the device in an incubator at 25°C without light, incubate for 5 hours, and then put it in an incubator with light for 12 hours, repeat this 10 times, and add 5 μmol / L zinc sulfate to the chlorella in the logarithmic growth phase Solution, after culturing for 120 hours, add 5 μmol / L copper chloride solution to the chlorella after the above-mentioned treatment, and then cultivate for 10 days;

[0029] Step 2, after culturing for 10 days, concentrate the culture solution and the ...

Embodiment 2

[0035] Chlorella-based metallothionein preparation method, according to the following steps:

[0036] Step 1, the raw material of chlorella culture fluid is the NaHCO of 2g / L 3 , 0.5g / L of KNO 3 , 0.02g / L of KH 2 PO 4 , 0.3mg / L of V B1 and a V of 1.0μg / L B12 , put the culture solution in the reactor, sterilize at 120°C, inoculate the chlorella and the culture solution at a ratio of 1:1 by volume, then place it in a light incubator at 25°C, cultivate it for 12 hours, and then put the reaction Place the device in an incubator without light at 25°C for 5 hours, and then put it in an incubator with light for 12 hours. Repeat this three times. Add 100 μmol / L chloride to the chlorella in the logarithmic growth phase. Zinc and zinc gluconate mixed solution, after cultivating for 24 hours, then add 100 μmol / L copper sulfate and copper gluconate mixed solution to the chlorella after the above treatment, and then cultivate for 3 days;

[0037] Step 2. After 3 days of cultivation, ...

Embodiment 3

[0043] Chlorella-based metallothionein preparation method, according to the following steps:

[0044] Step 1, the raw material of chlorella culture fluid is the NaHCO of 2g / L 3 , 0.5g / L of KNO 3 , 0.02g / L of KH 2 PO 4 , 0.3mg / L of V B1 and a V of 1.0μg / L B12 , put the culture solution in the reactor, sterilize it at 120°C, inoculate the chlorella and the culture solution according to the volume ratio of 1:2, then put it in the light incubator at 25°C, cultivate it for 12h, and then put the reaction Place the device in an incubator without light at 25°C for 5 hours, and then put it in an incubator with light for 12 hours. Repeat this 8 times. Add 50 μmol / L chloride to the chlorella in the logarithmic growth phase. Zinc solution, after cultivating for 90 hours, add 70 μmol / L copper chloride solution to the chlorella after the above treatment, and then cultivate for 8 days;

[0045] Step 2, after culturing for 8 days, concentrate the culture solution and the chlorella in it...

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Abstract

The invention provides a chlorella-based metallothionein preparation method. The method comprises the steps: respectively feeding zinc and copper ions to a chlorella reactor for culturing; concentrating through a microfiltration membrane; breaking the wall of chlorella at the normal pressure; extracting chlorophyll; and extracting metallothionein. According to the preparation method, algae can be used as the source of metallothionein, so that the production cost of metallothionein is obviously decreased; metallothionein extracted through the chlorella has outstanding advantages, can grow fast, and is high in accumulation and easily processed; dry chlorella powder with metallothionein extracted out can be used for preparing a feed of the animal husbandry; chlorophyll in the chlorella is extracted through ethyl alcohol, so that the influence of chlorophyll on extraction and separation of metallothionein can be reduced, and the quality of metallothionein can be improved; and then zinc and copper are utilized for alternative stress induction for preparing metallothionein; and therefore, the productive rate of the obtained metallothionein is up to 10g / 250kg, and the high-purity metallothionein can be obtained after purifying.

Description

technical field [0001] The invention relates to the technical field of metallothionein preparation, and more specifically relates to a method for preparing metallothionein based on chlorella. Background technique [0002] Metallothionein is a metal-binding protein produced by microorganisms, animals and plants. It is a short peptide rich in cysteine ​​and has a high affinity for various heavy metals. It is a low molecular weight protein with a very high content of cysteine ​​residues and metals. The metals combined with it are mainly cadmium, copper and zinc, which widely exist in various organisms from microorganisms to humans, and their structures are highly conserved. [0003] According to the application and development of metallothionein in the three fields of cosmetics, health products and new medicines, combined with the actual level of consumers in my country, as well as the steps and speed of new medicine development, the annual demand of the cosmetics industry is ...

Claims

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Application Information

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IPC IPC(8): C12P21/02C07K14/825C07K1/14C07D487/22A23K1/00C12R1/89
Inventor 季延滨李涛
Owner TIANKE HUIYANG TECH DEV TIANJIN CO LTD
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