Method for producing d-alpha-tocopherol through enzyme engineering method

A production method and technology for tocopherol, which are applied in the field of enzymatic engineering transformation to produce d-α-tocopherol, can solve the problems of limited output growth, rising prices, inability to meet market demands, etc., so as to reduce costs and improve output and quality. Effect

Inactive Publication Date: 2014-11-05
上海粮心谷业生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because the growth of its output is limited by the source of raw materials, it cannot meet the market demand, and it is in a seller's market. The price has been rising year by year, and it is currently more than 10 times higher than that of synthetic vitamin E.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Utilize Synechocystis sp. PCC6803 to prepare γ-tocopherol methyltransferase:

[0033] A. First take 2.2L of DMEM-1 culture solution, sterilize it at 121°C for 10 minutes, then add 2 grams of Synechocystis sp. nourish;

[0034] B. Injecting 30 milliliters of citric acid with a molar concentration of 25% into the culture solution, and centrifuging the culture solution after the acid treatment, 4°C, 5000rpm, 30 minutes, using the hydrated substance obtained after the separation Wash with pure water several times to obtain the cleaned bacteria;

[0035] C. Finally, use Mulitibeadsshocker B300 to oscillate, oscillating frequency: 60 times / min, amplitude: 20MM, after oscillating for 1 hour, break up the bacteria, and centrifuge to obtain the precipitate;

[0036] D, put the above precipitate into 30% ethanol solution as extraction solvent, after stirring at 20°C for 30 minutes, carry out centrifugation, 4°C, 5000rpm, 30 minutes, then remove the ethanol solution ...

Embodiment 2

[0039] Example 2. Preparation of γ-tocopherol methyltransferase by Synechococcus sp. MBIC10224

[0040] A. Add 1% by mass of Synechococcus MBIC10224 wet cells to a 3L cell generator containing 2.2 liters of DMEM-1 culture medium, and cultivate for 28 hours at 30°C under anaerobic conditions with a stirring speed of 300rpm ;

[0041] B, when cultivating, add 30 milliliters of 25% citric acid solutions in the nutrient solution, collect the 180 gram wet objects that bacterium body obtains by centrifugation culture and wash 3 times with water, obtain and clean cell body;

[0042] C. Use Mulitibeadsshocker B300 to oscillate, stir for 1 hour, break the cell body, and centrifuge to obtain the precipitate;

[0043] D, add water to 180 grams of sediment, after making it recover slurry, obtain dry cell body by spray drying;

[0044] E. Add 1000 milliliters of ethanol as an extraction solvent to the dried cell body, stir at 50° C. for 30 minutes, then centrifuge to remove the ethanol s...

Embodiment 3

[0047] Example 3. Preparation of d-alpha-tocopherol:

[0048] (1) Take 100g of mixed tocopherols, heat in a water bath at 50-75°C, and add 200g of 90% ethanol solution under stirring.

[0049] (2) Add 2% trimethylamine to react for 2 hours, then add 2 ml of γ-tocopheryl methyltransferase solution to carry out conversion for 12-24 hours.

[0050] (3) After extraction, washing, drying, concentration and molecular distillation, d-α-tocopherol is obtained.

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Abstract

The invention relates to an industrialized method for producing d-alpha-tocopherol through an enzyme engineering method. According to the method, d-beta, d-gamma, d-delta tocopherols are converted into the d-alpha-tocopherol through gamma-tocopherol methyl transferring enzyme, so as to be different from the conversion of the chemically synthesized metal catalyst methyl. The method is high in efficiency, is pollution-free and is used for producing the real pure natural d-alpha-tocopherol product, namely, pure dextrorotation alpha-tocopherol.

Description

technical field [0001] The invention relates to a d-alpha-tocopherol production technology, in particular to a method for producing d-alpha-tocopherol through enzyme engineering transformation, which belongs to the technical category of biological products. Background technique [0002] Vitamin E (VE) is an essential component of human nutrition. It has various physiological functions such as promoting energy metabolism of the human body, preventing vascular sclerosis, protecting the liver, regulating gonad function, delaying aging and inhibiting free radicals. Effective drugs play an important role in anti-sterility, anti-cancer, anti-aging, treatment of coronary heart disease, myocardial infarction and thrombus, arteriosclerosis, increase the body's immunity, improve the utilization of oxygen in the blood and muscle endurance, etc., can be used In many fields such as pharmaceutical industry, food industry, cosmetics industry, etc. [0003] Vitamin E is a general term for ...

Claims

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Application Information

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Patent Type & AuthorityPatents(China)
IPC IPC(8): C12P17/06C12N9/10C12R1/89
Inventor熊军娄彩玲李国兴陈芝荣单晓昌
Owner上海粮心谷业生物科技有限公司