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 improve output and quality and reduce costs. Effect

Active Publication Date: 2012-12-26
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. Preparation of γ-tocopherol methyltransferase by Synechocystis sp. PCC6803:

[0033] A. First, take 2.2L of DMEM-1 culture solution, sterilize it at 121°C for 10 minutes, then add 2g of Synechocystis PCC6803, make it at 28°C and rotate at 300rpm for 48 hours. nourish;

[0034] B. Inject 30 ml of citric acid with a molar concentration of 25% into the culture solution, and carry out centrifugation treatment to the culture solution after adding acid, at 4° C., 5000 rpm, 30 minutes, and the hydrate obtained after the separation is treated with Washed with pure water for several times to obtain washed bacterial bodies;

[0035] C. Finally, use Multibeadsshocker B300 to oscillate, oscillating frequency: 60 times / min amplitude: 20MM, after 1 hour of oscillating, the bacteria are broken, and the precipitate is obtained by centrifugation;

[0036] D, put the above-mentioned precipitate into 30% ethanol solution as extraction solvent, after stirring at 20 ° C for 30...

Embodiment 2

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

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

[0041] B, when cultivating finishes, add 30 milliliters of 25% citric acid solution to the culture solution, collect the 180 grams of wet body obtained by the centrifugal separation culture, and wash the 180 grams of wet body with water 3 times to obtain the washed cell body;

[0042] C, use Multibeadsshocker B300 to shake, after stirring for 1 hour, the cell body is broken, and centrifugation is performed to obtain a precipitate;

[0043] D. Add water to 180 grams of sediment to restore the slurry, and obtain dry cell bodies by spray drying;

[0044] E. Add 1000 ml of ethanol as an extraction solvent to the dry cell body, stir at 50° C. for 30 minutes, and then cent...

Embodiment 3

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

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

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

[0050] (3) After extraction, water 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 technology for producing d-α-tocopherol, in particular to a method for producing d-α-tocopherol by enzymatic engineering transformation, which belongs to the category of biological product technology. Background technique [0002] Vitamin E (VE) is an essential component of human nutrition. It has various physiological functions such as promoting human energy metabolism, preventing vascular sclerosis, protecting liver, regulating gonadal 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 thrombosis, arteriosclerosis, increase immunity, improve the utilization rate of oxygen in the blood and muscle endurance, etc. In the pharmaceutical industry, food industry, cosmetics industry and other fields. [0003] Vitamin E is a general term for tocopherol compounds. It is not only a c...

Claims

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Application Information

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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