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Method for preparing S-adenomethionine

A technology of adenosylmethionine and methionine, which is applied in the field of preparation of S-adenosylmethionine, can solve the problems of increasing the difficulty of separation process, long fermentation period, cumbersome process and the like, and achieves simplified operation, easy regeneration and good enzyme stability. Effect

Inactive Publication Date: 2011-11-30
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The fermentation method can achieve a higher SAM yield, but this method has certain defects: the fermentation cycle is too long, and after the fermentation is over, SAM needs to be extracted from the cell, the process is cumbersome, and it is easy to introduce other substances in the cell. Impurities, which increase the difficulty of the subsequent separation process
This method needs to synthesize ATP respectively and extract a large amount of SAM synthetase from the bacterial strain producing SAM synthetase so as to meet the needs of synthesizing SAM, and the operation is more complicated [Shanghai Guojia Biochemical Engineering Technology Research Center Co., Ltd., a new adenosine A In vitro enzymatic conversion method of thionine [P], China: CN 1850980A, 2006.10.25]

Method used

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  • Method for preparing S-adenomethionine
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Examples

Experimental program
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Effect test

Embodiment 1

[0029] Yeast medium: glucose 40g / L, urea 2.0g / L, potassium dihydrogen phosphate 1.5g / L, magnesium sulfate heptahydrate 0.5g / L, zinc sulfate heptahydrate 4.0×10 -3 g / L, ferrous sulfate heptahydrate 3.0×10 -3 g / L, manganese chloride tetrahydrate 0.3×10 -3 g / L, anhydrous calcium chloride 1.0×10 -3 g / L, biotin 0.05×10 -3 g / L.

[0030] The inoculum amount of yeast was 10%, cultured on a shaker at 120 rpm at 30°C for 24 hours, and centrifuged at 4000 rpm for 20 minutes. Take the yeast paste and store it at -7°C for later use.

Embodiment 2

[0031] Example 2: Using glucose as an energy donor, using AMP to synthesize SAM.

[0032] Take a 500ml shake flask, prepare 49mM AMP, 0.14M glucose, 100g Saccharomyces cerevisiae mud, 0.22M potassium dihydrogen phosphate, 157.5mM MgCl 2 , 16.5mM MgSO 4 , 5mM NH 4 Cl, 40.2mM L-Met, 4.5mL toluene, 35mM mannitol, 10mM xylitol, and 300mL reaction liquid, adjust the pH to 6.8 with sodium hydroxide, and stir at 33°C for 4 hours at a low speed. , precipitated with 40% trichloroacetic acid, quantitatively analyzed SAM by HPLC, and contained SAM 4.5g / L in the conversion solution.

Embodiment 3

[0033] Example 3: Using glucose as an energy donor to synthesize SAM using AR.

[0034] Take a 500ml shake flask, prepare 45mM adenosine (AR), 0.2M glucose, 120g air-dried Candida parapsilosis, 0.17M potassium dihydrogen phosphate, 0.044M dipotassium hydrogen phosphate, 210mM ​​MgCl 2 , 3.7mM NH 4 Cl, 40mM L-Met, 30mM glycerol, 54mM mannitol and 300ml of reaction liquid, adjusted pH to 7.0 with potassium hydroxide, stirred at 37°C for 8h at low speed, after the reaction, precipitated with 40% trichloroacetic acid , carry out quantitative analysis to SAM by HPLC, contain SAM 6.5g / L in the conversion liquid.

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Abstract

The invention discloses a method of preparing S-adenosyl methionine, according to the method, by using polyhydroxyl organic reagent to keep the activity of enzyme, by taking adenosine triphosphate precursor, L-methionine and phosphate ions as a substrate and taking glucose and / or Maltose as an energy supplier, and by adding the combination of metal ions, the producing strain with permeability can be used for producing S-adenosyl methionine in the way of whole-cell biocatalysis. The invention can use the combination of metal ions to regulate the metabolism flow to improve energy self-coupling efficiency; and organic reagent is added to keep the activities of thalli and enzyme. The strain with permeability is used for producing S-adenosyl methionine, thereby shortening the synthesis time and accumulating outcomes outside cells, so that the cost of the following separation can be reduced, and the operational steps also can be simplified.

Description

technical field [0001] The present invention relates to a method for preparing S-adenosylmethionine (SAM), and more particularly relates to a method for preparing S-adenosylmethionine by a coupled enzymatic conversion method. Background technique [0002] As we all know, S-adenosyl-L-methionine (S-adenosyl-L-methionine, SAM) is an important metabolic intermediate widely present in animals, plants and microorganisms, and is also an important physiologically active substance in the human body. The effects of transmethylation, transsulfurization and transaminopropylation have good curative effects on arthritis, depression, liver dysfunction, pancreatitis, etc. SAM is also a high-level healthcare drug for cancer prevention, cardiovascular disease and anti-aging. In 1999, the US FDA approved SAM as a health product, and quickly became one of the best-selling nutritional products in the United States. With the improvement of people's quality of life and the renewal of health con...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/32C12R1/13C12R1/645C12R1/84C12R1/72
Inventor 应汉杰周雪文张磊柏建新李振江欧阳平凯
Owner NANJING TECH UNIV
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