Preparation technology of niacinamide

A production process, nicotinamide technology, applied in the field of nicotinamide production technology, can solve the problems of low nicotinamide product quality, complex refining process, low product concentration, etc., to achieve increased product concentration and production intensity, high product concentration, The effect of high enzyme activity

Active Publication Date: 2018-04-17
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the production of nicotinamide by wild bacteria often produces by-product nicotinic acid, resulting in low quality nicotinamide products;
[0005] In addition, the current biological prod

Method used

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  • Preparation technology of niacinamide
  • Preparation technology of niacinamide
  • Preparation technology of niacinamide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1 Bacterial classification and the mensuration of enzymatic activity

[0050] (1) Strain construction

[0051] The strain used in this example is: E. coli BL21(DE3) / pET-30a(+)-NHaseP, a genetically engineered strain expressing nitrile hydratase derived from Bordetella petrii DSM12804. For the specific method of genetically engineered bacteria construction, please refer to the invention patent application document with the application publication number CN104498466A entitled "Nitrile Hydratase and Its Application". The base sequence of the nitrile hydratase gene in the genetically engineered bacteria is shown in SEQ ID NO.1.

[0052] Genetically engineered bacteria E.coli BL21(DE3) / pET-30a(+)-pENHase-1229 expressing nitrile hydratase derived from Aurantimonas manganoxydans SI859A (Aurantimonas manganoxydans SI859A), the base of nitrile hydratase gene in genetically engineered bacteria The base sequence is shown in SEQ ID NO.2. The construction method of gen...

Embodiment 2

[0073] Example 2 Production of Nicotinamide Using Genetically Engineered Bacteria E.coli BL21(DE3) / pET-30a(+)-NHaseP

[0074] (1) Strain activation: use an inoculation loop to take the genetically engineered strain E.coliBL21(DE3) / pET expressing nitrile hydratase derived from Bordetella petrii DSM12804 (Bordetella petrii DSM 12804) preserved in a -80°C glycerol tube in the strain tube -30a(+)-NHaseP strains were drawn on the surface of LB-Kan-agar solid medium (plate or eggplant bottle), and the plate was placed upside down in a constant temperature incubator at 37°C for 12 hours;

[0075] (2) Seed cultivation: re-inoculate the activated strains into 40mL of primary seed culture medium, culture with shaking at 37°C for 12 hours to obtain primary seed liquid; then, transfer the primary seed liquid into 400mL of secondary seed culture medium with shaking at 35°C for 4 hours to obtain a secondary seed liquid;

[0076] Among them, the primary seed medium is LB medium, 10g / L pepto...

Embodiment 3

[0090] Example 3 Production of Nicotinamide by Genetically Engineered Bacteria E.coli BL21(DE3) / pET-30a(+)-pENHase-1229

[0091] (1) Strain activation: use an inoculation loop to take the genetically engineered strain E.coli BL21 (DE3) expressing nitrile hydratase derived from Aurantimonas manganoxydans SI859A (Aurantimonas manganoxydans SI859A) preserved in a -80°C glycerol tube in the strain tube / pET-30a(+)-pENHase-1229 strains were drawn on the surface of LB-Kan-agar solid medium (plate or eggplant bottle), and the plate was placed upside down in a constant temperature incubator at 37°C for 12 hours;

[0092] (2) Seed cultivation: re-inoculate the activated strains into 50 mL of primary seed culture medium, shake and culture at 37°C for 12 hours to obtain primary seed liquid; then, transfer the primary seed liquid into 500 mL of secondary seed culture medium with shaking at 35°C for 4 hours to obtain a secondary seed liquid;

[0093] Among them, the primary seed medium is...

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Abstract

The invention discloses a preparation technology of niacinamide. The preparation technology comprises the following steps of building of gene engineering bacteria, fermenting culture of engineering bacteria, and performing catalytic reaction, wherein the engineering bacteria comprise escherichia coli host cells and a nitrile hydratase gene of which the base sequence is shown in SEQ ID NO.1 or SEQID NO.2; in the catalyzing process, pure water is used as a reaction medium to prepare a cell suspension liquid; 3-cyanopyridine is added into the cell suspension liquid by batches, and the concentration of the 3-cyanopyridine in the reaction system is controlled to be 90g/L or less, so as to obtain the niacinamide. The preparation technology has the advantages that the gene engineering bacteria containing the recombinant nitrile hydratase with high enzyme activity is used for fermenting culture, the pure water is used as the reaction medium, and the 3-cyanopyridine is added into the cell suspension liquid in a batched base matter adding way, so as to perform the hydration reaction at high efficiency, thereby obtaining the niacinamide product; the conversion rate of the hydration reactionis more than 99.9%, the quality of the obtained product is high, and the impurities and byproducts are not produced.

Description

technical field [0001] The invention belongs to the field of biochemical industry, in particular to a production process of nicotinamide. Background technique [0002] Nicotinamide, also known as Nicotinamide, English name: Nicotinamide, Nicotinamide, also known as vitamin B3, belongs to the B vitamins (Vb), is the biological body of nicotinamide adenine dinucleotide (coenzyme I) and nicotinamide adenine dinuclear A component of glycoside phosphate (coenzyme II), involved in the metabolism of carbohydrates, fats and proteins. As a vitamin, nicotinamide has the effect of anti-rough skin disease, and has unique curative effect on certain skin diseases and neurological disorders. The largest application field of nicotinamide is as a feed additive, and it can also be added to food to supplement the vitamins needed by the human body. Studies have found that adding nicotinamide or nicotinic acid to feed can improve the disease resistance and growth rate of livestock and poultry....

Claims

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Application Information

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IPC IPC(8): C12P17/12C12N15/70C12N15/60C12R1/19
CPCC12N9/88C12N15/70C12P17/12C12Y402/01084
Inventor 杨立荣周海胜吴坚平徐刚
Owner ZHEJIANG UNIV
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