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Method for converting 3-cyanopyridine into nicotinic acid by using gibberella intermedia CA3-1

A technology of cyanopyridine and gibberella, which is applied in the biological field and can solve problems such as long half-life

Active Publication Date: 2012-06-13
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a method for converting 3-cyanopyridine into nicotinic acid by using the Gibberella CA3-1 strain in view of the technical difficulties and existing problems in the existing nicotinic acid production method. high conversion efficiency, long half-life, etc.

Method used

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  • Method for converting 3-cyanopyridine into nicotinic acid by using gibberella intermedia CA3-1
  • Method for converting 3-cyanopyridine into nicotinic acid by using gibberella intermedia CA3-1
  • Method for converting 3-cyanopyridine into nicotinic acid by using gibberella intermedia CA3-1

Examples

Experimental program
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Effect test

Embodiment 1

[0016] Embodiment 1 Utilizes Gibberella CA3-1 to convert 3-cyanopyridine into nicotinic acid

[0017] (1) Preparation of cell liquid culture of Gibberella

[0018] Pick a ring of Gibberella strains on the solid Chapei culture medium, inoculate it in a 250mL Erlenmeyer flask containing 30mL seed medium, and culture it on a shaker at 200r / min at 30°C for 20-24h to Logarithmic phase, that is, the cell liquid culture of the strain.

[0019] (2) The composition and proportioning of the fermentation medium are:

[0020] Carbon source 50-100g / L; organic nitrogen source 10-30g / L; NH 4 Cl 20-30g / L; K 2 HPO 4 0.1-1.0g / L; MgSO 4 0.1-1g / L; CuCl 2 0.01-0.1g / L; ZnSO 4 0.01-0.10g / L; MnSO 4 0.01-0.10g / L; FeSO 4 ·7H 2 O 0.01-0.10g / L; pH 6.5-7.5, 121°C

[0021] Sterilize under high pressure steam for 20min. The carbon source is selected from one or both of glucose, sucrose and maltose; the organic nitrogen source is selected from one or two of white corn steep liquor, peptone a...

Embodiment 2

[0028] Embodiment 2 Studies on thermal stability of Gibberella CA3-1

[0029] Take an appropriate amount of resting cell suspension, place them in water baths at 30°C, 40°C, and 50°C respectively, take samples regularly, convert 3-cyanopyridine according to the method described in Example 1, and use HPLC to track and detect the substrate The consumption and the production of the product are compared with the initial enzyme activity to calculate the relative enzyme activity, and the logarithm of the residual enzyme activity (lnRA) is plotted against time, and the slope K Decat That is, it represents the loss rate of enzyme activity, the results are shown in figure 2 .

Embodiment 3

[0030] The substrate spectrum research of embodiment 3 Gibberella CA3-1

[0031] Get the resting cell of appropriate amount of Gibberella, add different nitrile compounds (final concentration 20mM) respectively, transform by the method in embodiment 2, the concentration of HPLC method detection substrate and product, calculate relative enzyme activity and conversion rate, The results are shown in the table below.

[0032] Table 1

[0033]

[0034]

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Abstract

The invention belongs to the field of biotechnology, and particularly relates to a method for converting 3-cyanopyridine into nicotinic acid by using gibberella intermedia CA3-1. The strain is collected at the China General Microbiological Culture Collection Center with a collection number of CGMCC No. 4903. The strain can be used for converting the3-cyanopyridine into the nicotinic acid under the condition of substrate concentration of 5.2 g / L (50 mM), the substrate conversion efficiency reaches up to 98 percent; under the condition of the optimum reaction temperature of 30 DEG C, the half life reaches up to 231.1 h; and the nitrilase of the strain CA3-1 has a broad substrate spectrum, and various single nitrile and double nitrile compounds can be converted.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for converting 3-cyanopyridine into nicotinic acid by using a strain of Gibberella intermedia CA3-1. Background technique [0002] Nicotinic acid is 3-pyridinecarboxylic acid, also known as nicotinic acid, white odorless crystal, non-toxic, slightly soluble in water, easily soluble in organic solvents, and is a vitamin with simple structure and stable physical and chemical properties. Niacin, as an essential chemical substance for the human body and animals, participates in the redox process of tissues, and has the functions of promoting cell metabolism and dilating blood vessels. Nicotinic acid is also an important pharmaceutical raw material and chemical intermediate, and has been widely used as food additives, feed additives, pharmaceutical intermediates, reactive dyes, etc. With the development of the economy and the improvement of people's living standards,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/12C12R1/645
Inventor 许正宏吴燕史劲松龚劲松李恒李会陆震鸣朱小燕
Owner JIANGNAN UNIV
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