JPH03280891A - Manufacturing method of nicotinic acid amide using microorganisms - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a method for producing nicotinamide using microorganisms. More specifically, the present invention relates to a method for producing nicotinic acid amide using a microorganism, in which 3-cyanopyridine is subjected to a hydration reaction by the action of a specific genus of microorganism, and the resulting nicotinic acid amide is separated and obtained. Nicotinamide is a water-soluble vitamin B
It is one of the group and is known to be a component of coenzymes. Furthermore, nicotinic acid amide derivatives are used in pharmaceuticals and the like.

[Conventional technology and problems]

Conventionally, the method for producing nicotinamide has been a chemical synthesis method in which 3-cyanopyridine obtained by ammoxidation of β-picoline is catalytically hydrated using a reduced copper catalyst.
-22710 and Japanese Unexamined Patent Publication No. 50-111077) are known.

However, this method has problems such as the production of by-products and a complicated purification process, and there has been a desire to develop a new and industrially advantageous manufacturing method.

On the other hand, methods using microorganisms of the genus Brevibacterium (see Japanese Patent Publication No. 62-21519), Rhodoconcus Method using microorganisms of the genus Rhodoculus
-470 Publication, Appl, Enν1ron, Mi
crobiol, 541766-1769 (198
8)) are known. The production method using these microorganisms is considered to be a more industrially advantageous production method than chemical production methods because it can react under mild conditions and produces fewer reaction by-products. However, with regard to these microorganisms, there are examples of experiments in which nicotinic acid amide was produced using extremely large numbers of bacterial cells, and there are only basic experiments that have been conducted on these microorganisms. It has not yet been reached.

[Means for solving problems]

The present inventors have diligently searched for new and more effective microorganisms for the purpose of industrial production, and as a result, among the strains provided, we found that the genus Cellulomonas or Streptomyces IIE
The present invention was completed by discovering a microorganism belonging to the genus that has high 3-cyanopyridine hydration activity.

That is, the present invention provides a method for producing nicotinic acid amide in which 3-cyanopyridine is converted into nicotinic acid amide by a hydration reaction caused by the action of a microorganism, in which the microorganism is of the genus Cellulo+l1onas or the genus Streptomyces. This is a method for producing nicotinic acid amide using a microorganism, which is characterized by using a microorganism belonging to the nitrile group and having the ability to hydrate the nitrile.

Examples of microorganisms used in the present invention include Streptomyces griseus.
iseus) IFo 3355, Cellulomonas fimi (Cellul-omonas fist) IPo
12107 etc. can be mentioned.

As a medium for culturing these microorganisms, a medium in which an amide or nitrile compound is added to a medium in which these microorganisms can grow is usually used.

For example, carbon sources include glucose, sucrose, glycerol, etc., nitrogen sources include peptone, meat extract, yeast extract, amino acids, various inorganic and organic acid ammonium salts, other inorganic salts, trace metal salts, vitamins, etc., as necessary. A medium to which an amide or nitrile compound is added is used.

The amide and nitrile compounds to be added include aliphatic compounds such as isobutyronitrile, isobutyramide, propionitrile, propionamide, methacrylonitrile, and methacrylamide; aromatic compounds such as benzonitrile and benzamide;
Examples include heterocyclic compounds such as 3-cyanopyridine and nicotinic acid amide.

The culture is carried out aerobically at a pH of 6 to 10, preferably 7 to 9, and a temperature of 15 to 37°C, preferably 25 to 30°C.
Do this for ~7 days.

In the present invention, as a method for producing nicotinic acid amide from 3-cyanopyridine, for example, 3 - A method of adding cyanopyridine, adding a substrate to a suspension of bacterial cells fixed by a method of adding cyanopyridine (e.g., crushed bacterial cells, crude enzyme, purified enzyme, etc.) or a suspension of bacterial cells or treated bacterial cells, etc. There are methods such as a method in which a substrate is added to the culture solution during culturing of microorganisms and a reaction is carried out simultaneously with the culturing.

Although the substrate concentration in the reaction solution is not particularly limited, 1
The range of mM to IM is preferable, and the substrate can be added to the reaction solution all at once or in portions. The reaction temperature is preferably 5 to 50°C and the pH is preferably 4 to 10. Although the reaction time varies depending on the substrate concentration, bacterial cell concentration, and other reaction conditions, it is generally sufficient to complete the reaction within 10 minutes to 48 hours.

Nicotinic acid amide can be recovered from the reaction solution by a conventional method. For example, bacterial cells can be removed from the reaction solution by centrifugation or the like, the reaction solution can be concentrated to dryness, and then recovered and purified by recrystallization or the like.

[Experiment example]

Next, the present invention will be explained by examples, but the present invention is not limited only to these examples.

In addition, nicotinic acid amide was quantified by high performance liquid chromatography (using a Showa Denko 5hodex F-411A column).

Example 1 Polypeptone 0.5χ, yeast extract 0.2χ, glucose 0.5χ, KJPOa O,05L KHtPOa
O, 05χ, Mg5Oa-7) 1200.05χ, 0.
042mM CoCIg, 0.042+nM Fe50
．． ．． 0.042mM N1Cb, 0.042a+M M
nCIz, 0.042+sM Cu5Oa, 0.042
mM Zn5Oa, 0.042mM NaffiMoO
a, Streptomyces griseus (S
treptomyces griseus) IFO33
Washed bacterial cells (2.0 mg) prepared by culturing 55 with shaking at 30°C for 48 hours were added to 0.5 d of 50 mM potassium phosphate buffer (pH 7) containing 10 wM3-cyanopyridine.
, 2) and reacted at 25°C for 26 hours.

After the reaction was completed, the bacterial cells were removed by centrifugation and the supernatant was examined, and the production of 0.09 mM nicotinic acid amide was observed.

Example 2 Polypeptone O, SZ, yeast extract 0.2χ, glucose 0.5L KJPOn O,05χ, KHtPOa
O,05χ, Mg5O.

-78100,05χ, 0.04211M CoC
1g, 0.042mM Fe1o4.0.0421
1M N1Ch, 0.042mM MnCIg,
0.042+gM Cu5Oa, 0.042mM
Zn5Oa, 0.042mM NaJoOn, 0.2χ
Cellulomonas fimi (Cellulomona fimi) was grown in a medium containing isobutyronitrile and 0.2χ isobutyramide.
s fimi) IFO12107 at 30°C
Washed bacterial cells (2.0 mg) prepared by shaking culture for 1 hour
0,5 d of 50-H with 0 mM 3-cyanopyridine
Suspended in potassium phosphate buffer (pH 7,2) and heated at 25°C.
After 26 hours of reaction, the bacterial cells were removed by centrifugation and the supernatant was examined, and the production of 0.05 mM nicotinic acid amide was observed.

Claims (1)

[Claims] In the method for producing nicotinic acid amide in which 3-cyanopyridine is converted to nicotinic acid amide by a hydration reaction caused by the action of a microorganism, Cellulomonas (Cellulomonas) is used as the microorganism.
ulomonas) or Streptomyces (St
1. A method for producing nicotinic acid amide using a microorganism, the method comprising using a microorganism belonging to the genus Reptomyces and having the ability to hydrate the nitrile.