JPS6193116A - Antibacterial antiphage - Google Patents

Abstract

PURPOSE:An antibacterial and antiphage agent that is composed of a specific compound, thus being used to prevent phage infection in fermentation industry, because it has a stronger antiphage activity than ascorbic acid. CONSTITUTION:A compound of the formula (R is H, 1-3C lower alkyl; n is integer of 0-2) is used as an antibacterial and antiphage agent. It has the same high level of antibacterial activity and higher antiphage activity than ascorbic acid. It is used, as an antiphage agent, in the fermentation industry such as aminoacid fermentation, nucleic acid fermentation, or organic acid fermentation. Further, the addition of a divalent transition metal, preferably CuSO4 or CuCl2 to the compound of the formula increases the antibacterial activity.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an antibacterial and antiphage agent, and more particularly to an antibacterial and antiphage agent comprising a compound of the following formula (r).

It is known that L-ascorbic acid, ie, vitamin C, exhibits antibacterial activity in the presence of divalent transition metals, and also exhibits antibacterial activity itself.

While conducting research on L-ascorbic acid derivatives and related compounds thereof, the present inventors discovered that certain derivatives themselves have antibacterial and antiphage activity, leading to the completion of the present invention.

That is, the gist of the present invention is represented by the formula: %Formula %) [In the formula, R represents hydrogen or a lower alkyl group having 1 to 3 carbon atoms, and n represents an integer of O to 3. ] The antibacterial and antiphage agent consists of the compound shown below.

Among the compounds (1), the derivatives in which n is 0, their physical properties, and synthesis methods are as follows: Carbohydrate RCs
68.313-319 (1979),
Other derivatives (1,) can be synthesized according to the synthesis method.

Compound (1) used in the present invention has high antibacterial activity equivalent to that of ascorbic acid, but when a salt of a divalent transition metal is present, the activity is enhanced or the antibacterial spectrum is expanded.

Preferred examples of divalent transition metal salts include sulfates, nitrates, and halides (e.g., chlorides and bromides) of copper, iron, manganese, zinc, etc., and among them, copper salts such as CuSO4, Cu(NC)+)2, CuCL
, CLIBr2 and the like are particularly preferred.

When a salt of a divalent transition metal is coexisting, the amount added is as follows:
It can be determined as appropriate depending on the type and concentration of the derivative (D), the type of bacteria, etc.

Furthermore, the compound (D) of the present invention has stronger antiphage activity than ascorbic acid. Therefore, it is extremely useful in fields where it is necessary to prevent damage caused by phage infection, such as in the fermentation industry.

Examples of fermentations in which the compound (1) of the present invention can be used as an antiphage agent include L-lysine, L-glutamic acid,
- Amino acid fermentation such as arginine, nucleic acid fermentation such as wild boar, guanosine, inosinic acid, organic acid fermentation such as citric acid and lactic acid, production fermentation of enzymes such as amylase and protease, solvent fermentation such as acetone butanol fermentation, antibiotic fermentation, etc. can be mentioned. The amount added to the medium is
Although it varies depending on the type of fermentation, the type of microorganism used, the type of compound (1), etc., generally at least IO-'M
, preferably 10-'M, e.g. 10-' to 10-3
It is sufficient to select M. Next, a manufacturing example is shown,
The method for producing compound (1) will be illustrated.

Production Example 1 5.6-dideoxy-D-glycero-hex-2,3:
5. Preparation of 6-genonol 1,4-lactone 10.0 g of neat ascorbic acid was dissolved in a 30% acetic acid solution of hydrogen bromide 5
After stirring at room temperature overnight, the solvent was distilled off under reduced pressure. Next, 20 ml of toluene was added, and the solvent was distilled off again under reduced pressure. The residual brown syrupy product was dissolved in water/acetic acid (50ml/1150ml) and added with zinc powder (30ml).
g was gradually added over 1 hour. After stirring for an additional hour, it was filtered and the filtrate was distilled under reduced pressure. The residue was redissolved in water, passed through an ion exchange resin (SK-IB), water removed under reduced pressure, and dissolved in Ethacool/ethyl acetate (20 ml).
/20ml) was added, and the solvent was further distilled off. The resulting brown powder was recrystallized from ethyl acetate/chloroform to obtain 5.0 g of the title compound as pale yellow crystals. Melting point 147-14
9°C0 C013C-N: 173.47 (1-C), 117
゜50 (1-C), 157.42 (3-C), 78.3
5 (4-C), 131.64 (5-C), 123.48
(6-C) ppm0 Example 1 5.6-sideoxy-D-glycerol hexa-2,3:
5.6-phenono-1,4-lactone and phage were combined and tested for antiphage activity as follows.

The phages were added to 0.02M Tris-HCl buffer (pH 7°4).
Add 1 to 4XI to 07 PFU/ml, mix the compound solution thereto to a concentration of 10-'M, and add 37
Incubated at ℃ for 30 minutes. After incubation,
Take a part of the solution and cool it with water to dilute it (1g of NaσC, Mg
SO4-7H, 00, 25g, gelatin 0.03g, 0
．． The reaction was stopped by diluting 100 times or more with 01M phosphate buffer (pH 7,0) LO.

The number of phages before and after the reaction was measured using the usual plaque counting method (
M. H. Adama, “Bacteriophag.
es”Interscience Publish
rs (New York, USA), and the phage survival rate was calculated to evaluate the antiphage activity of the compound.

The results are shown in Table 1.

Example 2 5,6-sideoxy-D-glycerol hexa-2,3;
5.6-Genonor 1,4-lactone was tested for antibacterial activity in combination with bacteria as follows.

The test bacteria were added to 0.02M Tris-HCl buffer (pH 7.4) at a concentration of 1 to 4 x 10' cel Is/ml, and the compound solution was mixed at a concentration of 1X10-3M. It was then incubated at 37°C for 60 minutes in the absence or presence of 1X10-6M Cu2-.

After that, take a part of the solution and add diluted solution (NaC (! 1 g
.

MgSO4・7Ht00.25g, gelatin 0.03g
10.01M + J7 acid buffer (pH 7,0) 1 (liter)
The reaction was stopped by diluting the mixture to 00 times or more.

The number of colonies before and after the reaction was measured by a conventional method, and the residual rate of bacteria was calculated to evaluate the antibacterial activity of the compound. Note that Cu'- in IxlO-6M alone had no antibacterial activity.

The results are shown in Table 2. The numbers in parentheses are the results in the presence of Cu sulfur.

In the table, the bacterial abbreviations represent the following bacteria.

Claims (1)

[Claims] 1. Formulas: ▲ Numerical formulas, chemical formulas, tables, etc.▼ [In the formula, R represents hydrogen or a lower alkyl group having 1 to 3 carbon atoms, and n represents an integer of 0 to 2. An antibacterial antiphage agent consisting of the compound shown in ].