JPS6336B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention involves culturing Penicillium chrysogenum in a medium containing taratannin (hereinafter referred to as the first step).
The present invention relates to a new and improved method for producing gallic acid, which is characterized in that the culture is incubated at 37 to 60° C. (hereinafter this step is referred to as the second step). The production of gallic acid by culturing Penicillium chrysogenum is described in Japanese Patent Publication No. 47-41550. This method involves culturing Penicillium chrysogenum in a medium containing taradannin to accumulate tannase, and the action of this tannase decomposes tartannin into gallic acid and quinic acid to isolate gallic acid. This is the manufacturing method. However, with this conventional method, it takes a long time to completely decompose cod tannin, and when the medium is sterilized at high temperature, the polymer substances in cod powder elute and become highly viscous, making it difficult to cultivate Penicillium chrysogenum in this medium. In this case, the multiplied bacterial cells and the polymeric substance form a slurry, which hinders post-processing such as separating the bacterial cells or purifying gallic acid, and this method is not satisfactory as an industrial method for producing gallic acid. Nakatsuta. Therefore, the present inventors conducted various studies to establish an industrially advantageous method for producing gallic acid, and as a result, they focused on the fact that the culture conditions for Penicillium chrysogenum and the optimal operating conditions for tannase are different. The present invention was completed based on the knowledge that gallic acid could be produced simply and efficiently by combining the two methods. The purpose of the first step is to cultivate Penicillium chrysogenum on a solid medium containing taradannin to accumulate tannase while degrading taradannin, and the purpose of the second step is to decompose undegraded taratannin and/or additional ingredients as will be explained later. The purpose is to complete the decomposition of tartannin. The first step is to mix Taratannin with a carrier, and add a solution of salts such as magnesium sulfate, phosphate, urea, ammonium chloride, and ammonium sulfate to this mixture until the moisture content
This can be done by spraying at a concentration of 40 to 60%, sterilizing it, and then adding Penicillium chrysogenum seed koji to solid culture. Tara tannin may be used in the form of cod (Caesalpina spinosa) powder or its extract. Examples of carriers include silicates such as diatomaceous earth and perlite, rice husks, sawdust or mixtures thereof. The amount of carrier used varies depending on the type of carrier and tartannin, but it is used to the extent that caking or agglomeration does not occur during sterilization or culturing.
For example, when cod powder is used as cod tannin, a carrier that weighs 25 to 100% of the cod powder is usually used. As Penicillium chrysogenum, for example, the WH-3 strain (deposit number 499) deposited with the Institute of Fine Technology is preferably used. 1st
The culture in the process is carried out under the optimum growth conditions for Penicillium chrysogenum as shown in Table 1, i.e.
It is preferable to carry out the reaction at 37°C, especially about 30°C, for about 24 hours.

【table】

[Table] The second step is carried out by incubating the culture from the first step at 37-50°C. The second step should be carried out under conditions in which the tannase activity is exhibited most strongly and is utilized efficiently. For this purpose, for example, incubation is performed at the optimum pH and temperature for tannase, or additional cod tannin is supplemented before incubation to increase the utilization rate of tannase. In addition, as shown in Tables 2 and 3, the activity of tannase is 37 to 50℃, especially 40 to 45℃.
It is most strongly exerted at pH 5 to 8, especially pH 6.5 to 7.5.

【table】

【table】

[Table] As is clear from the above description, the most preferred embodiment of the present invention is to grow Penicillium chrysogenum WH-3 strain in a solid medium containing taratannin.
The culture is cultured at 30°C for about 24 hours, and the culture is further supplemented with taratannin and incubated at 40-45°C at pH 6.5-7.5. The gallic acid thus produced can be isolated by extracting the reaction product of the second step with hot water and further recrystallizing it. For hot water extraction, it is industrially advantageous to carry out the second step in a column and then to pass hot water through this column. Next, the present invention will be explained in more detail with reference to Examples. Note that the yield in each Example was calculated from the yield of gallic acid from cod tannin in cod powder or its extract. Example 1 20 g of cod powder containing 7.8 g of cod tannin, 5 g of perlite, and 10 g of rice hulls in a 300 ml beaker
Take this, add 0.8g of urea, 0.24g of monopotassium phosphate,
Dipotassium phosphate 0.24g, magnesium sulfate 0.04g
After adding 15 ml of solution containing 100
Sterilize for 30 min at °C. After cooling, 20 ml of 2.8% sodium hydroxide aqueous solution was sprinkled on the Penicillium, which was then cultured for 5 days on bran to which cod powder was added.
Add 1 g of seeds of Chrysogenum WH-3 strain,
Mix well and culture at 30℃ for 24 hours to obtain 6g (yield 77.2
%) of gallic acid is obtained. This culture was then incubated at 45°C for 24 hours to yield 6.7g (85.3% yield).
67 g of a reaction product containing 50% gallic acid was obtained. Example 2 0.1N to 20g of culture obtained in the same manner as Example 1
- Add NaOH solution and fill into a column with a diameter of 20 mm. This was incubated at 45℃ for 24 hours and 2.3g (yield
96.3%) of gallic acid is obtained. Pass 3 times the amount of hot water through the column (flow rate SV = 1) and add 2.2g
An extract containing gallic acid (yield 93%) is obtained. The maximum extraction concentration was 5.8%. Example 3 The yield of gallic acid when the conditions of cultivation in the first step and incubation in the second step were varied were obtained, and the following results were obtained.

[Table] Example 4 Various amounts (varying from 0 to 5.2 g) of cod tannin obtained by extraction of cod powder were added to a culture containing 1.0 g of gallic acid obtained in the same manner as in Example 1, PH
6.5, and incubated at 45°C for 40 hours to obtain the results shown in the following table.

[Table] Yield in additive plot - Yield in control plot * Increase rate (%) =

Claims (1)

[Scope of Claims] 1. Penicillium chrysogenum is cultured in a solid medium containing Taratannin, and then the culture is
A method for producing gallic acid characterized by incubation at 50℃. 2. The production method according to claim 1, wherein the Penicillium chrysogenum is WH-3 strain. 3. The production method according to claim 1 or 2, wherein the culture temperature is 5 to 37°C. 4. The manufacturing method according to claim 1, wherein the incubation is carried out at a pH of 5 to 8. 5. The manufacturing method according to claim 1 or 4, which further comprises adding cod tannin and then incubating it. 6. The production method according to claim 1 or 5, using cod powder or an extract thereof as cod tannin. 7 Penicillium chrysogenum WH-3 strain was incubated at approximately 30°C for 24 hours in a medium containing cod powder or its extract.
The production method according to any one of claims 1 to 6, which comprises culturing for about 30 minutes, adding cod powder or an extract thereof to the culture, and incubating at 40 to 45°C at pH 6.5 to 7.5. .