JPS5836393A - Preparation of abscisic acid by fermentation method - Google Patents

Abstract

PURPOSE:To collect the titled substance industrially and advantageously from a culture, by cultivating a microorganism having the ability to produce abscisic acid in a culture medium containing a guanine compound in a specific amount or more. CONSTITUTION:A microorganism, e.g. Cercospora rosicola (FERM-P No.6128), having the ability to produce abscisic acid is cultivated in a culture medium containing 5mug/ml or more guanine compound, e.g. guanine, guanosine or 5'-guanylic acid, a carbon source, e.g. glucose or ethanol, nitrogen source, e.g. ammonia, and Na, K, etc. at 2-9 pH for 5-20 days by the spinner culture method with aeration to accumulate a considerable amount of the abscisic acid in the culture.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing abscisic acid by fermentation. In detail, in the production of abssaicic acid, a microorganism capable of producing abscisic acid is cultured in a culture medium, abssaicic acid is accumulated in the culture, and abssaicic acid is collected from the culture. The present invention relates to a method for increasing the amount of abscisic acid produced by causing the presence of a guanine compound. The objective is to provide an industrially advantageous abscise/#1 method. Regarding the production of abscisic acid by microorganisms, Cercospora rosicola (
c@rcoaport rosicola), but the amount produced is still low and unsatisfactory.

That is, in the production of abscisic acid by baking soda,
It is known that several tens of micrograms of absisic acid accumulates in a medium containing natural nutrients such as yeast extract, shoulder extract, malt extract, etc., but almost no absisic acid is produced in a medium containing peptone. (gxp@rie
wtia 33 rxsss (tsyy)). One of the reasons why the amount of absisic acid produced is so small is that OJ
The above natural nutrients added as medium components contain both substances that promote the production of abscisic acid and substances that inhibit it, and these substances act in a way that contradicts each other. This is thought to limit the production of abscisic acid. Furthermore, the above-mentioned natural nutrients are tetravalent substances and are not necessarily suitable for use in industrial fermentation production of absisic acid.

Considering this situation, the present inventor searched for a substance that promotes the production of absisic acid by absisic acid-producing bacteria, and found that the production of absisic acid was significantly promoted by adding guanine P and its derivatives to the culture solution. I found that - to be done.

Until now, it has not been known that guanine compounds promote the production of abscisic acid, and the present invention is based on this new finding.

Examples of the guanine compounds related to the present invention include guanine, guanosine e 5'-guanylic acid, sl-guanylic acid, 51-guanosine triphosphate, and other guanine haze conductors. These guanine compounds can be used alone or in combination, and the amount added to the medium is 5 to 500 μl in terms of guanine. Furthermore, these O-guanine compounds can be used in both synthetic and natural media, and in combination with other nucleic acid-related compounds such as adenine, cytosine, xanthine, hyposanthin, uracil, thymine, or derivatives thereof. can be used.

As the microorganisms used in the present invention 1111, any microorganisms having the ability to produce abscisic acid can be used. Is it Cercospora rosico as a specific strain? An example of this is H3 Koko Kensō Yori No. 6128.

Carbon sources for the culture medium used by Motoya INK include glucose,
7 Tuctose, galactose, lactose, shakerose, innose, trehalose, sebiose, atubinose, lactose and natural products containing them (
For example, blackstrap molasses, potato extract, lees @), ethanol and others, 1iQL lactic acid, pyruvic acid, succinic acid, other organic acids, glutamic acid, lysine, asbagic acid, arginine or methionine 7, g')syn,
Amino acids such as valine, leucine, and isoleucine can be used alone or in combination.

Ammonia is a nitrogen source added to the culture medium.

Urea, ammonium sulfate, ammonium chloride.

Inorganic and organic ammonium compounds such as ammonium acetate, ammonium citrate, ammonium phosphate, ammonium carbonate, glutamic acid, lysine, aspartic acid, asparagine, proline.

Essential amino acids such as arginine, corn/steve/liquor, meat extract, sake lees extract, soybean meal decomposition product.

Peptone, hydrolysis of various fermented substances - 2 various protein-containing substances, etc. can be used. Some of these natural nitrogen sources are difficult to use as guanine-based compounds.

Inorganic substances include Na+ K+ Mytt, ca
, Col Ni + Zn, Cu, Fs, Cj
, PO418041Not and the like can be used.

Other vitamins are added to the medium.

For culture, the oxygen transfer rate in the medium is 0.01 threat
The culture is carried out by shaking culture or aerated agitation culture at a temperature of /d or more.

p)I during culture is preferably 2 to 9. Culture g# temperature is 1
A temperature of 8 to 30°C is suitable. As the pH neutralizing agent, ammonia water, caustic soda, calcium hydroxide, calcium carbonate, magnecum phosphate, urea, etc. are used. The cultivation period is 5-20 days, and significant amounts of absisic acid accumulate in the culture.

Examples of the present invention are shown below.

Example 1 ゜100-Todoroki j) 14jF potato dextrose prosthe (product of Difoo company. This 2.4Ii contains 2II
(contains glucose and potato 201/equivalent extract)
Botetope xistrose agar medium (I) H4LS) containing Raiten 2II was placed in a large test tube (160 × 16
fin) and sterilized under pressure at 120°C for 15 minutes to prepare an agar slant medium according to a conventional method. After the agar solidified, Cercospora rosicote No. 6128 was inoculated over the entire surface of the medium and cultured at 25° C. for 20 days.

Dispense 8- sterilized water onto the culture on the agar slant and
The microbial cells are suspended in this using a glass rod, and the resulting suspension of microbial cells is used as a seed culture. This seed culture (L5-) was inoculated into fermentation medium 10114 containing guanine at a concentration of seed^, which was dispensed into large test tubes (160 smxiam), and cultured with shaking in a test tube shaker for 15 days.

Table 1 (埒/-) (μm17m) No additive @ Gourd 1 1.0 γcL3 25 81450
97.67.5 1
0&2 1 ao 11 a3 2&0 10&7 5CL0 121.4 10(LO1112 15(LO11!4i) The accumulated amount of ab 1 sicidic acid corresponding to each guanine concentration is shown in Table 1. The basic composition is as follows: boteto dextrose syrup (D
ifco) 14jl/dJ, pHa! ! Example λ Instead of guanine K, 1 ml of guanosine 93d.

The relaxation was carried out in the same manner as in Example 1, except that 1201 di/- of 5'-guanylic acid was added to the medium. The results are shown in Table 2.

Table 2 Additive-free 648 Guanokun (93) 87.6

Claims (1)

[Claims] Cultivating microorganisms capable of producing abscisic acid in a medium,
A method for producing abssaicic acid in which abssaicic acid is accumulated in a culture and abssaicic acid is collected from the culture, the fermentation being characterized in that a guanine-based compound is present in the culture medium in an amount of 5 g/'sJ or more as guanine. Method for producing abscisic acid by method.