JPS5836397A - Preparation of s-adenosyl methionine - Google Patents

Abstract

PURPOSE:To prepare the titled substance, by cultivating a yeast, belonging to the genus Saccharomyces, Candida or Hansenula, and having the ability to produce the titled substance, in a culture medium containing a lower peptide constituted of glycine and/or alanine. CONSTITUTION:A yeast, e.g. Saccharomyces cerevisiae IFO 2346, belonging to the genus Saccharomyces, Candida or Hansenula, and having the ability to produce S-adenosyl methionine is cultivated in a culture medium containing the methionine, glycine, alanine or a lower peptide constituted of the glycine and alanine. The lower peptide is added to the culture medium according to the conventional method in a concentration of 0.02g/dl or more, and the cultivation is carried out under aerobic conditions at 3-8 pH for 2-10 days. The resultant S- adenosyl methionine is then collected from the microbial cell and/or culture fluid.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing 8-adenoflumethionine (hereinafter referred to as RAM) by a fermentation method, and more specifically, it relates to a method for producing 8-adenoflumethionine (hereinafter referred to as RAM) by a fermentation method. The present invention relates to a method of culturing yeast in a liquid medium to efficiently produce 8 μM Mt.

111M is an important substance involved in the production of fat, protein, and other substances in living organisms. Recently, it has been found to be effective in treating 8 mu mk liver hemoptysis, hyperlipidemia, arteriosclerosis, depressive illness, neurological manifestations of psychosis, degenerative threshold syndrome, neurological disorders, insomnia, etc. has been found9,
Mass production is expected.

Conventionally, as the 81M01111 method, various culturing methods using fungal organisms have been known, but among them, the method using yeast is considered to be preferable.
w A method using genus Ises, Candy f, genus O fungi (91, for example, Jowr+a turtle1of11a*tari@1
6gy, Ma.z 121a, p. 247 (1975), etc.), genus Vichia, genus Dotorz, genus Cryptococcus,
A method using microorganisms of the genus Ha/Trichosporon, Frechella, Hanseniaspotus, SporoSat, and Ribo (41 meetings)
1i52-17118), etc. However, even in these O methods, the amount of 8AM accumulated is not necessarily fully satisfactory, and further improvements have been sought.

Therefore, the present inventors conducted extensive studies on improving the amount of SAM accumulated using the mil method, and found that it is effective to use a medium supplemented with a certain type of lower peptide, methionine/tomoK, which is a precursor of SAM. discovered this and completed the present invention.

In other words, the purpose of the present invention is to produce 8
An object of the present invention is to provide a method for producing AM, and an object of the present invention is to provide a method for producing 13AM by culturing yeast capable of producing 8muM in a methionine-containing medium. is achieved by adding to the medium a lower peptide consisting of Arani/.

The yeast used in the present invention may be any yeast as long as it has the ability to accumulate flAM in a methionine-containing medium, but yeasts belonging to the genus Saccharomyces are preferred, and specific examples include Naccalomyces cerevisiae po 2546, Candida Φ ! Sedonye/Sis IFO0960, Ha/Zenuts Fabianii IP0 1
570 etc.'. tlI - Even these natural and artificial mutant bacteria can be used in the same manner as Kagishii, which has the ability to write BAM. It is essential to add lower peptides consisting of glycine and alanine. ・Lower peptides here refer to dipeptides, tripeptides, and tetrapeptides. Specific examples of such lower peptides include: For example, glycylglycine, glyculglycylglycine, glycylglycylglycine, D, L-Atzel D, L-Acunino, L-Atzunil L
-Afnyl L-72, L-alanyl L-alanyl L
- Alanyl L-792, L-72 Nylglycine, etc. - These O lower peptides are K(L
It is added at a concentration of O2#/jJ or more, especially cLo41
/dl or higher is appropriate.Other conditions for the culture medium used in the present invention may be determined according to conventional methods. Acids, hydrocarbons, alcohols such as methanol and ethanol, etc. can be used. Further, in the culture medium, ordinary nitrogen sources, inorganic salts, and organic micronutrients are used as necessary.For O 2 culture, aerobic conditions are preferable, and the culture temperature is preferably in the range of 20[11' to 40C]. During culture, adjusting the pH of the medium to a range of 3 to 8 will yield the most desirable results.Thus, after 2 to 10 days of culture, 8 mu. Since M is generated and accumulated, paAMt* can be obtained by processing it according to a conventional method.
After extraction at ice-cooling and neutralization with carbonated water bulk liquor,
If necessary, contact with strongly acidic cation exchange@fat, then 5Aiit-moulding, then elute with sulfuric acid, add Su/Ta/Guste/#1 to the eluate to precipitate 8mM. The present invention will be described in more detail with examples below, but the present invention is not limited thereto. After the cultivation was completed, the bacterial cells and the culture solution were separated by centrifugation, and the bacterial cells were diluted with approximately 5 times the volume of 1
．． After extraction with 5M perchloric acid, the resulting extraction supernatant was mixed with the culture solution, and then subjected to beno-J chromatography (developing solvent: ethanol/1-butanol/water/acetic acid/sodium 1-pyrophosphate). Water II wave-55/1G/I
A spot corresponding to 8AM was detected using an ultraviolet detector, and this was extracted with 11M hydrochloric acid, and 8AMjlt- in the sample was detected from the absorbance of 2604 times.

Run 11i11 Glucose 5 lid J, Polypeptone [1L
5t/di, K H4F Os a41 /d J,
1cmMP Oa'(L41/dJ, MgS, 0
4-7H! 0α021/d1% yeast extract CL21/
d1. Agar slant medium (PH
1 platinum loop of Natsucalomyces cerevisiae IP'02546 grown for 2 days on 60) was mixed with 10 sucrose.
1/di.

Yeast extract 1t7ds, KH, PO2α41/d
i%Mg804・7H20α011/dj, urea (separate play) LSI/di.

L-methionine Q, 751/d j, Oa c, t
2 @ 2 a2゜α (J21/dj, Zo80a ・7
H20α25vml/dl, Fe3O4・7H20α2
5m1/lN, MnSO4・4~6T120 t25
wsl/di, 0u804” 5 H2O2μl/d
i, HmBOs 2μI/dl.

0oCI2 ・6 H2O0,2fil/di, II
1 fil/di and a predetermined amount of glycylglycyl/karana 5 PHl adjusted to 5 d of heat-sterilized medium,
Shake at 28C for 5 days, determine the accumulated amount tm of 8M using the 90 method, and write the result as #! Table 1 shows Table 90. From this result, it is recommended to add glycylglycine.
It can be seen that the amount of DaA)C) produced can be improved.

Cultivation was carried out under the same conditions as in Example 1, except that the lower peptides shown in the K111g2 table were added to the medium at α11/Ill 09 degrees instead of K111g2. Second result
Shown in the table.

j[Table 2 *1 It was added to the culture medium without heating at room temperature without heating.

Example 5 Cultivation was carried out under the same conditions as in Example 1, except that the strains shown in Table 3 were used for Natsucharomyces cerevisiae (P0 2346) and the lower peptides shown in Table 5 were added to the medium. . The results are shown in Table 5.〇Table 5

Claims (1)

[Claims] tS-adenosylmethioni/Ikudo #! In the method for producing S-adenosylmethionine by culturing yeast having methionine in a methionine-containing medium, a lower peptide consisting of glycine and alanine is added to the medium.
A method for producing 10 Jll of 8-adenosylmethioe as Ili mold. 2. A patent claim aS that belongs to the genus Ha/Deida and the genus I1m1
The method described in Section 1 of S.