JPS5828296A - Preparation of s-adenosylmethionine - Google Patents

Abstract

PURPOSE:To prepare the titled substance, in high efficiency, by culturing yeast belonging to Saccharomyces genus, Candida genus, etc. in a methionine-containing liquid medium added with non-sulfur type amino acid or its salt. CONSTITUTION:Yeast belonging to Saccharomyces genus, Candida genus or Hansenula genus and capable of producing S-adenosylmethionine, e.g. Saccharomyces cerevisiae IFO2346, etc. is cultured in a methionine-containing medium added with a non-sulfur type amino acid devoid of sulfur atom in the molecule, e.g. glycine, methylglycine, L-alanine, L-leucine, L-aspartic acid, L-lysine, L-tryptophan, etc. at a concentration of >=0.02g/ml. The cultivation is carried out by conventional method under aerobic conditions, and the S-adenosylmethionine produced and accumulated in the microbial cell and/or cultivation liquid is separated therefrom.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for enriching 8-adenosylmethionine (hereinafter abbreviated as 8AM) by the fIII# method, and more specifically, to a method for producing 8-adenosylmethionine (hereinafter abbreviated as 8AM) using a methionine-containing liquid containing a non-sulfur amino acid or a salt thereof. The present invention relates to a method for cultivating yeast in a medium and efficiently producing 8-musume irises.

8M is a substitute for fats, proteins, sugars, etc. in the body.
It is an important substance involved in
It has been found that it has a therapeutic effect on hepatemia, hyperlipidemia, kinetosclerosis, depression and neurological forms of psychosis, degenerative arthritis, neurological pain sensation, insomnia, etc., and its mass production. It is expected that 〇Conventional 1. Cultivation methods using various microorganisms are known as methods for producing aAM, but among them, methods using yeast are said to be preferred, and a specific example thereof is a method using microorganisms of the genus Satucharomyces or Ca/Deida. (
For example, Journal ofBaet・r4G1GBy
, MaOX p. 121.267 (1975), etc.), Bichia, Rhodotorula, Cryptococcus j1% Hansenula II, ) Lycosboro, Frechella, Ha/Xeniasbo, Sboroborozes, Ribo (Ses) is a method using the fungal organism τ of the genus Debaryomyces (Special Publication No. 52
-17118) are known. however,
Even in these methods, the SAMc) accumulation type cannot necessarily be said to be sufficiently efficient, and further improvements have been desired.

Therefore, as a result of intensive investigation into the accumulation of SAM by the ai#al method, Kinei #4J et al.
They found that using I5r was effective and completed the present invention.

That is, the purpose of the present invention is to produce H using a fermentation method with excellent productivity.
The purpose of the present invention is to provide a method for producing AM (Q), and an object of the present invention is to provide a method for producing BAM by culturing yeast capable of producing BAM in a menaonin-containing medium, in which sulfur atoms are added to the molecule. This can be achieved by adding a non-sulfur-based a) acid or its salt to the medium.

The #mother used in the present invention may be any mother as long as it has the ability to accumulate 8M in a methionine-containing medium, but it is not limited to
c-c, arushi cmon)X, gear/dig (,9an
(Ca) f4, Hansenula (HaΩS・Ω-1-) #mother belonging to M is preferable, and its specific examples include Satucharomyces cerevisiae F0 2546, Candida machidoniae/Sisue Fl'0 0960. Hansenula Fabianii El'0 1570 etc. are being criticized. 'Also, these natural and artificial mutations-
It can be used in the same way as a gagiri with M production capacity.

In the present invention, it is essential to add a salt of the non-sulfur amino acid to the menaonin-containing medium. The non-sulfur amino acids referred to here refer to amino rI&, which does not contain a sulfur atom in the molecule, and its examples include glycine/, methylglycine, L-alanine, L-leucine, and L-alanine.
-isoloidino, D, L-valine, β-alanino, D
, L-α-amino-ni-m#, n1L-β-7mi/-
ri-am, L-serine, L-threonine, L-homoserine, r-amino-β-hydroxy-Mgl! Neutral aliphatic amines such as L-asparagi/[
%L-glutamic acid etc. 1! Acidic aliphatic amino acids; L-arginine, L-lysine, L-ornithine,
L-asparagia, basic aliphatic amino acids such as L-glutamine; L-proline, L-hysteresis/
, hydroxy-L-proline, and the like; aromatic amino acids such as L-7 enylalanine, L-telosif, L-tritophane, and the like. Examples of these salts include alkali metal salts and salts. These non-sulfur amino acids are usually added to the culture medium at a concentration of [1Q211/C1t or higher,
In particular, it is appropriate to set the concentration to 0-041/d1 or more.

Other conditions for the culture medium used in the present invention may be determined according to conventional methods, including glucose, 7-ucrose, 79-ctose, and other shields, organic acids such as acetic acid, hydrocarbons, methanol, and ethanol. In addition, a normal nitrogen source, no rr
a-salts, organic micronutrients are used as needed.

The culture is carried out under good aerobic conditions, and the culture temperature is 2υC・ra40
A range of C is preferred. When culturing, adjust the pH of the medium from 6 to 8.
The most desirable results will usually be obtained if the temperature is within the range of .

Thus, after 2 to 10 days of culture, E3AM is produced and accumulated in the cells and/or the culture solution, and RAM can be obtained by processing this in a conventional manner.

For example, EIAM monomers are extracted with perchloric acid, the extract is neutralized with potassium bicarbonate under water cooling, and then brought into contact with a strong cationic father-converting resin as required, and then 8 μM Mi
After absorbing the layer, it is eluted with sulfuric acid, and SAM can be isolated by adding phosphotungstair #lt to the effluent to precipitate it.

The present invention will be described in more detail below with reference to examples of implementation. However, the present invention is not limited thereto. In addition, the constant violet of BAM in the example is as follows, -u
I went. In other words, after culturing is completed, the m
*The fraction was separated and extracted with a 15M perchlorine solution containing approximately 50% of the total gold. Then, after mixing with the obtained extraction supernatant solution T-hair culture solution, paper coutonography (Example 8 Bath medium: ethanol/1-butanol/water/vinegar acupuncture/1% sodium pyrophosphate aqueous solution-55150150/1 /1), a spot of the SAM phase was detected with an ultraviolet edge detector, extracted with α1M hydrochloric acid, and the amount of 511M in the sample was detected from its 260% fi absorbance.

Example 1 / h Course 51 / (11, Horihef) N a51
/lit.

KH2PO4 (lL41/dl, KH2O4CL41
/dt, Mg804.7 H20α11/dl% #Mother extract 0.21/(tl, yIA Dog) 1 platinum loop of Satucharomyces cerevisiae FO2546 grown for 2 days on an agar slant medium (pIito) consisting of 217 dl was mixed with 1017 dl of Nucrose.

#Mother extract 11/d1%KH2PO4L417QLe,
MgBua ・7 Hz OQ, 011/dz, urine;
1c (not listed separately) t51/dl.

L-methio/(J, 751/dl%a*ct2・2H
200-021/dl, ZnBOa ・7H20CL
25tsl/dl, FeSO4・7) (20α25m
1/lt1%MnSO44~6 Hz0 t25a&
I/de%Cu1304・5H202μl/dl, H
4BOs2μl/≦it, a ・)v12 ・6
H20α2 μl/dt, XI 1 μkl/d
PHAO consists of 4 te and a predetermined amount of glycino, and 14 te,
The cells were placed in heat-sterilized medium 5d and shaken at 28C for 5 days. The SAM accumulation i1 was then quantified and the results are shown in Table 1.

Table 1 From this result, SAMO can be
It can be seen that the amount of raw g can be improved.

Example 2 Instead of Glyshi/1'amino sales shown in @2 table, α11/
Real m except for adding medium VC at dl 0tIIk degree? A friend that performs cultivation under the same conditions as 111. In the results, one strain shown in Table 5 was used as a substitute for Natsucalomyces cerevisiae 1!102546 shown in Table 2, and other than adding it to α11/ll of various amino at-medium. Culture was performed under the same conditions as in Example 1. The results are shown in the $5 table 6 Table 3

Claims (1)

[Claims] t A yeast having the ability to produce 8-adenosylmethionine is cultured in a medium containing methionine/8-adenosylmethionine/
A method for producing 8-adenosylmethioni/, characterized in that the molecule Kil*I [non-sulfur amino acid that does not contain offspring, and its salt] is added to the culture medium. The genus Satucharomyces and the genus Ca/Deida belong to the genus Ha/Zenuts **-Searching range I! 1st
Method described in Section 0 5. O non-sulfur amino acids in the medium Makusha Salt (2) 11
The method described in Range Collection 1JJi for determining %IFFM where the degree is α021/lie or more.