JPS63215689A - Lampteroflavin and production thereof - Google Patents

Abstract

NEW MATERIAL:A compound, expressed by the formula and having light emitting property. USE:A light emitting substance useful as a labeling and tracing substance in the medical field. PREPARATION:A fruit body of Lampteromyces japonicus Sing. (preferably fold part) is treated with water while being aerated and the resultant extract solution is subjected to fractionation treatment to afford the compound expressed by the formula. Furthermore, the fractionation treatment is preferably carried out by octadecylsilanol (ODS) column chromatography.

Description

Detailed Description of the Invention (Industrial Application Field) The present invention relates to a physiologically active compound represented by the following two-dimensional formula (prior art) Riboflavin is known as vitamin B2, and is an important compound involved in various chemical reactions in living organisms. It is a substance.

Laapteromyces japonl
cus) is known as a poisonous mushroom, and it has been reported that fruiting bodies grow from decaying leaves around October every year, and that when the cap opens, the folds (spores, etc.) of the fruiting bodies emit light.

In recent years, Iaa + pterol has been added to the back as an anti-cancer substance.
, II Iudln, 5esqutterupen,
have been extracted and reported. (1963) Similar components were also found in the American jack mushroom, and in 1970, 1
lludins and ergotatetraene were thought to be involved in the photic bioluminescence phenomenon.

A small number of compounds of bioluminescent substances such as fireflies are known, and they have already been industrialized as reagents for quantifying calcium and ATP.

(Problems to be Solved by the Invention) Luminescent substances can be used in various ways depending on their emission wavelength and intensity of light energy. For example, some wavelengths of light are used for their strong cell-killing and bactericidal properties, while others are used for the purpose of absorbing harmful rays.
It was predicted that they would be used as measurement reagents, and the discovery of various luminescent substances was desired.

Known luminescent substances of biological origin, such as luciferin, have different emission wavelengths and mechanisms from the substances of the present invention, but they are relatively expensive and have defects that limit their purpose of use. .

Moonlit night mushroom (Lampteromyces japonfc)
us) is known as a poisonous mushroom, and the presence of anticancer substances has been reported in recent years, but the fruiting bodies can only be harvested a few times a year, and the luminescence comes from the mushroom folds, making it difficult to secure raw materials. However, industrial utilization has not yet been investigated.

(Means and effects for solving the problem) The present inventor:
Moonlit mushroom (Lampteromyces japonlc)
Regarding the industrial use of uS, we conducted research to elucidate the luminescent phenomenon, established an industrial production method for the luminescent raw material, and recognized the existence of a luminescent substance, which we named lampteloflavin.

The lamp teroflavin according to the present invention has a maximum wavelength of 524n.
It has a luminescent property of m and has the property of being decomposed into riboflavin and sugar by enzymes such as ribosidase.

(Effects of the Invention) The compound is a ribosyl derivative of riboflavin, and has a physiologically active action as a riboflavin precursor (derivative), and can also be used as a luminescent reagent for diagnosing diseases and tracking the progress of symptoms after treatment. It is used as a harmful ray absorber.

The present invention will be explained in more detail with reference to Examples below.

Examples 1 to 4 Separation and production of Tsukiyo Mushroom fruiting body folds 3. 12.5 L of distilled water was added to 7 kg, and the components were extracted for 19 hours under mechanical oxygen flow conditions at pH 4 and 16 degrees Celsius. The extract was filtered through Celite, and 9.6 L of the filtrate was filtered through Develosll.
(Nomura Chemical) ODS (octadecylsilanol)
-W (100-200) Stainless steel column (20X
800a+a+), and then the column was washed with 3 L of distilled water and eluted with 4 L of methanol. The residual oil obtained by concentrating the methanol eluate under reduced pressure was further purified by Dovelosll.
Lop OD S glass column (28X 310+u
) to adsorb the residual oil on the column, and add 300 mL of distilled water.
After washing with water, the target component is eluted with methanol at a flow rate of 6IIIL/min. The same component exhibits macroscopic fluorescence under 365n1M ultraviolet light.

(Rampteloflavin crude yield: 1.4 g) The eluate was concentrated under reduced pressure, and the residual oil was collected in Develosll 0D.
Equivalent elution method (2
5% methanol/water; 6 mL/min) to obtain lampteloflavin. (Yield: 0.8 g) The following results were obtained due to the difference in purification conditions.

In Example 2, 3 kg of mushroom folds in a cell-dead state were added, and in Example 4, the pH was adjusted to 3 with hydrochloric acid.

Example 5 Isolation and production of the substance of the present invention from mycelium Spores of Tsukiyo mushroom were suspended in sterile water and achromycin (0,0
15%) containing potatoes, sucrose at 23 degrees Celsius
Culture for ~2 weeks. (Contains 20% potato, 2% sucrose, and 2% agar in distilled water) Collect the separated colonies,
The mixture was poured into a Sakaguchi flask containing 180 ml of potato and sucrose solution, and cultured with shaking at 23°C and 120 rpm.

In order to sequentially observe the productivity of luminescent substances, the luminescence intensity of the growing mycelium was measured by the amount of oxygen passing through the photometer AF+ tube containing the 12 solution in which the mycelia had been washed.
Light intensity was monitored with a photometer.

The maximum luminescence of the mycelium is observed at the mouth for two weeks after culturing. When performing normal tank culture, for two weeks after culture, 41 microorganisms
Cultivation is terminated at around 1000 mg/100 ml, and lampteloflavin can be obtained by ODS chromatography treatment in the same manner as in Examples 1 to 4.

The substance of the present invention has the following physicochemical properties.

I゛M construction type %formula%

[Brief explanation of the drawing]

Figure 1 shows the ultraviolet absorption spectrum of lampteloflavin. Figure 2 shows the fluorescence spectrum of lampteloflavin. Tekinaga (nm) Wotenkoon (nml procedure correction writing method) June 17, 1986

Claims (5)

[Claims] (1) Lampteroflavin is shown below in the planar form. ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ (2) Moonlit mushroom (Lampteromyces japo)
A method for producing a luminescent substance lampteloflavin, which comprises collecting the fruiting body of L. nicus, cultivating it artificially or culturing cells, and subjecting it to fractionation treatment. (3) Moonlit mushroom (Lampteromyces japo)
3. The method for producing the luminescent substance lampteloflavin according to claim 2, characterized in that the fruiting body folds of P. nicus are used. (4) Moonlit mushroom (Lampteromyces japo)
A method for producing a luminescent substance lampteloflavin, which comprises culturing and fractionating the mycelium of L. nicus. (5) A method for producing the luminescent substance lampteloflavin according to claims 2 to 4, wherein the fractionation treatment is performed using ODS (octadecylsilanol) column chromatography.