JPH0292291A - Si-4155ap3 substance, its production and use thereof - Google Patents

Abstract

PURPOSE:To obtain SI-4155AP3 substance useful as a plant growth regulator, a herbicide and an antibacterial agent by culturing a specific microbial strain belonging to genus Streptomyces. CONSTITUTION:The objective SI-4155AP3 substance has a structural formula expressed by the formula and is produced by culturing a microbial strain belonging to genus Streptomyces and capable of producing SI-4155AP3 substance, extracting the cultured product and purifying the extract. The microorganism is e.g., Streptomyces SP4155 (FERM P-10236). The culture of the microorganism is carried out preferably under aerobic condition at 23-33 deg.C for 3-10 days.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a novel SI-41558P3 substance, its production method and uses. SI-4155AP3 substance is a physiologically active substance produced by microorganisms, and is useful as a plant growth regulator, herbicide, and antibacterial agent.

[Prior Art 1 Problems to be Solved by the Invention] Various physiologically active substances produced by microorganisms are known, but
The SI-4155AP3 substance of the present invention is a new substance,
Its manufacturing method is also unknown.

[Means for Solving the Problems] The present invention provides SI-4155A having the following physical and chemical properties.
Regarding P3 substances.

(a) Appearance: white powder (b) Elemental analysis values: carbon 65.35%, hydrogen 9.13%
.

Nitrogen 0% (8) Molecular weight 740 (from FAB US) (d) Melting point: 108.0 -110°C (e) Specific optical rotation = [α] 38-2o, o.

(c = 0.014, methanol) (f) Infrared absorption spectrum: shown in Figure 1 (measured in Br tablet) (g) Ultraviolet absorption spectrum, shown in Figure 2 (measured in methanol) (h) 11(-Nuclear 6i1 gas resonance spectrum: Shown in Figure 3 (CDC1') (Su) 13c-Nuclear magnetic resonance spectrum: Shown in Figure 4 (CDCJ3) (N) Solubility: benzene, ethyl acetate, chloroform,
Soluble in acetone and methanol, insoluble in water and n-hexane Color reaction: Positive for potassium permanganate and iodine Furthermore, the present invention relates to the SI-4155AP3 substance, which belongs to the genus Streptomyces and has the above-mentioned physicochemical properties. SI-415 is grown from the culture by culturing microorganisms that have the ability to produce SI-415.
SI-415 characterized by collecting 5AP3 substances
5AP3 substance manufacturing method and the SI-4155AP3
The present invention relates to plant growth regulators, herbicides, and antibacterial agents containing the substance as an active ingredient.

The SI-4155AP3 substance, which is a new substance of the present invention, is
A microorganism that belongs to the genus Streptomyces and has the ability to produce 5i4155AP3 substance is cultured, and the culture is extracted with SI-
Obtained by collecting 4155AP3 substance.

As a microorganism that has the ability to produce 51-41558 P3 substances, Streptomyces sp., which was newly discovered by the present inventors, is used.
4155 strain, and the microorganism has the following mycological properties.

■, Morphology Presence of aerial hyphae Substantial hypha division Presence of sporangia Spore motile Chain of spores Branching of sporophyte Whorled spore Chain morphology Pseudospore Cysticionia fungi Nucleotidial sporangium club Fusion of sporangial hyphae + + Straight and wavy (RF) + (observed on yeast malt agar medium, ISP'-2) ◆ International Streptomyces Project Selected media Growth status on various media Ill , Physiological properties Growth temperature range: 15-38°C (optimum temperature range 28-34°C) Growth pi (range; 5, θ-1O10 (optimum pH range 5.5-8.5) Nocturnalization of gelatin: Starch Hydrolysis of: + Hydrolysis of casein: + Coagulation of milk: Production of melanin pigment (4th day) Tryptone/yeast broth Peptone/yeast Iron agar medium Tyrosine agar medium (100th day melanin pigment production) Cell wall type: Type I Main quinone molecular species: M-9 (I(a), MK-9
(lla).

-9()+4) to M IV, assimilability of carbon sources D-xylose: L-arabinose: + D-glucose: + D-fructose: + Sucrose: + Raffinose: ± L-rhamnose: + Inositol: + D-’f nitol: ± cellulose (-: Not to be used, ±: I doubt whether to use it.

+: Utilize (slightly inferior to D-glucose), +
: Indicates that it is strongly utilized (to the same extent as D-glucose). ) Since the SI-4155 strain, which has the above-mentioned characteristics, is considered to be a strain belonging to the genus Streptomyces, the present inventors identified the wood fungus as Streptomyces sp.
Treptomyces sp.) 4155 strain. The wood fungus is FERed to the Institute of Microbial Technology, Agency of Industrial Science and Technology.
It has been deposited as MP-10236. In addition, in the present invention, even mutant strains obtained by artificially mutating or naturally mutating wood fungi can be used as SI-4155.
Anything capable of producing AP3 substances can be used.

As a medium for culturing the above-mentioned microorganisms, one containing a carbon source, a nitrogen source, etc. and in which the microorganisms can grow is used. Various carbon sources can be used, including glucose, sucrose, fructose, rhamnose, and arabinose. In addition, nitrogen sources include ammonium nitrate, ammonium sulfate, meat extract,
Corn steep liquor, peptone, dry yeast.

Mention may be made of inorganic or organic nitrogen compounds such as soy flour. In addition, inorganic salts such as common salt, phosphates, calcium, zinc, magnesium, and iron, and nutritional substances necessary for the growth of microorganisms can be added to the medium as necessary.

The above microorganisms are cultured under aerobic conditions at a temperature of 20 to 35°C.
, preferably at 23 to 33°C for 3 to 10 days, preferably 4 to 6 days, and at a pH of 5 to 9, preferably 6 to 8, whereby SI-41558P3 substance is produced in the culture. Accumulated. The culture is stopped when the production amount of the substance in the culture reaches the maximum, and 5 t is removed from the culture.
-4155AP3 material is collected and purified.

For purification, for example, a method may be used in which an organic solvent such as ethyl acetate is added to the culture to perform extraction, the extract is concentrated, and then separated using silica gel column chromatography.

5i4155A, a new substance obtained by the above method
Since P3 substance has plant germination inhibitory activity, it is classified as SI.
In addition to being used as a plant growth regulator containing SI-41558P3 as an active ingredient, it can also be used as a herbicide and as an antibacterial agent using its antibacterial activity. can.

When used as a plant growth regulator, S
Hydrating agents and flowable agents containing I-4155AP3 substances,
Plant growth can be regulated by making it into a formulation such as an emulsion or powder and applying it to soil or a cultivation solution for hydroponics in a dosage range that does not kill the target plant.

When used as a herbicide, SI-41 is used as an active ingredient.
Problematic weeds can be killed by making preparations such as permanent preparations, flowable preparations, emulsions, powder tablets, etc. containing the 558P3 substance and applying them to the soil from before the weeds germinate to early after they germinate.

In addition, when used as an antibacterial agent, target harmful microorganisms can be controlled by making the formulation into a formulation according to the respective use, such as medical or industrial use.

[Example] Next, the present invention will be explained by examples.

Example 1 Bridham Gottlieb Pharmamedia medium 125 as shown in Table 1 in a 500 mj' flat bottom shake flask.
After adding m1 and sterilizing it in an autoclave, the results are shown in Table 2. Streptomyces sp. 4155 (FERM P-10236) was inoculated from the slant of a modified Bennett medium and cultured at 30° C. for 24 days.

Table 1 glucose Huamamesoia to 82PO4 2HPO4・3H20 MgSO4・7H20 Trace salt solution.

Distilled water 0g 0g 2.38g 5, [i5g 1g 1mJ+ 000mj pH7.0~7.4 ◆Trace salt solution CuSO4・5H200,64g FeSOn・7) 120 0.11 gMn
CI12”4Hx0 0.78 gZnSO
4.7) 1zOO, 15g Distilled water 100mj 1 Main 1 Glucose 10g Polypeptone
2g Meat extract 1g Yeast extract 1g Distilled water 10100O! After completion of the culture, the culture solution obtained by centrifugation was adjusted to neutrality and extracted with ethyl acetate. Next, the extract was dehydrated by adding anhydrous sodium sulfate, and then concentrated to dryness to obtain an oily crude extract.

The obtained crude extract was subjected to silica gel column chromatography (n
- Elution was performed using ethyl acetate (increasing the concentration of ethyl acetate by 20% from 100% hexane), and n-hexane: ethyl acetate =
Fractions from 20:80 to 0:too were collected. Next, both obtained fractions were purified again by silica gel column chromatography using the solvent system n-hexane: ethyl acetate (30 nia 0), and further purified by silica gel column chromatography using the solvent system diethyl ether; ethanol (
Silica gel column chromatography was performed twice using 98;2) to obtain 0.8 mg of SI-4155AP3 substance as a white powder.

Example 2 Filter paper (Whatman No.,
2) and SI-4155AP3 obtained in Example 1.
The substance was dissolved in acetone and added to a petri dish to a predetermined concentration. After air-drying, add 3 mj of ion-exchanged water and add pre-germinated lettuce seedlings (Great L).
Sow 7 grains of ake366) and put it in an artificial weather machine (27℃, approx.
The lettuce seedlings were grown for 3 days in 000 lux) and the root length of the lettuce seedlings was measured. The measured value was the average value excluding the longest and shortest values. From the measured values, the root elongation rate of the test plot was calculated, assuming that the length of the roots of lettuce seedlings fed only ion-exchanged water was taken as 100. The results are shown in FIG.

As is clear from FIG. 5, the SI-4155AP3 substance has the activity of inhibiting plant growth.

Example 3 After culturing the test bacteria shown in Table 3 in a bouillon liquid medium at 30°C for 24 hours, the culture solution was placed on a bouillon agar medium at a concentration of about 1%.
A test plate was prepared by adding B to a Petri dish. Next, SI-4155^P obtained in Example 1
A paper disk was immersed in a solution of the three substances dissolved in methanol at a concentration of 111g/ff1R, and after air-drying, the vapor disk was placed on the assay plate and incubated at 30°C for 24 hours. After completion of the culture, antibacterial activity was evaluated based on the presence or absence of inhibition date. The results are shown in Table 3. In addition, + in the table indicates antibacterial activity, and - indicates no antibacterial activity.

Table 3 [Effects of the Invention] The SI-4155AP3 substance of the present invention is a physiologically active substance produced by microorganisms, and is useful as a plant growth regulator, herbicide, and antibacterial agent.

[Brief explanation of drawings]

FIG. 1 is an infrared absorption spectrum of the SI-4155AP3 material. FIG. 2 is an ultraviolet absorption spectrum of the SI-4155AP3 material. Figure 3 shows SI-415
It is '11-nuclear magnetic resonance spectrum of 5AP3 substance. FIG. 4 is a 13C-nuclear magnetic resonance spectrum of SI-4155AP3 material. FIG. 5 shows the root elongation rate of the test plots in Example 2. Patent Applicant: Hiroshi 1) Yang No. 5 Figure SI-IH55AP3 ↑Double Sleeping Concentrated L (ppm) Procedural Amendment (Voluntary) September 1999

Claims (5)

[Claims] (1) SI-4155AP with the following physical and chemical properties
3 substances. (a) Appearance: white powder (b) Elemental analysis: carbon 65.35%, hydrogen 9.13%
, nitrogen 0% (c) Molecular weight: 740 (from FABMS) (d) Melting point: 108.0-110°C (e) Specific optical rotation: [α]_D^2^■-20.0° (c
=0.014, methanol) (f) Infrared absorption spectrum: shown in Figure 1 (KB_r
(Measured in tablets) (G) Ultraviolet absorption spectrum: Shown in Figure 2 (Measured in methanol) (H) ^1H-Nuclear magnetic resonance spectrum: Shown in Figure 3 (
CDCl_3) (li)^1^3C-Nuclear magnetic resonance spectrum: Shown in Figure 4 (CDCl_3) (n) Solubility: benzene, ethyl acetate, chloroform,
Soluble in acetone and methanol, insoluble in water and n-hexane (Le) Color reaction: Positive for potassium permanganate and iodine (2) S according to claim 1, which belongs to the genus Streptomyces
A method for producing SI-4155AP3 substance, which comprises culturing a microorganism capable of producing I-4155AP3 substance and collecting SI-4155AP3 substance from the culture. (3) A plant growth regulator containing the SI-4155AP3 substance according to claim 1 as an active ingredient. (4) A herbicide containing the SI-4155AP3 substance according to claim 1 as an active ingredient. (5) An antibacterial agent containing the SI-4155AP3 substance according to claim 1 as an active ingredient.