JPS615778A - Streptomyces griseus 446-s3 - Google Patents

Abstract

PURPOSE:To produce a novel antibiotic 446-S3-1, effective mainly against streptococci and acid-fast microorganisms, having very low toxicity, and expectable to be useful as a remedy for infectious diseases of humans and animals, etc. and other applications. CONSTITUTION:A new strain Streptomyces griseus 446-S3 (FERM-P No.7416), separated from soil in YAMAGATA Prefecture, and having the ability to produce an antibiotic 446-S3-1 is cultivated under aerobic conditions to collect the aimed antibiotic 446-S3-1 from the resultant culture fluid.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to Streptomyces griseus (Streptomyces griseus).
tomyces grlseus) new strain 1I
1-53, more specifically, antibiotics≠≠6-53
This is related to Streptomyces griseus ≠ g6-53 (Feikoken Bibori No. 7t/6), which has the ability to produce -/. In the process, they isolated a new strain of Actinobacterium 4t6-33 from Yamagata soil using the usual method, and discovered that this microorganism produces a new antibiotic ≠≠6-53-/, completing the present invention. It has come to this.

The present inventors collected soil from Oaza Yamadera, Yamagata City, Yamagata Prefecture in August 1982, and smeared the water-suspended upper layer of the soil on an agar plate with a normal composition at a temperature of 1.27 to 30°C. After culturing for 3 to 5 days, the resulting microbial colonies were separated onto an agar slant with a normal composition and cultured for 1≠ days at 27 to 30°C to obtain a large number of bacterial strains. Each of these strains was cultured with shaking in a medium with a normal composition at 27 to 30°C for 3 to 5 days, and the antibacterial activity of the culture solution was tested using various test bacteria to determine which strains had antibacterial activity. Selected. As a result of more detailed testing of these strains, it was discovered that a strain named Il≠6-53 produced a new antibiotic named ≠≠6-53-7 in the culture solution. Furthermore, the purity of this strain was confirmed by performing a monospore isolation procedure.

This newly isolated microorganism has the following mycological properties, and for the reasons described below, it has been classified as a new strain of Streptomyces griseus.
It was identified as A44g-s.3.

This strain is Streptomyces sp. 446-
S 3 (Streptomyces sp, Hiroshi 44
6-53) and has been deposited with the Institute of Microbial Technology, Agency of Industrial Science and Technology, and its accession number is FERMP-7'l/6 (FERMP-7≠76).

Unless otherwise specified, the composition of the culture medium used in the following experiments is based on the identification of actinomycetes in the Patent Office's industry-specific examination standards, Applied Microbial Industry (revised 2nd edition, August 1982), pages 2g to 33. The composition of the medium used was followed.

Mycological properties of 4LIIL6-3J strain, /) Morphology 4tl, -53 strain cultured at 27°C for 2 weeks on starch/inorganic salt agar medium and yeast/malt agar medium resulted in pale green-yellow aerial mycelia. When observed under a microscope, the aerial hyphae are simply branched and gently curved.
It is also partially straight and shows tufted branches. No axle-like branches or spiral spore-bearing hyphae are observed. When observed with an electron microscope, the tip of the aerial hyphae forms a chain of 10 or more cylindrical spores, and the spores have diameters of O and S.
/, 0 μm, length σ ter/9 g μm, and the surface of the spore is smooth. Flagella, sporangia, and sclerotia are not observed.

j) Growth conditions in various media Experiments and observations were conducted by E.B. 7Yering and D. Gottlieb (Int. J. 5yst).

Bacterol, Vol. 16, p. 313, 1966). When describing colors, please refer to the 3rd edition of Nihon Shikiken Color Name Book, published by Nihon Shikiken Business Co., Ltd.
JIS symbol is shown in LA (). The following is the observation result after λ week of culture at 27°C unless otherwise specified.

(1) Growth on sucrose/nitrate agar medium is good and spreads. The attachment of aerial mycelia is poor and powdery, and is white (NZO) or very pale green-yellow (10Y).
ta//, j). The color of the back surface of the soil hyphae is pale yellow (YojY Ta/6) and bright green-yellow (10Y
1.3/litre) of soluble dye.

(2) Glucose as Daragi/Agar medium Growth is somewhat poor and flat. The attachment of aerial mycelium is also poor, and it appears as a pale green/yellow powder (lOY 7i, r), and small water droplets are sometimes observed on the aerial mycelium. The color of the back surface of the soil hyphae is a very pale green-yellow (10Y Ta//, j), and a very pale green-yellow CLOY Ta//,! j;) produces a soluble dye.

(3) Glycerin/Asuno mother goose/agar medium (Is
P-j medium) It swells and grows well. The attachment of aerial mycelia is moderately hazy and very pale reddish yellow (2,j Y Ta/
2) Very pale yellow (t, J) inside the velvet-like color.
-Y ta/ /, j) powdery parts are scattered.

The color of the back surface of the soil hyphae is grayish-yellow (YOJYza/3), and it produces a pale green-yellow CLOYli/3) soluble pigment.

(4) Starch/inorganic salt agar medium (ISP-≠ medium) Growth is good and spreads. The settlement of aerial mycelia is very good, and the central part of the colony is powdery, and the peripheral part is powdery with unevenness, and is very pale yellow-green CjGY 1. j//,!
;). The back side of the soil hyphae is bright yellow (3, JY
7/j) to produce a pale green-yellow (10Y>/j) soluble dye.

(5) Tyrosine agar medium shows very good growth and rises. Aerial mycelium is also well established, with a velpet-like part that is grayish white (Yu G, Ta 10.j) and a very pale yellow-green C! ;GY 13;//
, 3) 1. Powder-like parts with unevenness coexist. The color of the underside of the basal hyphae is yellowish brown (RiYR≠/≠), with dark yellowish brown (RiYR3/3) in places, and very pale yellow (RiYR≠/≠).
It produces a soluble dye of y/3).

(6) Growth on nutrient agar medium is poor, with a small number of white (NY, 0) aerial mycelium attached. The color of the back side of the basal hyphae is very pale yellow 1:jY
ta/3), no formation of soluble dye was observed.

(7) Yeast/malt agar medium (ISP-2 medium)
It shows extremely good growth and is slightly raised and uneven. Aerial mycelium grows very well in powder form, and its color is very pale green-yellow (10Y//J). The color of the underside of the basal hyphae is pale yellow (YojY 7/3), and it produces a bright yellow-(YojYg, 3;/ri) soluble pigment (
8) Growth on oatmeal agar medium (IsP-3 medium) is somewhat poor and flat. The attachment of aerial mycelia is also poor, and they are scattered in the form of a faint greenish-yellow (lθY ta//, 3) powder. The color of the back surface of the basal hyphae is very pale yellow (±5Y Ta/
3) to gray-yellow (yojYdo/3), and the back side of the village periphery exhibits a deep yellow <2.3-Y7//θ). Produces a soluble pigment of pale yellow (!i, 3rY 9/3).

3) Physiological properties (1) Growth temperature range Tested using starch/inorganic salt agar medium and yeast/malt agar medium at temperatures of /Jt, 20°C, 27°C, 37°C, tio°C, and <zt°C. Result, /j, C~3
It grows at each m degree of 7c, but does not grow at gu θ°C and l/Lj°C. The optimum temperature for growth is around 27°C.

(2) Liquefaction of gelatin (puncture culture was carried out at 20°C using glucose-e7''ton-zeo-latch/medium).
It is positive. In other words, no liquefaction was observed in culture 7B11./
Liquefaction begins around the pewter of the eye and progresses gradually.

(3) Hydrolysis of starch (cultivated on a starch/inorganic salt agar medium at 27°C and judged according to a conventional method) The hydrolytic power of starch is relatively strong.

(4) Coagulation and mizotonization of skim milk (Ui2 fat milk medium was cultured at 27°C and 37°C) Coagulation of skim milk was not observed at either 27°C or 37°C. At 1.27℃, bezotonization is ≠ from the white of the eyes, 37
℃, it starts from the whites of the eyes on the 10th day, progresses gradually, and almost completes at about 3M at any temperature.

(5) Production of melanin-like pigment (tyrosine agar medium,
Sect/・Yeast Iron Agar Medium, and Trigton・1−
Broth: Both were cultured at 27°C. ) No production of melanin-like pigments was observed in any of the above media.

4t) Availability of carbon sources As a result of culturing and testing at 1.27°C using Prudham-Gottlieb agar 11 medium (ISP-ta medium), D-xylose, D-glucose, D-fructose,
O-mannitol, D-Z7/'lyh-,2-hyosaricin is utilized, nuculose and inositol are of questionable utility, and arabinose, L-rhamnose, raffinose and hycellulose are of limited utility.

5) Analysis of noaminopimelic acid in the cell wall Diaminopimelic acid, which is one of the amino acids constituting the cell wall.
segawa, et al-*J, Gen, Ap
pl, Mlcrobiol, ) 2ri.

It was analyzed using the method of 31ter 3.22 (/rito3) and the fc result was LL-type.

To summarize the above results, the aerial fungus 2 with ≠4'/;-83 has simple branches, most of which are gently bent, the epiphytic part of the conidia shows tufted branches, whorled branches and spiral branches. No formation was observed, and the surface of the spores was smooth. Flagella, prespores, and sclerotia are not to be blamed. On various media, it forms basal hyphae that are very pale yellow, very pale green-yellow to yellow-brown, or dark yellow-brown. It has a very faint reddish-yellow color. In addition, it produces soluble pigments that are very bright green-yellow, very pale yellow, pale green-yellow, or dull yellow. As a result of testing at 15°C to 1.5°C, it grew between /, f°C and 77°C, and ≠
It does not grow above θ℃. It liquefies gelatin and converts skim milk into begton without solidifying it. Hydrolyzes starch well and does not produce melanin-like pigments.

D-xylose, D-glucose, D-fructose,
D-mannitol, D-utilizes lactose and salicin, sucrose and ictol are questionable, and L-7 rabinose, L-rhamnose, raffinose and cellulose are not utilized.

Noaminopimelic acid in the cell wall is in the LL-form.

From these results, this strain belongs to the genus Streptomyces,
! It clearly belongs to the Yellow Series of Lidham and Treschy's Minute L.

Therefore, a comparison was made using the ``Birshi's Manual on Determinative Bacteriology G Edition'' and the International Journal of Stematic Bacteriology, LF Sound, page 332, and as shown in Table 1, It matches well with the description of Streptomyces griseus. However, the new antibiotic
Since there was a clear difference in the production of grapes, we concluded that it is appropriate that strain ≠≠6-53 is a new strain belonging to Streptomyces griseus,
griseus) 4t11.6-33.

The microorganism according to the present invention, Streptomyces griseus
t≠6-53 has the ability to produce a new antibiotic, and its usefulness will be explained below.

Streptomyces sp. ll-411,-53 (
Antibiotics≠Hiroshi &-3 were inoculated into a nutrient-containing medium and grown aerobically.
A culture containing 3-/ is obtained. As the nutrient source, those known as nutrient sources for actinomycetes can be used. For example, commercially available carbohydrates such as starch, textrin, maltose, sucrose, crucose, glycerin, molasses, organic acids, carbon sources such as fats and oils, and defatted soybean flour, soybean flour, cottonseed flour, Nz- Amine, corn stew sol liquor,
Casein thawed product, Daysteller's Nrupuru, fishmeal, 4-zotone, meat extract, yeast extract, ammonia sulfate,
Nitrogen sources such as sodium nitrate, ammonium nitrate, and ammonia; inorganic salts such as phosphates, magnesium salts, zinc salts, potassium salts, sodium salts, and calcium salts; and trace amounts of metal salts can also be added as necessary. These substances can be used as long as they are useful for the production of the antibiotic ll-116-53-/ by the producing bacteria, and all known culture materials for actinomycetes can be used. Antibiotic≠4tg-53-
Liquid culture is preferred for efficient production of /, and the culture temperature may be within a range that allows the producing bacteria to grow and produce the antibiotic 'I'16-53-/. Culture is usually treated with antibiotics≠
This continues until ≠A-33-/ is sufficiently accumulated. For example, glycerin/f%, polybegton 0.6%, meat extract 0. liquid medium (pH 70) consisting of J%, salt 0.3%
≠≠6-53 strain cultured on agar slant medium was inoculated into
When cultured with rotary shaking aerobically at r0±/'C, culture 2
Accumulation of the target antibiotic is observed from the 8th eye, reaching the maximum between the ≠8th and 68th eyes. Antibiotic tA≠4-53-
/ Mycobacterium smegmatis was used as a test bacterium.
ls) A conventional cylindrical plate method or Eathter disk plate method using ATCC607 may be used.

In the culture obtained in this way, antibiotic l/1
．． The majority of 1A6-s3-/ is present in the culture solution or centrifuged supernatant. From those solutions antibiotics 11116-
Various methods are used to collect 33-/. That is, a method of adsorption/desorption using various adsorbents, a method of purification using various ion exchange resins, etc. can be implemented.

For example, as an adsorbent, use Diaion HP20C, a porous nonionic adsorption resin manufactured by Mitsubishi Chemical Industries, Ltd. (trade name).
When using , antibiotic ≠ 11.6-53-/ is HP,
20, and is eluted with acetone water, methanol water, etc. In addition, it is adsorbed in the hydrogen form of Amber 2ite IRC-jO (manufactured by Rohm and Haas, USA, trade name), which is a weakly acidic cation exchange resin, and is adsorbed in the hydrogen form, so that dilute acids such as 0.
Eluted with OjN hydrochloric acid. The crude powder of antibiotic 446-53-/ obtained by this method is subjected to silica column chromatography using a mixed solvent system consisting of an appropriate combination of solvents such as chloroform, methanol, benzene, ethyl acetate, and acetone. Gura.

It can be purified by method 7. In addition, weakly acidic cation exchange resins for chromatography, such as Amber 24-Shi CG-! rO (USA, ROHM & PA...-
It can also be purified using a hydrogen type (manufactured by Toyo Soda Kogyo Co., Ltd., trade name) or a resin for dull filtration a-madograph, such as Toyono or Iru HW-110 (manufactured by Toyo Soda Kogyo Co., Ltd., trade name). Also,
Repeated recrystallization using hydrous alcohol or a mixed solvent of alcohol and ethyl acetate produces highly pure antibiotics≠≠6-3
It is also possible to obtain 3-/.

Next, an example of the use of the microorganism according to the present invention will be illustrated.

Usage example / Streptomyces sp.≠IA6-53 strain (St
reptomyces Sp, II l/lly −
S 3 ) (Fiber Technology Research Institute No. 7≠76) slant culture was used as the seed, and the medium was glycerol A, 0.6% polybegtone, beef extract 0j%, and sodium chloride 0.
After adjusting the composition of 3% to p) + O, ri, 1.2
The one used was sterilized at 7°C for 75 minutes. A loopful of the inoculum was placed in a 100ml cup of the above medium and cultured with shaking at 2g°C for λ days. 2j Erlenmeyer flasks each containing the above medium were inoculated at a rate of 3 flasks, and cultured in a rotary shaker incubator (/♂θ rotations per minute) at 21~1'°C for two days.

Soow Erlenmeyer-7 Lasco,? Culture obtained by combining the cultures of J' 2. ! After adjusting t to pJ (7) with 1.2 N-* acid, it was centrifuged. The resulting supernatant was passed through a Diaion HP, 2θ resin column (20θ-) to adsorb the target substance, and washed with water. After /, 000-jO was eluted with aqueous acetone.Active category 2 obtained by elution
00 ml was concentrated under reduced pressure and further freeze-dried to obtain a brown powder. This brown powder was filled with methane.
column (manufactured by Toyo Soda Kogyo Co., Ltd.).

≠jOtd), and was developed and eluted with methanol to obtain an oily fraction (fraction number/, No. 2~/Hiroban, 20 vl/fraction). This active fraction 60- was evaporated to dryness under reduced pressure,
A brown solid was obtained. The obtained brown solid, o, soi, was dissolved in a small amount of methanol, placed on a silica gel column (Waco Pure Chemical Industries, Ltd. Wacoal C-200, ≠00d) filled with chloroform, developed with chloroform, and then Chloroborum: methanol (3:/)
The active fractions (fraction numbers 2j to 36, 10-/fraction) were obtained by developing and eluting with a mixed solvent. This active fraction L
The cd was evaporated to dryness under reduced pressure to obtain 200W of pale yellow powder. This was further placed on a silica gel column (/30d) filled with chloroform, developed with chloroform, and then sequentially using a mixture of chloroform:methanol (j:/) and chloroform:methanol (3:l). It was developed and eluted. This activity category 30. The precipitate obtained by concentrating d under reduced pressure and adding ethyl acetate was separated and dried to obtain a white powder of antibiotic 4t4t6-53-/♂0
111i+ was obtained. The physical and chemical properties of this product were as follows.

(1) Elemental analysis value c titso%, H≠%, 0 33.7
/%.

N / 2.30 chi.

(2) In molecular weight secondary ion mass spectrometry (S l -MS) analysis, (M+H)" is observed at mass number 370, (M+Na)" is observed at 392, and ('M+K)+- is observed in the box. Therefore, it is concluded that the molecular weight is j6.

(3) Melting point: /73-176℃ (4) Ultraviolet absorption spectrum: Maximum absorption in methanol (λMeG-10m(t))
ax indicates 2/g (/ls, 200), 2!7 (shoulder 7za 00). As shown in Figure 1. Also, aqueous solution,
θ, /normal hydrochloric acid, and 0- /normal hydroxide) 17
The ultraviolet absorption spectrum in an aqueous solution of aluminum does not show any characteristic absorption maximum, but only shows terminal absorption.

(5) Infrared absorption spectrum: The infrared absorption spectrum measured at B is as shown in Figure 2.The wavelength showing the absorption maximum is as follows.The unit is scratch.34t00.

2q, qO, /b♂0. /! /! ;, /, 27θl/1
0θ.

(6) Solubility in solvents: Extremely soluble in water and methanol, soluble in ethanol and acetone, slightly soluble in chloroform, and insoluble in ethyl acetate and benzene jl -Q xane.

(7) Color reaction: (@) Potassium permanganate reaction, Molisch reaction (negative) Anthrone reaction, Aniline hydrogen phthalate reaction, Ferric chloride reaction, Ninhydrin reaction (8) Basic, neutral, Acidic classification: Basic (9) Substance color: White Color Ql) Rf value Nijirikaguru Thin Layer Chroma Dog 2 Fee (TLC manufactured by Merck & Co., Ltd.)
Great, silica gel "" 254ζ thickness 0. ．． 26
The Rf value when mm) is carried out in the usual manner is -
It is a cage.

Rf value (() poroform: methanol (,!:/)
0.1A3(b)benzene: methanol (
,2:/) 0.2/(c) n-butanol:acetic acid:water(3:p/) 0.1A3(11)
Uracil and ri-sin are present in the acid hydrolyzate.

(le1+NvR(≠00MHz, D, O) δ
(ppm);/, 37C/H), /, 1. ≠(/H),
/, 7f (/+).

/, 1≠(/H), /,riJ(/H),, 2.0
(7(/H).

3.3(/H), 3.32C3H,s), 3.77(/
H,t).

1A23(/H), 1AtA(/H,t), 1AjOC
/H,t).

Hiro53(/H), lA+l-(/H), 4', noO
C/H).

1m/H, d), i';#(/H, d),! rJI(/
H, d).

Otsu, ott (/H), 771CtH, d).

04 13CNMR (/OOMHz -020)
δ(ppm):,2za:l'(t),,2f,6/(t
), 37.33(t).

Hiro 133Ct),! ;3. l0Cd>, 3 and 70(q)
.

62.72Cd), 6yo7.2(d), 7.2. ♂0(
d).

Shea, L2(d), 7g, Ri7(d), I2.39(d
).

90.71(d), 100.00Cd), 1OLIAI
Cd).

10Z7.2Cd), /! /, Lj-(d), /LZθ
g(s).

/j Ootsu, 2(s )', /67.7≠(s ), I6
6.30Cs).

I73.2θ(S), I76.4t7(s).

Usage example As a type of bacteria, 2-
was added to a test tube with a screw plug containing a pre-sterilized Hiro O tissue cloth aqueous solution 2-, mixed, and heated at -20°C.
The one that had been frozen and stored was used. Return this seed culture to room temperature and add 100 tt of culture medium per use case to j 00 gs.
L/7? Inoculate year 7 lasco and inoculate at 2fC.
The cells were cultured with shaking for days.

This seed culture solution was inoculated into 300 wt 1 volume Erlenmeyer 72 SCJO bottles containing 100-ml culture medium at a ratio of 3'16 each, and cultured with shaking at 2 g°C for 6 days.

In this case, the medium is glycerol 2θ.

One with a composition of glucose O1j, polypeptone 0.6To, beef extract 0.4J, and sodium chloride 0.3J -6, Riniiii! After heating, it was sterilized at /2/°C for 75 minutes and used.

The obtained culture 01 was centrifuged to obtain a culture supernatant.

This was applied to a Diaion HP20tMWj column (jOO
gnl) to adsorb the target substance, and after washing with water, /, S
The active fraction obtained by eluting with aqueous acetone was distilled under reduced pressure to remove acetone, and then freeze-dried to obtain 3.3 g of a brown powder. . This was dissolved in methanol and filled with chloroform.
Place it on a column (70CW) and develop with chloroform, then develop and elute in order using a mixed solvent of arophor A:///-b (s:t) and quaroform:methanol (3:l). , active fraction (fraction number 210 to 2tO).

20-/fraction) was obtained. This active fraction was concentrated under reduced pressure, and ethyl acetate was added to obtain a white precipitate, which was separated and dried under reduced pressure to obtain pale yellow powder Og. After dissolving this in water and adjusting the voice to 7.3, use a weakly acidic cation exchange resin 7 amp 2-ite ca-so.
(hydrogen type, 2sOvz) to adsorb the target substance. Next, it was washed with water and further eluted with 0.0 JN hydrochloric acid to obtain active fraction 330-. The obtained active fraction was concentrated under reduced pressure to 20%, and this was poured into Toyopearl H filled with water.
It was loaded on a column (3≠/)v) with W-≠OF, developed and eluted with water, and an active fraction j0- was obtained. This was freeze-dried and antibiotic 4146-3. ? - / white powder "3709-
I got it. The physicochemical properties of this substance were consistent with the above-mentioned physicochemical properties.

The biological properties of the above antibiotic 446-53-/ are as follows.

(υ Method established by the Antibacterial Japanese Society of Chemotherapy [Minimum Inhibitory Concentration (MI)
G) Regarding the revision of the measurement method (Kumotherapy, Kota, 7&
-72,/9za/)], the minimum inhibition of bacterial growth measured by the agar dilution method! &(MIG)
is as shown in Table 2.

In addition, Aspergillus oryzae
soryzae ) l FO! ;23ri, -1! Penlcllllum 9sA
chrysogenum ) ATCC/θθ02
, Candida albicans (Candld6 al
blcans) 3/lI-7 and Saccharomyces cerevisiae (Saccharomycescere)
The minimum inhibitory concentration for fungi such as Vlslae ) l F OO, 20j is t 00 m
cg/- or more.

(2) When a physiological saline solution is administered intravenously with toxicity, /
,000 rl#g/to! No. 9 showed no toxicity at all.

As described above, the antibiotic ≠≠A-33-/ is mainly effective against streptococci and acid-fast bacteria, and has extremely low toxicity, so it is expected to be used as a therapeutic agent for infectious diseases in humans, animals, etc., and for other uses.

As a result of comparing the above-mentioned physicochemical properties and biological properties with those of known antibiotics, no corresponding substance was found, so antibiotic G≠6-53-/ was determined to be a new antibiotic.

Furthermore, by 1H-NMR, 13C-NMR and amino acid analysis of the acid hydrolyzate, antibiotics≠u6S3
It was confirmed that it is a so-called nucleoside antibiotic containing a -/GA t 5 syl group and a residual group of lysine units. On the other hand, among the known nucleoside antibiotics, there are no substances containing lysine, so lI
If t≠6-93-/, it can be determined that it is a new antibiotic. ,

[Brief explanation of the drawing]

Figure 1 shows the ultraviolet absorption spectrum (measured in g in methanol) of the antibiotic ≠≠A-83-/, and Figure 2 shows the infrared absorption spectrum (CKBr method). (Figure 1 wavelength c plane) t/evening ig Hand Vt correction book Showa 1988 23rd day 1, Incident indication Patent application No. 12676 of 1982
No. 5, No. 3, Relationship with the case of the person making the amendment Applicant Name Kanto Ishi Pharmaceutical Co., Ltd. 4, Agent 6, Subject of amendment Column for detailed explanation of the invention in the specification //P-, (11 Specification On page 20, lines 11-15, “Also...
...It is only shown. (Delete “basic” in line 13 of page 21 of the same book as “weakly acidic.”)
Correct.

Claims (1)

[Claims] Streptomyces griseus 446-S3 with the ability to produce antibiotic 446-S3-1
No. 416.