(1 9) Japan Patent Office (JP)
`
`(1 1)Japanese Unexamined Patent Publication
`No. 853-1 19872
`
`(51) Int. Cl. 2
`
`ID Code
`
`C 07C 103/52
`A61 K37/02
`
`107
`ADZ
`
`Japanese Unexamined Patent Publication
`(52) Japanese Classification
`JPO File No.
`
`16 E 362
`30G 196
`30 H 612
`
`7242-44
`6617-44
`5727-44
`
`(43) Japanese Unexamined Patent Publication
`Date: October 19, 1978
`
`(54) Trtte ofthe Invention: New peptides and antimicrobial agents
`
`(21 ) PatentAppIication No.: 852-355
`
`(2) Filing Date: March 29, 1977
`
`Requestfor Examination: Not yet made
`Numberof inventions: 4
`
`Total @es: 6
`02) Inventor: Shunpei Sakakihara, 2-23-3 Fujishirodai,
`
`Suita City
`
`02) Inventor: Yoshihiro Nlasui, 2-1-6 Chokoji-Ifita,
`
`(72) Inventor: Hiroshi Nta, 1616, 2-5 Higashi-lzumigaola,
`
`Toyonaka City
`
`Toyonaka City
`
`01 )Applicant: Suntory IJmited, 2-1-1 Dojima-I-Iarr‘adori,
`
`(72) Inventor: Ryuji Tanaka, Tornita Danchi 2-502,
`
`IGta-ku, Osaka City
`
`1319 IVIakiIa-cho, Takatsuki City
`
`U4)Agent:: Toshb Takigawa, Patent Attorney
`
`Specification
`
`(4) Antimicrobial agents containing a peptide having 13 amino
`
`1.TrtIe of the Invention
`
`acids in orderoftryptophan, histidine, tryptophan, Ieucine,
`
`New peptides and antimicrobial agents
`
`glutamine, D-Ieucine, lysine, proline, glufdne, glutamine, proline,
`
`2. Claims
`
`methionine and tyrosine.
`
`(1 )A peptide having 12 amino acids in orderof histidine,
`
`3. Detailed Explanation ofthe Invention
`
`tryptophan, Ieucine, glutamine, D-Ieudne, lysine, proline, glycine,
`
`The present invention relates to new peptides and antimicrobial
`
`glutamine, proline, methionine and tyrosine.
`
`agents containing these peptides. Awide range of peptides have
`
`(2)A peptide having 13 amino acids in order oftryptophan,
`
`conventionally been isolated or synthesized, and it has been
`
`histidine, tryptophan, Ieucine, glutamine, D-Ieucine, lysine, proline,
`
`known that some ofthem have pewliar adion including
`
`glufcine, glutamine, proline, methionine and tyrosine.
`
`antimicrobial adion. Nevertheless, we do not knowany peptides
`
`(3)Antimicrobial agents containing a peptide having 12 amino
`
`that show satisfactory antimicrobial adion yet. The present
`
`acids in orderof histidine, tryptophan, Ieudne, glutamine, D-
`
`inventors succeeded in isolating peptides from media for
`
`Ieucine, lysine, proline, glycine, glutamine, proline, methionine
`
`culturing bacteria and found that some peptides in the course of
`
`and tyrosine.
`
`synthesis had superior antimicrobial adion as well.
`
`-513-
`
`

`

`Japanese Unexamined Patent Publication No. 853-1 19872 (2)
`
`Present invention relates to a peptide having 12 amino acids in
`
`the peptides according to the present invention asfaras the
`
`orderof histidine, tryptophan, leucine, glutamine, D-leudne, lysine,
`
`aforementioned amino acid sequences are kept Abbreviations
`
`proline, glycine, glutamine, proline, methionine and tyrosine
`
`(hereinafter referred toas “peptide A”), a peptide having 13
`
`used in the explanation ofthe synthetic method are first given
`below
`
`amino acids in orderoftryptophan, histidine, tryptophan, leudne,
`
`Boc: t-butybxycarbonyl, le: benzyl, Tyr: tyrosine, Met:
`
`glutamine, D-leucine, lysine, proline, glufcine, glutamine, proline,
`
`methionine, Pro: proline, Gln: glutamine, Gly: glycine, Lys: lysine,
`
`methionine and tyrosine (hereinafter referred to as “peptide B”)
`
`Leu: leudne, His: histidine,Trp: tryptophan, HOBT:
`
`and antimicrobial agents containing these peptides.
`
`hydroxybenzotriazole, WSCI: N-ethyl- N,N’-
`
`The peptides according tothe present invention show
`
`dimethylaminopropyl carbodiimide, DCHA dicycbhexyl
`
`antimicrobial adion against a variety offungi and bacteria,
`
`ammonium salt, ONSu: N-succiimide ester, ONp: p-nitrophenyl
`
`especially against Pseudomonas pyocyaneum and plant
`
`ester, ZoCl: ortho chbro benzybxycarbonyl, Tos: para toluene
`
`pathogens, and, therefore, can be used forvarbus ways
`
`sulfonyl, THF: tetrahydrofurane, DMF: dimethylforrnamide,
`
`including agricultural chemicals, fungicidal paints and therapeutic
`
`DMS: dimethyl sulfide, TFAC: trifluoroacetic acid, and TEF:
`
`agents for athlete’s foot
`
`triethylamine.
`
`Adescription ofone synthetic method ofthe present invention
`
`Synthesis of peptide A
`
`is given bebw. However, any synthetic method may be used for
`
`Synthesis of Boc-D-Leu-Lys (ZoCI) —Pro-OBz|:
`
`Dissolve oily Boc-Lys (ZoC|)—Pro-OBz| (9 g, 15 mmol) prepared
`
`Yield: 8.5 g (90.7%).
`
`bya normal method using Boc-Lys (ZoC|)—OH and Pro-Ole in
`
`Synthesis of Boc-GIn-D-Leu-Lys (ZoC|)—Pro-OH:
`
`TFAC (30 ml) under cooling. Stir the solution for 10 minutes
`
`Dissolve oily Boc-D-Leu-Lys (ZoC|)—Pro-OH (8.5 g, 13.6
`
`under cooling and then for40 minutes at room temperature.
`
`mmol) in TFAC (30 ml) under cooling. Afterstirring for 10
`
`Remove excessive TFAC under reduced pressure. Dissolve
`
`minutes under cooling and for45 minutes at room temperature,
`
`residue in DMF (30 ml), and neutralize itwith TEA under cooling.
`
`remove excessive TFAC under reduced pressure. Add etherto
`
`Add Boc-D-Leu-ONSu (4.9 g, 15 mmol), and stirthe solution for
`
`residue to make it powder. Dry itfor2 hours over NaOH under
`
`one hour under cooling and for 90 hours at room temperature.
`
`reduced pressure.
`
`Dissolve the readion liquid in ethyl acetate (300 ml). Wash the
`solution with 1N HCI, water, 5% sodium bicarbonate water and
`
`Dissolve the powder in DMF (24 ml), and neutralize itwith TEA
`
`under cooling. Add Boc-Gln-ONp (6 g, 16.3 mmol), and stirfor
`
`water. Dry it oversodium sulfate. Remove ethyl acetate under
`
`one hour under cooling and for 65 hours at room temperature.
`
`reduced pressure to produce specified oily substance.
`
`Then, add N, N’-dimethy|propane diamine (1 .5 ml), and stirfor2
`
`Yield: 10.5 g (98%).
`
`hours. Adjust pH to 1 with 1N HCI. Extractwith ethyl acetate (200
`
`Synthesis of Boc-D-Leu-Lys (ZoC|)—Pro-OH:
`
`ml), wash with water, and dry over sodium sulfate. Remove ethyl
`
`Dissolve oily Boc-D-Leu-Lys (ZoC|)—Pro-OBz| (10.5 g, 14.7
`
`acetate under reduced pressure. Add etherto residue to make it
`
`mmol) in methanol (10 ml) and dbxane (25 ml). Add 1N NaOH
`
`powder. Dry the powder obtained by reprecipitating with ethyl
`
`(14.7 ml) under cooling. Stirthe solution for2 hours. After
`
`acetate / ether over phosphorus pentoxide for24 hours.
`
`adjusting pH to 7 with 1N HCI, remove methanol and dbxane
`
`Yield: 6.4 g (63%).
`
`under reduced pressure. Add 5% sodium bicarbonate water,
`
`and wash itwith ether. Adjust pH of the aqueous layerto 2with
`
`Elemental analysis: as 035H58010N60| - H20
`Calculated values: C, 54.50%; H, 7.19%; N, 10.90%.
`
`1N HCI under cooling. Extractwith ethyl acetate (200 ml). After
`
`Analytical values: C, 54.48%; H, 7.17%; N, 10.65%.
`
`washing with water, dry itover sodium sulfate. Remove ethyl
`
`acetate under reduced pressure to produce specified oily
`substance.
`
`-514-
`
`

`

`Japanese Unexamined Patent Publication No. 853-1 19872 (3)
`
`Synthesis of Boc-Leu-Gln-D-Leu-Lys (ZoCl)- Pro-OH:
`
`Analytical values: C, 55.04%; H, 7.41%; N, 11.01%.
`
`Dissolve Boc-Gln-D-Leu-Lys (ZoCl)—Pro-OH (5.3 g, 7 mmol)
`
`Synthesis of Boc-Gly-Gln-Pro-Met—Tyr (2.6-Clz-le)—Ole:
`
`in TFAC (20 ml) under cooling. Alterstirring for 10 minutes under
`
`React Boc-Tyr(2.6-Clz-le)with benzyl bromide bya known
`
`cooling and for40 minutes at room temperature, remove
`
`method to produce Boc-Tyr(2.6-Clz-le)—Ole. Then, read it
`
`excessive TFAC under reduced pressure. Add etherto residue
`
`with Boc-lVlet (prepared from Boc-lVlet-DCHA) bya normal
`
`to make it powder, and dryfor2 hours over NaOH under
`
`method to produce Boc-lVlet— Tyr (2.6-Clz-le)—Ole. Read it
`
`reduced pressure.
`
`with Boc-Pro-ONSu, Boc-Gln-ONp, and then Boc-Gly-ONSu by
`
`Dissolve the powder in DMF 0 ml), and neutralize itvvith TEA
`
`a known method to produce Boc-Gly-Gln-Pro-lVlet—Tyr (2.6-Clz-
`
`under cooling. Add Boc-Leu-ONSu (2.8 g, 8.4 mmol), and stirfor
`
`le)—Ole.
`
`one hour under cooling and for 90 hours at room temperature.
`
`Synthesis of Boc-Leu-Gln-D-Leu-Lys (ZoCl)- Pro- Gly-Gln-Pro—
`
`Pourthe readion liquid into ice cold water (1 00 ml), and extract
`
`Met-Tyr (2, 6-Clz-le)—Ole:
`
`with ethyl acetate (200 ml). Afterwashing with 1N HCl and then
`
`Add DMS (1 ml) to Boc-Gly-Gln-Pro-lVlet—Tyr (2.6-Clz-le) —
`
`with water, dry oversodium sulfate. Remove ethyl acetate under
`
`Ole (2.4 g, 2.5 mmol). Dissolve the solution in TFAC (10 ml)
`
`reduced pressure, and make powdervvith methanol/ ether. Dry
`
`under cooling, and stirfor 10 minutes under cooling and for 35
`
`the powder obtained by reprecipitating with methanol / ether over
`
`minutes at room temperature. Remove excessive TFAC under
`
`phosphorus pentoxidefor24 hours under reduced pressure.
`
`reduced pressure. Add 7.1N HCl/dioxane (0.42 ml, 3 mmol)to
`
`Yield: 4.5 g 02%).
`
`residue under cooling. Then, add etherto make powder, and dry
`
`Elemental analysis: as C41H64011N7C| - 1.5HZO
`Calculated values: C, 55.11%; H, 7.59%; N, 10.97%.
`
`itfor2 hours over NaOH under reduced pressure.
`
`Dissolve this powder in DMF (10 ml) and THF (20 ml) together
`
`the solution in TFAC (20 ml) under cooling, and stirfor 10
`
`with Boc-Leu-Gln-D-Leu-Lys (ZoCl)- Pro-OH (2.35 g, 2.63
`
`minutes under cooling and for35 minutes at room temperature.
`
`mmol) produced above and HOBT (340 mg, 2.5 mmol). Add
`
`Remove excessive TFAC under reduced pressure. Add ether
`
`WSCI (0.46 ml, 2.5 mmol) dropwise under cooling, and stirfor
`
`to residue to make it powder, and dry itover NaOH for2 hours
`
`one hour under cooling and for 15 hours at room temperature.
`
`under reduced pressure.
`
`Dissolve it in ethyl acetate (500 ml) after removing THF under
`
`Dissolve the powder in DMF (12 ml), neutralize with TEA under
`
`reduced pressure. Washwith 1N HCl, water, 5% sodium
`
`cooling. Add Boc-Trp-ONSu (830 mg, 2.07 mmol), and stirfor
`
`bicarbonate water, and then water. Alter removing ethyl acetate
`
`one hour under cooling and for 88 hours at room temperature.
`
`under reduced pressure, add etherto residue to make it powder.
`
`Pourthe readion liquid into ice cold water (300 ml)to make it
`
`Reprecipitate ittwo times with methanol / ether, and dry the
`
`powder. Filter it, and dry the powder obtained by reprecipitating
`
`powder over phosphorus pentoxide for24 hours.
`
`with ethyl acetate / ether over phosphorus pentoxide for 24 hours
`
`Yield: 3.7 g (85%).
`
`Elemental analysis: as C81H11OO18N188CE - 2H20
`Calculated values: C, 56.29%; H, 6.65%; N, 10.5%.
`
`Analytical values: C, 5626%; H, 6.47%; N, 10.42%.
`
`under reduced pressure.
`
`Yield: 2.9 g (88%).
`
`Elemental analysis: as C92H1ZOO18N15SCE - 5HZO
`Calculated values: C, 5.44%; H, 6.55%; N, 10.92%.
`
`Synthesis of Boc-Trp-Leu-Gln-D-Leu-Lys (ZoC|)- Pro- Gly-Gln-
`
`Analytical values: C, 5.29%; H, 6.41%; N, 10.78%.
`
`Pro-Met—Tyr (2, 6-Clz-le)—Ole:
`
`Add DMS (2 ml)to Boc-Leu-Gln-D-Leu-Lys (ZoC|)- Pro- Gly-
`
`Gln-Pro-lVlet—Tyr(2, 6-ClZ-le)—Ole (3 g, 1.73 mmol). Dissolve
`
`-515-
`
`

`

`Japanese Unexamined Patent Publication No. 853-1 19872 (4)
`
`Synthesis of Boc-l-lis-TrpLeu-Gln-DLeuLys (ZoCl)- Pro- Gly-
`
`was eliminated, ard therefore the productwas a mixture of Boc-
`
`Gln-Pro-lVlet—Tyr(2, 6-Clz-le)—Ole:
`
`His-Trp-Leu-Gln-D-Leu-Lys (ZoCl)- Pro- Gly-Gln-Pro-lVlet—Tyr (2,
`
`Add DMS (1 ml) to Boc-Trp-Leu-Gln-D-Leu-Lys (ZoCl)- Pro-
`
`6-Clz-le)—Ole ard Boo-His (Tcs)-Trp-Leu-Gln-D-Leu-Lys
`
`Gly-Gln-Pro-lVlet—Tyr(2, 6-Cl2-le)—Ole (2.1 g, 1.1 mmol).
`
`(ZoCl)- Pro- Gly-Gln-Pro-lVlet—Tyr (2, 6-Clz-le)—Ole. Neither
`
`Dissolve the solution in TFAC (15 ml) urderccoling, ard stirfor
`
`physical constants noryields are described here. In the following
`
`10 minutes urderccoling ard for35 minutes at room
`
`reaction, the T06 radical was completely eliminated.
`
`temperature. Remove excessive TFAC urder reduced
`
`Synthesis of His-Trp-Leu-Gln-D-Leu-Lys - Pro- Gly-Gln-Pro-lVlet
`
`pressure. Add 7.1N HCl/dioxane (0.19 ml, 1.32 mmol)ard then
`
`-Tyr:
`
`ether under cooling to make powder. Dry the powder over
`
`Dissolve Boc-l-lis-TrpLeu-Gln-DLeuLys (ZoCl)- Pro- Gly-Gln-
`
`NaOH for2 hours urder reduced pressure.
`
`Pro-lVlet—Tyr(2, 6-Clz-le)—Ole 000 mg, 0.3 mmol) in
`
`Dissolve this powder in DMF (10 ml). Add Boo-His (Tcs) (585
`
`hydrogen fluoride anhydride (15 ml) in the presence ofanisole
`
`mg, 1.43 mmol)ard then HOBT(193 mg, 1,43 mmol)tothe
`
`(1.4 ml), DMS (0.7 ml) ard ethanedithiol (0.7 ml). Alter stirring for
`
`solution. Subsequently, add WSCI (0.26 ml, 1.43 mmol) urder
`
`60 minutes at 00C, remove excessive hydrogen fluoride
`
`cooling, ard stirforone hour urder cooling and for 15 hours at
`
`anhydride urder reduced pressure. Add etherto residue to
`
`room temperature. Pour the reaction liquid into ice cold water
`
`deposit solids. Alter washing well with ether, dissolve the solids in
`
`(500 ml)to make itpowder. Drythe powder obtained by
`
`50% acetic acid, ard pass through Dowex 1 x 2 (acetic acid
`
`reprecipitating with methanol ard ethyl acetate / ether over
`
`type). Concentrate the solution thus obtained urder reduced
`
`phosphorus pentoxided for24 hours urder reduced pressure.
`
`pressure to produce a crude product.
`
`Yield: 1 .3 g *
`
`* Underthe aforementioned reaction conditions, approximately
`80% ofthe T06 radical on the imidazole of the histidine residue
`
`The crude productwas purified by gel column chromatography
`
`in TFAC (10 ml) under cooling, ard stirfor 10 minutes urder
`
`using Sephadex LH-20 and partition chromatography using
`
`cooling ard for40 minutes at room temperature. Remove
`
`Sephadex G-25 as a carrier (solvent: n-butanol / acetic acid /
`
`excessive TFAC urder reduced pressure. Add etherto residue
`
`water (4 : 1 : 5)). The product thus obtained showed a single spot
`
`to n‘ake it powder, and dry itover NaOH for2 hours urder
`
`on cellulose thin layer chromatography ard paper
`
`reduced pressure.
`
`electrophoresis.
`
`Yield: 250 mg (52.3%).
`
`Elemental analysis: as C71H104N18 016 - 2CH3COOH - 5HZO
`Calculated values: C, 52.74%; H, 720%; N, 14.76%.
`
`Dissolve the powder in DMF (10 ml), ard neutralize the
`
`solutionwith TEA urderccoling. Add Boc-Trp-ONSu (289 mg,
`
`0.72 mmol), ard stirforone hour urderccoling ard for 90 hours
`
`at room temperature. Add HOBT (81 mg, 0.6 mmol), ard stirfor
`
`Analytical values: C, 52.79%; H, 7.28%; N, 14.63%.
`
`15 hours. Pourthe reaction liquid into ice cold waterto n‘ake it
`
`Amino add analysis:
`
`powder, and filter it Reprecipitate the powdervvith methanol ard
`
`(Hydrolysiswith 6N HCl/ thbglycolic acid for48 hours at 1 10C)
`
`ethyl acetate/ ether, ard drythe powder thus obtained over
`
`m 1.0
`
`1.0
`
`1.0
`X2
`
`1.0
`
`1.1
`1.0
`X2 X2
`
`phosphorus pentoxide for24 hours under reduced pressure.
`
`Yield: 1.05 g (43%)*
`
`Elemental analysis: as C109H137021 NZOSCE - 4H20
`Calculated values: C, 5.5%; H, 6.43%; N, 12.32%.
`
`Synthesis of peptide B:
`
`Synthesis of Boc-Trp-l-lis-Trp-Leu-Gln-D-Len-Lys (ZoCl)—Pro-
`
`Gly-Gln-Pro-lVlet—Tyr (2, 6-Cl2-le)—Ole:
`
`Add DMS (1 ml) and ethanedithbl (1 ml)to Boc-His-Trp-Leu—
`
`Gln-D-Leu-Lys (ZoCl)- Pro- Gly-Gln-Pro-lVlet—Tyr (2, 6-Clz-le)—
`
`Ole obtained above (used as a mixture). Dissolve the solution
`
`-516-
`
`

`

`Japanese Unexamined Patent Publimtion No. 853-1 19872 (5)
`
`Analytiml values: C, 57.62%; H, 6.39%; N, 12.27%.
`
`on cellubse thin layer chrorr‘atography and paper
`
`*Yield from Boc-Trp-Leu-GIn-D-Len-Lys (ZoC|)—Pro-Gly-G|n-
`
`electrophoresis.
`
`Pro-lVIet-Tyr (2, 6-Clz-Bz|)—OBz|:
`
`Yield: 200 mg (40.4%).
`
`Synthesis ofTrp-l-lis-TrpLeu-GIn-DLenLys -Pro-Gly-G|n-Pro-
`
`Elemental analysis: as CgZH114N20 017 S - 2CH3COOH -
`
`Met-Tyr:
`
`5H20
`
`Dissolve Boc-Trp-l-lis—Trp-Leu-GIn-D-Len-Lys (ZoC|)—Pro-Gly-
`
`Calculated values: C, 55.80%; H, 7.02%; N, 14.79%.
`
`GIn-Pro-lVIet—Tyr (2, 6-Clz-Bz|)—OBz| 000 mg, 0.3 mmol) in
`
`Analytical values: C, 55.87%; H, 7.07%; N, 14.65%.
`
`hydrogen fluoride anhydride (15 m|)inthe presence ofanisole
`
`Amino acid analysis:
`
`(Hydrolysiswith 6N HCI/ thbglyoolic acid for48 hours at 1 10C)
`(1 .4 ml), DMS (0.7 ml) and ethanedithbl (0.7 ml). Alterstirring for
`60minutesat00C,remove exoess'we hydrogenfiuoride m
`
`anhydride underreduced pressure.Add ethertoresidueto
`>12
`)1;
`2523 w
`X
`deposit solids. Afterwashing wellwith ether, dissolvethesolids in
`Antimicrobial adion:
`50% aceticacid, and pass through Dowex 1 x2 (aceticacid
`T the
`ed'
`2 Iof
`i 8:208 nt
`so;corscenltirgfiérEOf Pies:
`type). Concentratethesolutionthusobtained underreduced
`pe
`000M ng
`pressure to produce a crude product
`prese
`ThedudeWm purified by96'“tum“WWW gtmmmmbzmg
`
`using Sephadex LH-20 and partition chromatography using
`
`'
`
`'
`
`tod xtrosemed'
`
`ddedvvith
`
`Sephadex 625 as a carrier (solvent: n-butanol / acetic acid /
`
`water (4 : 1 : 5)). The product thus obtained showed a single spot
`
`O: Peptide that showed antimicrobial adion throughout the
`period.
`A: Peptide that retained antimicrobialadionforoneweekorso.
`x: Peptide thath notshowantimicrobial adion.
`Anibaoterialteetforpepidek
`
`Strains
`
`Concentration of peptide
`
`Flavobaderium
`BaCiIUS megaterium
`BaCIIIUS ogagulans'
`
`O
`O
`O
`
`O
`
`
`
`
`><><><><><><><><><><><><>>>>>>>
`
`0
`
`O
`O
`O
`
`O 0
`
`
`
`GO
`
`0
`
`><>><><><><><><><D
`
`O
`
`GO
`——“
`—m-——
`OO
`Fusarium moniliforme
`Cucumber scab pathogen
`
`Potato anthracnose
`Mpergillus oryzae NRRL692
`
`Elisinoe ampelina
`
`—n——
`—n——
`—nnu
`—nnu
`—nnu
`O
`—nnu
`O
`%eudomonas Taesrroliteica*
`Q
`Q
`A
`Rice sclerotium hydrophilum
`
`Trid'tophyton mentagrophybe
`
`Sweet potato Fusarium
`
`GOOD
`
`(* Translators note: only pl‘onetimlly equivalent. I could find nowhere including Web sites.)
`- 517 -
`
`OO
`
`

`

`Antimicrobial test for peptide B:
`
`Japanese Unexamined Patent Publimtion No. 853-1 19872 (6)
`
`O
`
`O
`
`Bacillus coagulans
`Brevbaderium amoniagenes
`Cellubmonas oellulase
`Pseudomonas fluorescens
`
`Rice leafsheath scab pati'iogen
`
`0
`
`COD
` OOOOO
`OOO
`
`><>< 0! ><><><><><><><><><><><><>>>>
`
`
`H><><><><><>OOOO
`
`Rice (illegible) pathogen
`
`Cucumber scab pathogen
`
`Potato anthracnose
`
`Elisinoe ampelina
`
`Sweet potato Fusarium
`
`Fusarium oxysporum
`
`Gibberella fujikuroi
`
`Wheat blight pathogen
`
`Cochriobolus miyadeanus
`
`Magnaporthe grisea
`
`Rice sclerotium hydrophilum
`
`O
`
`><>
`
`—“
`—n——
`—nu—
`—n——
`—nnu
`—m---
`
`—nnu
`—nnu
`—nnu
`—nnu
`—nnu
`
`Bacillus megaterium
`Q
`Q
`A
`
`(* Translators note: only pl‘onetimlly equivalent. I could find rowhere including Web sites.)
`
`Applicant: Suntory Umited
`
`Agent Toshio Takigawa
`
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