454 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /,, No.+*,./..0+ (,**/) 20 Bacillus subtilis KFP 2.- * * * * Soft Natto Manufactured with Star

全文

(1)454. Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /,, No. +*, ./.º.0+ (,**/) »/3¼. ¹ 20 . ῌ Bacillus subtilis KFP 2.- .

(2)

(3) ῌ

(4) *

(5) !"#** $ %*** & ' () *+,* * - () ./*012*2 ** (./*012*2 *** 345678. Soft Natto Manufactured with Starter of Bacillus subtilis KFP 2.- Isolated from Douchi Saori Takahashi, Rie Katsumata, Kumi Yoshizawa, Momoko Yamaki, Miyuki Osako, Mariko Murakami, Sayaka Mouri, Mizuki Sato, Hiroko Meguro, Hiroko Kubokura*, Kanae Hirose, Tadayoshi Tanaka**, Sugio Watanabe*** and Kan Kiuchi Faculty of Home Economics, Kyoritsu Womens’ University, ,ῌ,ῌ+ Hitotsubashi, Chiyoda-ku, Tokyo +*+ῌ2.-1 * Presen address : Department of the Science of Living, Kyoritsu Womens’ College, ,ῌ,ῌ+ Hitotsubashi, Chiyoda-ku, Tokyo +*+ῌ2.-1 ** Department of the Science of Living, Kyoritsu Womens’ College, ,ῌ,ῌ+ Hitotsubashi, Chiyoda-ku, Tokyo +*+ῌ2.-1 *** Suzuyo Kogyo Co., ,3ῌ3 Ooyamahigashicyo, Itabashi-ku, Tokyo +1-ῌ**+. ῍῍῍῍῍῍῍῍῍῍῍῍῍῍῍῍ We collected non-salted Douchi from Yunnan Province, China, and isolated the dominant bacteria for the production of Natto. We identified the most predominant bacteria as Bacillus subtilis, called it KFP 2.-. The toughness of Natto made by us with this strain was about .*ῌ of commercial Natto. Formol nitrogen content and relative viscosity of Natto made by us with B. subtilis KFP 2.- was about -*ῌ and /*ῌ, respectively, of those of the commercial Natto. Organoleptic assessment revealed that the stickiness of Natto made by us was weaker than commercial Natto, but that the former was softer. The appearance, clack of soybean and toughness were considered good. The softer Natto will be useful for individuals who experience di$culty swallowing or in mastication. (Received Jan. ,2, ,**/ ; Accepted Jul. 2, ,**/). 9:;<=>?@ABCDEFGHIJKLMNO PQHR: =>?@<NDSTGH OS(D. AFYlGH^ 'DA a<"o:PQ` 9:;<. U<VWXY=>DTZ[\HZ<]:=. {|"oA!;oῌ<(D_oῌD\RT<M. WXQHR^ =>_`MGabc<dGeM. R+^ !;oῌA{|"o<zDoE . :kilmQH. <OP WYD 0)EOQlmR. R^ 9:;<n!"oApqr: sP #tu$:%v. Zf:gGh<?<YiDTj. : H^ }SPQ{E|aT<AGa T. \RDwmQxyz`T&lH{|"o:}~mRz. Q *T< ¡+¢£<,G-GeR?Dz`. DGXQY: WU<VH><Yi<"o. ,¤M 9:;<"oSA, :¥GXQHR^ ¦¤A!;§'¨MoῌE©ªPQ «5E(.¬P. ±+*+ῌ2.-1 ²³ ´hµ ,ῌ,ῌ+ * ±+*+ῌ2.-1 ²³ ´hµ ,ῌ,ῌ+ ** ±+*+ῌ2.-1 ²³ ´hµ ,ῌ,ῌ+ *** ±+1-ῌ**+. 0´¶· ,3ῌ3 12¸ ¹Corresponding author. "oE}~PY® <!D 9:;<n!"oz`T &lao<w¯m\H{|"oE}~PR«5EF HPY<M/°\R^.

(6) ῌ¼ῌ :

(7) +,. 21 . 455. + . ῌ KFP 2.-.

(8)

(9) Bergey’s Manual of Systematic Bacteriology Vol. ,, Bergey’s Manual , . Marble ../ /.1 mm. mm. ! ,**+ ". -

(10) . # NBP NaCl *./῍, Beef extract. Fig. +. Douchi collected at Wayao in China. +῍, Phytone Peptone +῍ $ NBP %&. ".. Wxb ῌ 3

(11) T kT n

(12) $q. . NBP %& '(). ! *" #$. %&

(13) ' +(#*" )/. xb $T Gopqrs4 / 2' KFP 2.- x. / . ,*+,-. /0-. + SY-No. ,* . , (.

(14) 1234 /02 +,156. Table + KFP 2.- . ") 2343 567+89:;<=>1. P

(15) t *.2 mm u vw x. 57

(16) . y + zx Bacillus {{. /. 0 . !. ?@ A9BC985 9:;")D<E. x @ b

(17) 1῍ | Nutrient broth # w }~

(18) z. FGH40I3 := . <LC7u

(19) !# V-P. " A J+ a KL6M> GH"N?. broth '\ pH 0 1 ^xb Dῌ |9C. " !O@ ?@) PA

(20) A'QRB S TURV UCWXDE g F. 7 LῌL ¡C7 Dῌ¢£zC7 Dῌ{6;uC9z 1 Dῌ|9C787 =6 L¤6 ¥6>6%¦

(21) y§61¨©

(22) . YG )ZH[CI\ /] ,. X^J.

(23) ¤z£6¦ª

(24) t«;9L;6¬¡. _ `* + X^K.a

(25) O@a. ª

(26) ®7uª

(27) 1¯°

(28)

(29) . L`* + X^K.\). =6MC9ª

(30) pH 0 $ 2 Nutrient broth. D<EF Mb NOP Q c3ῌ. #

(31) Bergey’s Manual ±²' . RV3 def gb @ R

(32) Sh i. 2 +*] ³´ /*] # Bergey’s Manual. ?@ jY T UYῌkY VlWm Xn +- h. d ++ῌ23῍

(33)

(34)

(35) . YR

(36) O@ Zo . 1 ZoWm + [\. +*] #SQ /*] #µ<x. p

(37) ; , p

(38) ; - qqp

(39) ; . ]^ ; / qq_

(40) ; 0. KFP 2.-

(41) Bergey’s Manual . . _

(42) ; 1 [\_

(43) `rab SPSS J. ±²@3 Bacillus subtilis { x. É vwLC stu+* " Scheffe. 2 KFP 2.- B. subtilis KFP 2.- ±. ,* cd

(44) ++ ex. . 1 "#$ !. - ) . f34g87yz{u|Lt}Ch. +,H*" +(*"H[ H[. . *" ¶

(45) ·/ B. sub-. +++,. % &. tilis KFP 2.- +(#*" .-] x 2. +,*">z|L¸ ¹ Fig. , B.. + '. subtilis KFP 2.- +(#*" .-] 0 º *ῌ. ZG~ῌ Fig. + . 2 X^Y .-] +*ῌ+- X^Y .2] +/ῌ,* X^Y. ῍a ijkqlmn . .-] 2\ ,* X^Y*"» t !¶ ,..

(46) ±῍²p³´µ´¶· ¸ /, ¹ ¸ +* º ,**/ » +* ¼. 456 Table +. J 22 . ῎ /

(47) ῌ. The characteristics of KFP 2.-.

(48)

(49) . !"#$%. KFP 2.-. Bacillus subtilis*. Gram’s stain Spores heat-resistance test Cell diameter Spores round Sporangium swollen Catalase test Anaerobic growth Voges-Proskauer test. *42 ® ® ® . *42 ® ® ® . pH in V-P broth¯0 °1. ® ®. d ®. P)<=>?@ῌ

(50) MQ<=. Acid from D-Glucose L-Arabinose D-Xylose D-Mannitol. . . Fig. - <=>?@ .- R .1

(51) HS&. Gas from Glucose. ®. ®. Hydrolysis of Casein Gelatin Starch. . . Utilization of Citrate Propionate. ®. ®. Degradation of tyrosine Deamination of phenylalanine Egg-yolk lecithinase Nitrate reduced to nitrite. ® ® ® . ® ® ® . Formation of Indole Dihydroxyacetone. ® ®. ® ND. Growth at pH 042, nutrient broth /41, nutrient broth. . . Growth in NaCl /ῌ 1ῌ +*ῌ. . ND. Growth at / +* -* -1 .* /* // 0/. ® ® ®. ® d d ® ®. Growth with lysozyme present. ®. d. Characteristics. * The characteristics of Bacillus subtilis are those described in Bergey’s Manual of Systematic Bacteriology Vol. ,. Symbols are as follows : (®), 3*ῌ or more strains are negative ; (), 3*ῌ or ,more strains are positive ; (d), ++῏23ῌ of strains are positive ; (ND), No data available.. &'" !"()*+,+ . -..

(52) /012)34#$

(53) ῔345678 9

(54) :;#$"<=>?@ABCῒDE F G&" .- <=>?@ <=>?@ .- HI

(55) #$E34 KFP 2.- J.- 34 HKELHE MN. :;"<=>?@()O . -2" #$E<=>?@ <=>?@ .1 HI

(56) #$E 34 KFP 2.- J.1 34 HKELHE * >?@ῐ2 'TUVH34῏WVXYZ[\DE &'T ῏WVXYZ]^_ELH34`]ab

(57) c& &'

(58) 2LHDE <=>?@ .- &'de 0 ῎6XYbf]g

(59) .( ῏WVXYZ +* ῎ /ῌ hi( ,* ῎Rjk

(60) <=. >?@ .1 0 ῎6XYbf]g

(61) .( 3 ῎6 +3 ῎RXYZ] /ῌ hili

(62) ] *+hm῏WVXYZ] /ῌ h::n

(63) . . o"<=>?@#$

(64) #p34qrῌ

(65) stΐu4vῐwx34H89

(66) JFig. . wx3 4y7 ,/* g z{ 0** g hi J|}~ -3-1*.g *+

(67) <=>?@ .- #$

(68) . y7 /* g z{ -** g hi J|}~ +.3.-.. g <=>?@ .1 +** g z{ ./* g hi J|}~ ,2+1*.- g .- ()y234.

(69) 6

(70) 34)wx348H56 . <=>?@ .- #$EH56234] S /

(71) ῌ Y Table , DE Y῕wx34. ] *.3.ῌ

(72) KFP 2.- J.- 34*+ *.-2ῌ KFP 2.- J.1 34/0 *.,2ῌ H 2+. ) .]R(22LHD

(73) B. subtilis KFP 2.- #$

(74) 34Y῕. &'"wx34*+()l6 ῍ . 56234 Table - DE wx34 ] ,.*0

(75) KFP 2.- J.- 34 +.+*, KFP 2.- J.1 34 +.,+ KFP 2.- J.- 34] - pT)l6 ῎ &. T ¡?¢£¤¥=¦§>?¨©H GC-MS 34&ª

(76) 34«ῑ¬.

(77) ῌ¨ῌ : UO9:;Va. 23 . Fig. ,. 457. Temperature and humidity programs for manufacturing natto with Bacillus subtilis KFP 2., Temperature program .- (initial temperature is .-8) , Temperature program .1 (initial temperature is .18) , Humidity program. +* ++

(78) ,-῍ ,῍! -῍"#$%&῍,῍ '( ,/῍ ! ,-/῍')! ,῍! *$+, (-., -῍!., (-/0, ,῍ !.,123

(79) 45617 KFP 2.- .-8 9:;<=>?@ Fig. /

(80) A;BC KFP 2.- .-8 9:D7 +* E ;F;G ,-῍ -῍! ., (-/0, ,῍!.,; - H;IJK LMNOP

(81) KFP 2.- .18 9:;<=D7 KFP 2.- .-8 9:;ANQRS7TUNOP

(82) ῌ VW;XYKZ Table . D[\*$] ^_` B. subtilis KFP 2.-. 1Va 9:;XYbc;BCd

(83) KFP 2.- .-8 9:7 Vaefg;hijhkî R: ;l7mNnUo pq7rsMN39:tu vwxOP

(84) yz7{{|yzJ :;}N{H ~N7OP

(85) KFP 2.- .-8 9:7 DUO OR fD:JH~N 2 O ;GDP. Fig. 0

(86) AN7DQ: zH~ Gz. Q:9:;z. G12P

(87) R7OP

(88) KFP2..-8 9:7 :;}NῌH~N M;1; , H. ;9:DD p*.*/ bcJN

(89) MU D DH7rs9: :;¡DH 7 KFP2.- .18 9:D p*.*/ bcJ ¢OP

(90) KFP2.- .18 9:7 ;£zJ; , H z D p*.*/ bcJN

(91) ¤¥;1 7 KFP 2.- .-8 9: rs9:D KFP 2.-. Fig. -. Oxygen concentration during manufacturing Natto with B. subtilis KFP 2.-. (A) Natto manufactured by using the temperature Program .- shown in Fig. , (B) Natto manufactured by using the temperature Program .1 shown in Fig. , ¦, O, concentration in the chamber §, O, concentration in the natto package.

(92) ¯oM°±²³ ´ /, µ ´ +* ¶ ,**/ · +* ¸. 458. Fig. .. 24 . Toughness of three kinds of Natto. The three markings of poles show the using temperature programs for natto manufactured and commercial natto as follows : , Temperature program .- ; , Temperature program .1 ; ®, Commercial natto.. Table ,. Formol nitrogen concentrations in Natto Natto. Table -. Formol nitrogen (ῌ). Natto + Natto , Commercial natto. *4-2 *4,2 *43.. Relative viscosity of Natto. Natto. Relative viscosity. Natto + Natto , Commercial natto. +4+* +4,+ ,4*0. Natto + and , were made with B. subtilis KFP 2.- using the temperature programs .- and .1, respectively, shown in Fig. ,.. Natto + and , were made with B4 subtilis KFP 2.- using the temperature programs .- and .1, respectively, shown in Fig. ,.. .1

(93) . pgFuz =>y` oM- =. !"#$%& '(. \= {| pg$ ^> bc+$. ) *+, )-. }~

(94) *ZZ[\= y-. ῌ. ῍. ῌ\

(95) D =_ KFP 2.- :!= ?DIYGH\

(96) B. subtilis $+y^. ./01213$45 6789191. - /$@A > _ ./. :;<=>?

(97) :@ >:'(=> 0121. _ !GH:y . B. subtilis. A > BCD8919101. natto $ m

(98) , /?_:@. 21 - 589191:@ >'(A > - ?E. >B:<Ay$. F0121GHIJ=>KL M. 1ῌ : m>¡¢=y,. NOP:+:QRSTUVWXY. U. o. :£=+- B.. subtilis KFP 2.- y!$ _ +.¤ Bacil-. $'(+Z[\= - ]^> ./_Q. lus subtilis natto. R`La

(99) \ bcdef gE. ¥+,. !GH:+y. )- ῌ\

(100) D =¦_y! B.. :'(+hijk$ lm- n\

(101). subtilis natto $ GH:y, ῌ /. oA > E$%)pg$oqr >. !§¨©B<oM, :ª«¬. oML stBFuHvwx y. y-.

(102) ῌ¼ῌ· : -32!. 9 25 ;. Fig. /. 459. Chromatogram of volatile components in Natto made with B. subtilis KFP 2.-. The components with peak numbers in the chromatogram are as follows : No. +/, ethanol ; No. 0*, pyrazine ; No. 1-, ,-methylpyrazine ; No. 1/, --hydroxy-,-butanone (acetoin) ; No. 10, ,,/-dimethylpyrazine ; No. 22, ,,-,/trimethylpyrazine ; No. +*+, ,-methylpropanoic acid (isobutyric acid) ; A (No. ,+), ,,--butanedione (diacetyl) ; B (No. +*1), --methylbutanoic acid (isovaleric acid) ; C (No. +*2), ,-methylbutanoic acid.. Table .. Organoleptic assessment of Natto made with B. subtilis KEP 2.- using temperature programs .- and .1 Commercial Manufactured natto natto Program .- Program .1. JKL ECMM HI/"C!# 2 HIN!OP QRSK"H T" U!!V"# I WXY! Z[.\]T ^K"/_H+.; àbcdef , ghi". 04*¹*4*a /42¹+4+ab -4*¹*4*ab .42¹+4+b +4*¹*4*b ,42¹*4. -4*¹,41 .40¹+4/b -4*¹+4, +42¹*4.b -42¹,4* .4,¹+4+ .42¹+40. jk WX/IDlmno+2. Data are presented as mean¹S.D. values (nº++). Assessment was scored on a scale of + to 1 (+ºfar inferior, ,ºinferior, -ºslightly inferior, .ºequal, /ºslightly better, 0ºbetter, 1ºfar better) compared to commercial natto. a,b Mean values with di#erent superscript letters in the same row were significantly di#erent at p»*4*/ (Sche#e ´ post hoc test).. & 3+,; A 0 g'Y! . . Appearance Lysis of bacilli Color of soybean Clack of soybean Stickiness Smell Di#erent odor from natto Toughness Umami Taste of soybean Bitter taste Overall estimation Fancy. .4*¹*4*b .4*¹*4*b .4*¹*4*a .4*¹*4*b .4*¹*4*a .4*¹*4* .4*¹*4* .4*¹*4*b .4*¹*4* .4*¹*4*a .4*¹*4* .4*¹*4* .4*¹*4*. -42¹*4.b 04*¹+4.a ,4.¹+4+b 040¹*43a +4*¹*4*b -4.¹+4/ .4*¹+43 14*¹*4*a .4,¹+40 /4*¹+4,a ,42¹,4. -40¹+43 -4*¹+40. $XpACDE B. subtilis KFP 2.- \]TN2

(103) qr s!!C"E78+# t!uv \]T"w!EBlmC"xV3" 2TaKBy23# !M \Y z{P+" CD|}\3! D~E|/"xV3# \ ]T$/" \]T5I!!"

(104) qK"OP WXM "%KT/"# !+"E( ) !"# .*cde? ef)+,; B"+( T ! B. subtilis KFP 2.- 23 , u+E 3 !" ,ῌ=, -ῌ=, u+2# A. B. subtilis KFP 2.- !

(105) . !k ¡3 \]¢ k £V!E/#. !"# ,* $% &' ()

(106) . ¤ ¥ -32!K"0AQ?. *+ PSP ,-./"01

(107) 23. Q? .¦

(108) §¨

(109) q. 456- 0/"78+"#. ¢!©K T2# ªZ«D/T¬D. KFP 2.- 9.-:; <=>?=@ 4 A. Z! -32!®BEC!. BCDE2# KFP 2.- 9.-:; . ¯° 032# Z1!"Q±²r. KFP 2.- 9.1:; CDEF!G# HI. ³´M µ±>¶µB·!JMD ¸2.

(110) ³Va´kµk¶· ¸ /, ¹ ¸ +* º ,**/ » +* ¼. 460. Fig. 0. 26 . Photographs of Natto made with B. subtilis KFP 2.- and commercial Natto (A), Before agitation of natto we made ; (B), Before agitation of marketed natto ; (C), After agitation of natto we made ; (D), After agitation of marketed natto..

(111) ῌ

(112) . y*+ zH-6{|}" . B. subtilis KFP 2.- . Q,&')

(113) *+H-@68-! KFP. !"#$% &' ( )

(114) . 2.- /0 37 KFP 2.- . Bacillus. *+,-!./0 KFP 2.- .1234. subtilis "~H-!G .

(115) . ."5 B. subtilis 6378 *+9 . ,. B. subtilis KFP 2.- *+" , . 6 KFP 2.- .6. KFP 2.- .eG-. :;<"=> ? !-@6"AB. .- c6H-*+ B *+. CD " )

(116) <5 EFG-. ;< ;< +**῍ 6H-6 %. *+H-@68- B. subtilis KFP 2.- -6. .*῍ ;<"=> . I) J -

(117) 6,@6"5KLMNOP. -. . % *.3.῍ G. GB Q RS"T6-U"V6WX. " !.*+ *.,2 *.-2῍. ,@6YTG-6Z[-. G Qxy8 AB\\^J . )

(118) \ ]^_`Va6 bc8 dQ0e[ fg\ h J)

(119) VaT6H- gVa6. % ,.*0 G " +.+* +.,+ G . K !B ῌ5 ;<.

(120) VaGi, MN< ")

(121) j. . klmnH-@6"A opVa6Xq. G "#$

(122) " p*.*/ xy8^. erJ-Gi,. )

(123) 6,WX. ῍. ῌ. + stuvῌ

(124) w!x. "#$

(125) "WX p*.*/ . ¡R¢H-"GB R£"¤¥08Q ¦§¨B©ª«¬®©"¯J°±>²QH.

(126) ῌ¨ῌ© : ª« IJ. 27 . ῍. ῌ. + .

(127) + .. ,+0ῌ,,* +330 , Claus, D. and Berkley, R.C.W., Genus Bacillus Cohn +21,, +1.AL. Bergey’s Manual of Systematic Bacteriology, Vol. ,, Eds. by Sneath, P.H.A., Mair, N., S., Sharpe, M., E. and holt, J.G. William & Wilkins Co. (Baltimore, U.S.A.). pp. ++*/ῌ++-3 +320 - Sulistyo, J., Taya, N., Funane, K. and Kiuchi, K., Production of natto starter. Nippon Shokuhin Kogyo Gakkaishi, -/, ,12ῌ,2- +322 . !" #$%" &'( )*+" ,-." & / 0 123456789 :; <= ., /1/ῌ /2, +33/ / !" >?@A BCDE -FGH &/ 0

(128) IJKLM <= .2 ,11ῌ,20 ,**+ 0 NOKPQR STUVW<XYZPQ[\ ]^ NOK _ à;b pp. +, +1 +33* 1 NOKPQRῌSTUVW<XYZPQ[ cde5 dfg NOK p. ,, +33*. 461. 2 NOKPQRῌSTUVW<XYZPQ[ hijk lmK NOK p. /2 +33* 3 NOKPQRῌSTUVW<XYZPQ[ nopq K NOK p. 0+ +33* +* SPSS Inc., SPSS Base +*.* J for Windows user’s guide, 1 3ῌr5ῌ13ῌ13_ st p. 1ῌ+. ++ Tanaka, T., Muramatsu, K., Kim, H-R., Watanabe, T., Takeyasu, M., Kanai, Y. and Kiuchi, K., Comparison of volatile components from Chungkuk-jang and Itohikinatto, Biosci. Biotechnol. Biochem.,0,, +..*ῌ+... +332 +, -FGH u/v" >?@A &/ 0 wxyhz{| }3~25

(129) ;9 <= /* ,12ῌ,2/ ,**- +- Kim, H., Muramatsu, K., Kanai, Y., Tanaka, T., Takeyasu, M. and Kiuchi, K., Manufacture of chungkukjang with elastase activity. Food Sci. Technol., -, ,/+ῌ,/0 +331 +. -F H ῌ ¡x ¢<=£ -0 302ῌ32* +323. +1 ¤ + ¥ ,2 ¢¦§ +1 ¤ 1 ¥ 2 ¢¦@. ῍῍῍῍῍῍῍῍῍῍῍῍῍῍῍῍῍῍῍῍῍῍῍῍.

(130)

454 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /,, No.+*,./..0+ (,**/) 20 Bacillus subtilis KFP 2.- * ** *** * ** *** Soft Natto Manufactured with Star

454 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /,, No.+*,./..0+ (,**/) 20 Bacillus subtilis KFP 2.- * * * * Soft Natto Manufactured with Star