406 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /., No.3,.*0.++ (,**1) 18 KFP. * * * * Sporulation of Bacillus subtilis (natto) KFP. Isolated

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1 406 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /., No.3,.*0.++ (,**1) 18 KFP. * ** *** * ** *** Sporulation of Bacillus subtilis (natto) KFP. Isolated from Takahashi natto Starter Derived from Food Extracts Saori Mitsuboshi, Rie Obitsu, Mayu Otsuki, Mina Katsumata, Rieko Kita, Sugio Watanabe*, Tadayoshi Tanaka**, Kanako Muramatsu*** and Kan Kiuchi Faculty of Home Economics, Kyoritsu Women s University,,,+ Hitotsubashi, Chiyodaku, Tokyo +*+2.-1 * Suzuyo Kogyo Co.,,33 Oyamahigashicho, Itabashiku, Tokyo +1-**+. ** Department of the Science of Living, Kyoritsu Women s Junior College,,,+ Hitotsubashi, Chiyodaku, Tokyo +*+2.-1 *** Department of Human Life and Environmental Science, Niigata Women s College,.1+ Ebigase, Higashiku, Niigatashi, Niigata 3/*202* We investigated the sporulation of vegetative cells of Bacillus subtilis (natto) KFP., isolated from commercial Takahashi natto starter. When the temperature of culture plates was increased from -1 to.*, the spore number (/ml) after sporulation treatment was observed to increase, and the number of spores was maximized by using cells from the second culture day. In a previous study, the addition of *.*+ mmol/ lmn, to the culture medium increased the spore ratio (ratio of spore number to whole cell number) from 1, (data obtained in a previous study) to more than 3*. In the present study, as inorganic Mn, was not permitted as a food additive, we prepared extracts from four foods with high Mn, content : dry sea lettuce, raw and dry green laver, and poppy seeds. By adding these extracts to the medium we could achieve a spore ratio of more than 3*. There were no large di#erences in organoleptic test results for natto produced with starters prepared from four kinds of food extracts. (Received Nov. +,,**0 ; Accepted Jun. +/,,**1) Keywords : Natto, Bacillus subtilis (natto) Takahashi, Sporulation, Manganese, Food extracts : +, *+2.-1,,+ * +1-**+.,33 ** +*+2.-1,,+ *** 3/*202*.1+ Corresponding author takahashi@s+.kyoritsu-wu.ac.jp *.+ mmollmn,. - 3* KFP., /

2 19 : *+, Bacillus subtilis natto KFP. Mn, *.+ mmoll *.*+ mmoll Mn, Mn + Bacillus subtilis natto KFP. KFP. +-, NBP NaCl *./, Beef extract +, Phytone Peptone + +., +,+ +/ Mn, NBP MnSO. *+.* mmoll NBP - NBP NBP 2 ml -** rpm *.*2 ml NBP 2 ml -** rpm -1 -*. +/ +*.* g,** ml +/* g +** ml +*.* g +** ml +,+ 0* -***rpm, +* /. /* l NBP -1.* +. +*.* ml +* *** rpm, +* +, *** rpm, +*, +*.* ml.,. +*.* ml,/g.*.* +*.* ml 0 1 SY-No.,* +* - + g +0 +1,* ;, ; - ;. ; / ; 0 ; 1. X x X SPSS Scheffé +2 2 Mn, Mn, RQ- Flex E. Merck 3 RQ-flex RQ-flex.,/2/* m, +

3 408 /. 3,** Fig. + Spore numbers obtained from cells cultured at -1C or.*c in NBP media and its spore ratio, -1C ;,.*C. KFP. was cultured at -1 or.*c on NBP plate culture with *.+ mmol/l Mn,, and cells obtained were treated for sporulation. The data show the number of spores per ml and its spore ratio. Fig., Relationship of concentration of added manganese with spore numbers obtained from cells cultured at.*c in NBP medium and its spore ratio, No addition ;,*.*+ mmol/l ;, *.+ mmol/l ;, +.* mmol/l. KFP. was cultured in NBP medium with *, *.*+, *.+ or +.* mmol/l Mn,. The data show the number of the spores obtained after sporulation and its spore ratio. moll HCl + moll NaOH + KFP. -1 Mn, *.+ mmoll NBP -1.*. Fig * 1 ml,.,./+* 2 ml-.,+* 2 ml 2*.* * 2 ml, 2.,+* 2 ml, -..,+* 2 ml,. -.3+* 2 ml -1.* 3* 33+**.*, KFP..* +, -1..*, Mn, Mn, NBP Mn, * *.*+ *.+ +.* mmoll Fig.,.*. Mn, *.*+ mmoll, Mn, *.*+ mmoll +.++* 3 ml 32+** Mn, *.+ mmoll + KFP. Mn, *.*+ mmoll NBP.*,,.1 *.+ mmoll / KFP + KFP, *.+ mmoll

4 21 Table + Concentration of Mn, contained in their extracts : 409 Sample Dried sea lettuce Raw green laver Dried green laver Poppy seed mg/l of extract *420 +4*. *403 *40, Extracted Mn (mg) /+** g of sample +4/- * ,+ *4/11 Samples were bought at The Wholesale Market at Tsukiji, Tokyo4 KFP. *.*+ mmoll, 1/.2 KFP. *.+ mmoll + Mn, Mn, - Mn, Mn, Mn, Mn, +/ +** g +1.** mg, +-.** mg, 0.22 mg Mn Mn, Table + +**g Mn, Mn, *.*+*.*, moll Mn,**/ Mn Mn Table + Mn. NBP KFP. Fig. - Fig. - A *.*+ mmoll Mn, +.++* 3 ml,.-+* 2 /ml,,.+ +* 2 /ml, *.3+* 2 /ml, *.2+* 2 /ml B 33+** Mn, Fig. - Spore number obtained from cells cultured in media added food extracts and its spore ratio MnSO. *.*+ mmoll Fig., Mn, Mn, / Table, 1 p*.*/. X p*.*/ X p *.*/ p*.*/ X. p*.*/ X X p*.*/ *.*+ mmoll Mn, p*.*/ Bacillus subtilis natto KFP. -

5 410 /. 3,** Table, Organoleptic test of natto made with starter of spores prepared with food extracts Item Dried sea lettuce Raw green laver Dried green laver Poppy seed *4*+ mmol/l Mn, Takahashi starter Commercial natto Appearance Lysis of bacilli Color of soybean Crack of soybean Stickiness Smell Di#erent odor from natto Toughness Umami Taste of soybean Bitter taste Total evaluation Acceptability +42*4/ 04*+4, a,4**4* ab.4-*4/ +4-*4/ bc +42+4* c.4-+4* a.42*4/ ab,4**42 c +42*4/ b.4/+4- +4/*40 c ab +42*4/ /4/+43 ab,4-*4/ b.4-*4/ +4/*40 bc,4/*40 c -4/+4- ab /4**4* ab,4**42 c +42*40 b -42+4/ +42+4* c,42*4/ b,4/*40 /4/+43 ab,4-*4/ b.4/*40 +42*40 bc,4-*4/ c.4/*40 a /4**4* ab,4**42 c +42*41 b * c,42+4* ab,4/*40 /4*+4. ab,4-*4/ b.42*4/ +4**4* ac bc -4**42 b.4/*40 b,4/*40 c,4/*40 b -42+4* +4/*40 c,42*4/ b,4-*4/ /4*+4. ab,4-*4/ b.4/*40 +4/*40 bc -4**4* bc -4/*40 ab.42*4/ ab bc,4/*40 b -42+4*,4**42 bc,4/*40 b 04-+4* a 04**4* ab -4**42 ab.4/* bd 042*4/ a,42+41 ab /42*4/ a /4/+41 a.4/*40 a -42*4/ -4/+4- ab,4/+4* ab.4**4* b.4**4* b.4**4* a.4**4*.4**4* ad.4**4* b.4**4* ab.4**4* b.4**4* ab.4**4* a.4**4*.4**4* a.4**4* a Data are presented as means4d4 values (n3)4 Assessment was scored on a scale of + to 1 (+far inferior,,inferior, -slightly inferior,.equal, /slightly better, 0 better, 1far better) compared to commercial natto as the control4 a, b Mean values with di#erent superscript letters in the same row were significantly di#erent at p*4*/ (Sche#é post hoc test)4 - KFP.. *.*+ mmoll Mn, Bacillus subtilis KFP. Mn, -1.*, Mn, *.+ mmol l *.*+ mmoll 33./ Mn, NBP KFP. 3* *.*+ mmoll Mn,. Mn, +,, pp. 30 +** +3.3, Kiuchi, K. and Watanabe, S., Industrialization of Japanese natto. In Industrialization of Indigenous Fermented Foods, Second edition, Revised and Expanded, ed. by Steinkraus, K.H., (Marcel Dekker), pp. +3-,.0. (,**.). - Hosoi, T. and Kiuchi, K., Natto, a food made by fermenting cooked soybeans with Bacillus subtilis (natto), In Handbook of fermented functional foods, ed. by Farnworth, E.R., (CRC Press), p.,,2,/* (,**-).. /- +0/+1+,**0 /.+ +,-+, ,

6 23 : 411 1, 3 +3-/,**- 2 Kim, H.-R., Muramatsu, K., Kanai, Y., Tanaka, T., Takeyasu, M. and Kiuchi, K., Development of a new soyprotein fermented food employing chungkuk-jang starter, Food Sci. Technol. Int. Tokyo, -, -+*-+0 (+331). 3 / ,**0 +* B +, -/ +/. +/ , /1/ /2, +33/ +,.2,21,32,**+ +- /* +2, Joko, S., Taya, N., Funane, K. and Kiuchi, K., Production of natto starter. Nippon Shokuhin Kogyo Gakkaishi, -/,,12,2- (+322). +/ p. 0,0- +-*+-/,*** +0.2,11,20,**+ +1 p * +2 SPSS Inc., SPSS Base +,.* J User s guide pp. -*3-+/,** /

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 Starter of Bacillus subtilis KFP 2.- Isolated from Douchi