Growth Inhibition of Yeast Isolated from Processed Rice Cake with Ethanol and Organic Acids

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1 J. Fd Hyg. Safety Vl. 22, N. 2, pp. 99~104 (2007),QWTPCN QH (QQF *[IKGPG CPF 5CHGV[ Available nline at Œ ò ƒœ æ ô Œ    ÂÞ * w t œw Grwth Inhibitin f Yeast Islated frm Prcessed Rice Cake with Ethanl and Organic Acids Jng-Shin Kim, Hyun Jun Lee, Yung-Tack Lee, Hak-Gil Chang, and Jng-Hyun Park* Department f Fd Science and Bitechnlgy, Kyungwn University (Received May 10, 2007/Accepted June 11, 2007) ABSTRACT T defend putrifactin f the prcessed rice cake frm gas-frming yeast during strage and distributin, it needed t reduce and remve them. The sanitizers f ethanl and rganic acids were applied n Pichia anmala, Candida trpicalis, and islated yeasts frm the putrified cut rice cake. Althugh grwth inhibitin effect by the sanitizer f 20% ethanl, 1% acetic acid, r 1% lactic acid respectively were very lw, the cmbined sanitizer f 20% ethanl and 1% acetic acid, r 1% lactic acid shwed very high sterilizing effect tward the yeasts. Six lg cfu/ml f the yeast was reduced with this cmbined sanitizers fr 30 minutes. In additin, the cmbined sanitizer heated frm 20 t 50 C had mre the increased sterility. Therefre, the sanitizer f the cmbined ethanl with the acetic acid r the lactic acid fr 30 minutes at 50 C might reduce r sterilize the putrifying yeast at the prcessed rice cake. The result might be als applied t the effective pre-treatment f many agricultural fd stuffs against yeast, especially unsterilized stuffs, withut any hazards frm the special sanitizers and nutritinal lss frm harsh sterilizatin. Key wrds: ethanl, acetic acid, lactic acid, sanitizer, yeast, prcessed rice cake õ y xk y w ³ ƒœ xk t m š. ù m, y ƒ t ƒ š ù y y w t p w ³ w w š t w ƒ. Bacillus, Clstridium s ³ ³ p û. w ³ ³ B. cereus t q j ù mx x š. ù x *Crrespndence t: Jng-Hyun Park, Department f Fd Science and Bitechnlgy, Kyungwn University, Sujeng-Gu, Sengnam-Si, Kyunggi-D , Krea Tel: , Fax: p5062@kyungwn.ac.kr Ê w w ƒ ƒœt wš. ƒœt š ƒœ t w ³ w» z š ³ w š. z ƒ x w s ƒœt t š ƒœ f ƒ. ù v» q w z w š š w w ³ w ƒ š. q y ³ w yw, hydrgen perxide, ptassium srbate, benzic acid 2,3), prpinic acid, citric acid, acetic acid, lactic acid» 4-7) NaCl 8) w x ƒ š š. w Š ³ w z š9)w š, w q w ù ³ ƒ š x ü ³ s š» w yw 99

2 100 J.-S. Kim et al. w 12) ù w š t w wš. w x wš t w w š w w»ƒ. q w w 13) ü w Bacillus w» w 14)ƒ k š y ƒ w w w ƒ. x ³ t y š k w³ z ú l ù t w w. k t s w 70% k t ³ š,,, t, k, t ƒ š. k ³ k ƒ w w w š ww w w ƒ w» ùkü». ³ 20 C, 5 ³ ƒ» k wz w 15). k 50% ƒ v w w» 1~2% w» 20% k 5 ü ³ z ƒ. š 15) ³ w k z šw ³ k y ww k 1 g w ³ y d w. k w w 50% ƒ ³ v w ù 8~10% ƒ w k 20% x w ³ ûš 10% ³ ƒ w. w k ƒw Bacillus cereus w ƒ š 16)» w. w w k, ƒ x, z y w,, w yk w». š 0.3% ptassium srbate 3% ƒ w»ƒ 12 š, ³ š w 15). ƒ w k» ww ƒ m jš ƒœt z w wš w. Ì Ý ßá ò Ý x t z ³ Pichia anmala ATCC 8168 Candida trpicalis ATCC 750 w. x z ³ t q s ƒœ ( w t,» ) l œ ³ w. 25 g ³ bag z 9 ³ (w/v) stmaching w z 25 C w 2 w ³ w. PDA (ptat dextrse agar, Difc, MI, USA) z w w w erythrmycin, chlramphenicl (Sigma, St.Luise, MO, USA) rse bengal (Sigma, St.Luise, MO, USA) ƒw w. g x y w z q w. k (Mallinckrdt Baker B.V, Hlland) lactic acid acetic acid (Duksan, Kynggid, Krea) GR 1 w z 0.22 µm filter unit membrane (Millipre, MILLER-GV, France) ³ w z w. GR w. ô ß œ z ³ Pichia anmala ATCC 8168 Candida trpicalis ATCC 750 q ƒœt l z w -70 C w w. ( ) w t t ƒ ƒ t t t l z w. ³ w w erythrmycin, chlramphenicl (100 ppm) w rse bengal (5%) ƒ YM (Difc, Detrit, MI, USA) w 25 C w 48 ¾ w. g w Ÿwx z y wš z w. ptat dextrse brth 10 ml 1% w 25 C 48 w š ³ w. ä Ð»Ï ô Œ» ƒƒ 1%(V/V), 3%, 5%, k 5%, 10%, 15%, 20% y w w z w ³ d w. š 10, 30, 60, 50 C¾ w z w z PDA w 48 w y w. t ³ ³ 1 ml ƒ 100 ml z

3 Grwth Inhibitin f Yeast Islated frm Prcessed Rice 101 w z PDA w 48 z y w. q ƒœt ƒ 10 v w z stmaching w PDA w wz ³ d w y w. ŠŒ Ý æ ½ä ƒœ ½ õ æð s ƒœ qw» ƒ ƒ s ü t š z w. ( ) w t t ƒ ƒ t t t l z w. ³ w w erythrmycin, chlramphenicl w rse bengal ƒ YM w 25 C w 48 ¾ w. g w x z y wš g xk 4 z w. 4 z Islate Y-1, Islate Y-2, Islate Y-3, Islate Y- 4 š w. ƒ g Islate Y-1 Islate Y-2 w w w. ô Ð Î ô k -» w z Pichia anmala Candida trpicalisƒ x w w z ƒ ùkû k - Islate-Y cfu/ ml w Islate-Y cfu/ml w w 10 5 cfu/ml w. ù k - p w z w ƒ z ƒ w (Table. ³ k ³ ³z ƒ t ³ y y š. ³ w» w š17) k peptidglycan ƒ w w ww š šw w w 15) k 8% 12%. w y k ³ w e ù k ƒ š. ³ ³ k 50% 10 w š 60% k 5 w ƒ š. ù ³ z k w z ƒ ùkû. w x y š k z ƒ w w š. k y p yww ƒœt» q³ z w j z ƒ ƒ. Ð»Ï ô Œ k ƒ 1%» w t z ³ z w z» w» 1% k 5, 10, 15, 20%ƒ w w 100 ml ³ 1 ml (1%) w 10, 30 z z w (Table 2, 3). ƒƒ k 10 w k ƒ k z w ù kûš 20% k t ³ ³ w. Candida trpicalis 1% p +20% k cfu/ml z ƒ ù 1% +20% k z ƒ. Pichia anmala 1% p +20% k. ù 15% k z ƒ ùkû. k ƒ z w e w % k z ƒ š 20% k Table 1. Grwth inhibitin effect f yeasts with the varius sanitizers Pichia anmala Candida trpicalis Islate Y-1 Islate Y-2 Cntrl (N treatment) Ethanl(20%) Lactic acid(1%) Acetic acid(1%) Ethanl(20%) + Lactic acid(1%) ND 2) ND ND Ethanl(20%) + Acetic acid(1%) ND ND ND ND Ethanl(20%) + NaCl(10%) Parenthesis figures indicate the cntents f each sanitizers(v/v r w/v), 2) ND : Nt Detected. Each yeast strain cakes were added t each sanitizers fr 30 minutes and spreaded n PDA agar.

4 102 J.-S. Kim et al. Table 2. Grwth inhibitin effect f yeast after sanitatin with different ethanl cncentratins fr 10 minutes Candida trpicalis Pichia anmala Islate Y-1 Islate Y-2 ND : Nt Detected Cntrl(0%) 5% EtOH 10% EtOH 15% EtOH 20% EtOH 1% Acetic acid % Lactic acid ND 1% Acetic acid ND 1% Lactic acid % Acetic acid ND 1% Lactic acid % Acetic acid ND 1% Lactic acid ND Table 3. Grwth inhibitin effect f yeast after sanitatin with different ethanl cncentratins fr 30 minutes Candida trpicalis Pichia anmala Islate Y-1 Islate Y-2 ND : Nt Detected Cntrl(0%) 5% EtOH 10% EtOH 15% EtOH 20% EtOH 1% Acetic acid ND 1% Lactic acid ND ND 1% Acetic acid ND 1% Lactic acid ND 1% Acetic acid ND ND 1% Lactic acid ND 1% Acetic acid ND 1% Lactic acid x ƒ 30 w 15% k z ƒ x. w ¼ ¼ z w j ùkû. ù 10 z» w. y w k š w y w v w. ô Ð ò»ï ô Œ w k 10%» 1%, 3%, 5% Table 4. Grwth inhibitin effect f yeast fr different rganic acids f the cmplex sanitizer cntaining ten percentage ethanl Candida trpicalis Pichia anmala Islate Y-1 Islate Y-2 ND : Nt Detected 0% 1% 3% 5% Acetic acid ND ND Lactic acid ND Acetic acid ND ND Lactic acid Acetic acid ND ND Lactic acid Acetic acid ND ND Lactic acid ND ƒ w 100 ml ³ 1 ml (1%) w 30 z w w. k 10% w» z y wš w (Table 4).» ƒ z f ùkû» ƒ 3%ƒ z ƒ j ùkûš Pichia anmala wx š18) š. ù 5% w z š. k» ww z z w. ô Ð Ð»Ï ô Œ w 100 ml z 1 ml z ƒ ƒ 20 C 50 C 30 w z PDA w 48 z w (Table 5).» z ƒ 10 cfu/ml z 5-6 w. ù ³ Islate Y-1 Islate Y-2 w j š w p w w 30 w z ƒ. ù 20 C 50 C w

5 Grwth Inhibitin f Yeast Islated frm Prcessed Rice 103 Table 5. Grwth inhibitin effect f yeast fr different sanitizer temperature Pichia anmala Candida trpicalis Islate Y-1 Islate Y-2 Cntrl 20 C 50 C 20 C 50 C 20 C 50 C 20 C 50 C Ethanl(20%) + Acetic acid(1%) ND ND ND ND ND ND ND ND Ethanl(20%) +Lactic acid(1%) ND ND ND ND ND ND ND : Nt Detected w w z ƒ w ùkû.» z ƒ w x w z w. æ ƒì½ ä ô Ð Œ 20% k 1%, 1% p, w 30 ƒ w z w w ³ t ³ w wz s ƒw (Table 6). l œ w. ƒœt w z d w z w k» w z ³ z ƒ w y w ƒ. k ³ k ƒ ww w š ww w w ƒ w» ùkü». w š15) ³ 20 C, 5 ³ ƒ» k wz w. k 50% ƒ v w w» 1~2% w» 20% k 5 ü ³ z ƒ. ³ w k z šw Table 6. Detectin f yeast frm the stale prcessed rice cakes after varius sanitizers Viable cunt f yeast Cntrl Ethanl(20%) 2) Lactic acid(1%) Acetic acid(1%) Ethanl(20%) + Lactic acid(1%) Ethanl(20%) + Acetic acid(1%) ND 3) Ethanl(20%) + NaCl(10%) Nn-sanitizer treated, 2) Parenthesis figures indicate the cntents f each sanitizers(v/v r w/v), 3) ND : Nt Detected. The stale prcessed rice cakes were dipped in each sanitizers fr 30 minutes, stmached, and spreaded n PDA agar with erythrpmycin and chlramphenicl. ³ k y ww k 1 g w ³ y d w š17). k w w 50% ƒ ³ v w ù 8~10% ƒ w k 20% x w ³ ûš 10% ³ ƒ w. w ³ ù» s ƒœt» q» w z w. k ³ w z ƒ z k w w. ù» k» ƒ w z ³ k ƒœt qz w ƒ ƒ. w w hurdle technlgy multitarget preservatin (18) synergy z» w y w». ò w 2006 w w. ³ ƒœ ƒœt / y yw» w ƒœ t ƒ z w m wš w. q ƒœt qz w z Pichia anmala, Candida trpicalis k» w w w w. 20% k, 1% p, y 1% ƒƒ w z wx j ù k ƒƒ» yww z wƒ x w ƒ. 30 w z 6 lg cfu/ml¾ yw ƒ. w 20 C 50 C x w

6 104 J.-S. Kim et al. ³z ƒ f. 50 C 20% k 1% p y 30 w ƒœt» q³ z w w. w p w ³ m t z w t w y. Ù 1. Ghelardi, E., Celandrn, F., Salvetti, S., Barstti, C., Baggiani, A., and Senesi, S.: Identificatin and characterizatin f txigenic Bacillus cereus islate respnsible fr tw fdpsining utbreaks. FEMS Micrbil. Lett. 208, (2002). 2. Rice, K.M. and Piersn, M.D. Inhibitin f Salmnella by sdium nitrite and ptassium srbate in frankfurters. J. Fd Sci. 47, (1982). 3. Kim, D.J., Kwn, O.J., and Byun, M.W.: Cmbinatin effects f benzate, srbate and ph fr cntrl f Escherichia cli O157:H7. J. Fd Hyg. Safety 12, (1997). 4. Oh, D.H. and Marshall, D.L. Enhanced inhibitin f Listeria mncytgenes by glycerl mnlaurate with rganic acids. J. Fd Sci. 59, (1994). 5. Ita, P.S. and Hutkins, R.W.: Intacellular ph and survival f Listeria mncytgenes Sctt A in trypic sy brth cntaining acetic, lactic, citric, and hydrchlric acid. J. Fd Prt. 54, ( Ynng, K.M. and Fegding, P.M.: Acetic, lactic, citric acid and ph inhibitin f Listeria mncytgenes Sctt A and the effect n intracellular ph. J. Appl. Bacteril. 74, (1993). 7. Fernandes, C.F., Flick, G.J., Chen, J., and Thmas, T.B.: Rle f rganic acids during prcessing t imprve quality f channel catfish fillets. J. Fd Prt. 61, (1998). 8. Masuda, S., Hara-Kud, Y., and Kuagai, S.: Reductin Escherichia cli O157:H7 ppulatin in sy sauce, a fermented seasning. J. Fd Prt. 61, (1998). 9. Fischer, R., Fletcher, L., Cx, N.A., and Bailey, J.S.: Micrbilgical prperties f hard-cked eggs in a citric acid based preservative slutin. J. Fd Prt. 48, (1985). 10. Acuff, G.R., Vanderzant, C., Savell, J.W., Jnes, D.K., Griffin, D.B., and Ehlers, J.G.: Effect f acid decntaminatin f beef subprimal cuts n the micrbilgical and sensry characteristic f steaks. Meat Sci (1987). 11. Ahn, Y.S. and Shin, D.H.: Antimicrbial effects f rganic acids and ethanl n several fdbrne micrrganisms. Krean J. Fd Sci. Technl. 31, (1999). 12. Asplund, K., Nurmi, E., Hilli, P., and Hin, J.: The inhibitin f grwth f Bacillus cereus in liver sausage. Int. J. Fd. Mic. 7, (1988). 13. Jeng, J.H, Han, S.J, Ch, W.D., and Hwang, H.J.:Identificatin f spilage bacteria islated frm aseptic packaged cked rice and applicatin f acidic electrlyzed saline slutin as water-fr-cked rice. Krean J. Fd Sci. Technl. 31, (1999). 14. Shin, D.H, Kim, M.S., and Han, J.S.: Antimicrbial effect f ethanl extracts frm sme medicinal herbs and their fractinates against fd-brn bacteria. Krean J. Fd Sci. Technl. 29, (1997). 15. Jung, D.H.: Fdbrne Micrrganism Cntrl. pp ( Jang, J.H., Jang, J.S., Lee, S.Y., Kim, H.S., Kang, S.M., and Park, J.H.: Grwths inhibitin effects f ethanl and sdium chlride n Bacillus cereus. Krean J. Fd Sci. Technl. 35, (2003). 17. Lang, H.S., Weng, Y., and Rbin, Y.C.: Survial f Stapylcccus aureus and Escherichia cli as affected by ethanl and NaCl. J. Fd Prt. 64, ( Magnussn, J.: Brad and cmplex antifugal activity amng envirnmental islates f lactic acid bacteria. FEMS Micrbil. Lett. 14, (2003). 19. Leister, L.: Principle and applicatins f hurdle technlgy. In: GW Guld(ed.) New Methds f Fd Preservatin. Lndn. Blackie Academic & Prfessinal, pp (1995).

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