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Transkript:

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ÖZET Yüksek Lisans Tezi 67$57(5.h/7h5.8//$1,/$5$.h5(7ø/(1+,<$5785ù8/$5,1'$ %ø<2-(1$0ø12/8ù808h=(5ø1($5$ù7,50$ Zeynep TURGUT Ankara Üniversitesi Fen Bilimleri Enstitüsü *ÕGD0 KHQGLVOL L$QDELOLP'DOÕ 'DQÕúPDQ3URI'U.DPXUDQ AYHAN %X odoõúpdgd KHP GR DO IHUPDQWDV\RQ \ROX\OD KHPGHLactobacillus plantarum 11B VWDUWHU N OW U NXOODQÕODUDN KÕ\DU WXUúXVX UHWLPL \DSÕOPÕú YH IHUPDQWDV\RQ V UHVLQFH ROXúDQEL\RMHQDPLQOHULQPLNWDUODUÕ< NVHN3HUIRUPDQVOÕ6ÕYÕ.URPRWRJUDILVL+3/& \ QWHPL LOH EHOLUOHQPLúWLU $\UÕFD EHOLUOL J QOHUGH DOÕQDQ UQHNOHUGH WRSODP PH]RILOLN aerobik bakteriler (TMAB), laktik asit bakterileri (LAB), enterobakteri ve koliform grubu bakteriler ile maya-n I VD\ÕPODUÕ YH S+ WLWUDV\RQ DVLWOL L YH WX] JLEL NLP\DVal DQDOL]OHUGH\DSÕOPÕúWÕU 6WDUWHUNDWÕODUDN UHWLOHQ KÕ\DUWXUúXVX UQHNOHULQGHS+WLWUDV\RQ DVLWOL L WX]NRQWURO UQHNOHULQGHS+WLWUDV\RQDVLWOL LYHWX]RODUDN EXOXQPXúWXU)HUPDQWDV\RQV UHVLQFHHQ\ NVHN/$%70$%YHPD\DVD\ÕODUÕVWDUWHUOL UQHNOHUGH VÕUDVÕ\OD YH log kob/ml NRQWURO UQHNOHULQGHVÕUDVÕ\OD 8,35, 8,32 ve 5,56 log kob/ml RODUDN EHOLUOHQPLúWLU Küf, enterobakteri ve koliform VD\ÕP VRQXoODUÕ ORJ NREP/ GHQ G ú N EXOXQPXúWXU 'HQHPHGH NXOODQÕODQ Lactobacillus plantarum % QLQ 056 VÕYÕ EHVL\HULQGH ROXúWXUGX X WRSODP EL\RMHQ DPLQ PLNWDUÕQÕQ JP/ ROGX X EHOLUOHQPLúWLU)HUPDQWDV\RQXQ ELWLPLQGH VWDUWHU NDWÕODQ UQHNOHUGH VÕUDVÕ\OD SXWUHVLQ JP/-salamura), feniletilamin (4,633 µg/ml-salamura) ve triptamin (1,936 µg/mg-meyve); kontrol örneklerinde ise feniletilamin (29,817 µg/ml-salamura), putresin (20,492 µg/mg-meyve) ve triptamin (9,246 µg/ml-salamura) en yüksek konsantrasyona sahip biyojen aminler olarak belluohqplúwlu 6SHUPLQ YH KLVWDPLQ G ]H\OHULQLQ LVH GR DO IHUPDQWDV\RQOD UHWLOHQ NRQWURO UQHNOHULQGH VÕUDVÕ\OD JP/-salamura ve 3,810 µg/mg-meyve) starter NDWÕODUDN UHWLOHQ KÕ\DU WXUúXODUÕQD NÕ\DVOD GDKD \ NVHN ROGX X VDSWDQPÕúWÕU.ontrol örnekleulqghnl IHQLOHWLODPLQ PLNWDUODUÕKDULoIHQLOHWLODPLQ SXWUHVLQKLVWDPLQ WLUDPLQ YHWRSODPEL\RMHQDPLQPLNWDUODUÕQÕQWRNVLNOLPLWOHULQDOWÕQGDROGX XDQODúÕOPÕúWÕU 2006, 72 sayfa Anahtar Kelimeler:7XUúXEL\RMHQDPLQIHUPDQWDV\RQ+3/&ODNWLNDVLW bakterileri i

ABSTRACT Master Thesis A RESEARCH ON FORMATION OF BIOGENIC AMINE IN CUCUMBER PICKLES PRODUCED BY USING STARTER CULTURE Zeynep TURGUT Ankara University Graduate School of Natural and Applied Sciences Department of Food Engineering Supervisor: Prof. Dr. Kamuran AYHAN In this study, cucumber pickle was produced by spontaneous fermentation or using Lactobacillus plantarum 11B as starter culture, then amount of biogenic amines, which were produced during fermentation, was determined by High Performance Liquid Chromatography (HPLC). Additionally, on certain days during fermentation, enumeration of total mesophilic aerobic bacteria (TMAB), lactic acid bacteria (LAB), enterobacteria, coliforms and yeast-mold were performed, as well as the chemical analyses such as ph, titration acidity and salt. The ph, titration acidity and salt content of the samples produced by adding starter culture were found as 3.43, 0.88% and 4.20%, respectively, while those of control samples were 3.45, 0.98% and 4.15%, respectively. The highest LAB, TMAB and yeast counts during fermentation in starter added samples were 8.58, 8.55 and 4.75 log cfu/ml, respectively; in control samples those were 8.35, 8.32 and 5.56 log cfu/ml, respectively. Mold, enterobacteria and coliform counts were found lower than 1 log cfu/ml. Lactobacillus plantarum 11B strain, which was used as starter culture for the experiments, produced 0.413 µg/ml total biogenic amine in MRS broth. At the end of the fermentation period, putrescine (11.863 µg/ml-brine), phenylethylamine (4.633 µg/ml-brine) and tryptamine (1.936 µg/mg-fruit) were determined as the biogenic amines which had the highest concentration in starter added samples. Whereas phenylethylamine (29.817 µg/ml-brine), putrescine (20.492 µg/mg-fruit) and tryptamine (9.246 µg/ml-brine) displayed the highest concentrations in control samples. On the other hand, spermine and histamine levels were higher in control samples (6.536 µg/ml-brine and 3.810 µg/mg-fruit, respectively) produced by spontaneous fermentation than the samples produced by adding starter culture. It was determined that phenylethylamine, putrescine, histamine tyramine and total biogenic amine contents of the samples were lower than the toxic limits, except the phenylethylamine content of control samples. 2006, 72 pages Key Words: Pickle, biogenic amine, fermentation, HPLC, lactic acid bacteria ii

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dø=(/*(/(5'ø=ø1ø dl]hojh6wduwhundwnõoõ UQHNOHUHDLWDQDOL]SODQÕ 19 Çizelge 3.2 Kontrol örnekleulqhdlwdqdol]sodqõ 19 dl]hojh+3/&flkd]õqõq ]HOOLNOHULYHEL\RMHQDPLQDQDOL]LLoLQ NURPDWRJUDILNRúXOODUÕ.. 21 Çizelge 4.1 Starter kültür adaptasyonu... 26 dl]hojh.rqwuroyhvwduwhun OW UNDWÕODUDN UHWLOHQKÕ\DU WXUúXVXQGDNLNLP\DVDOGH LúLPOHU... 27 dl]hojh.rqwuroyhvwduwhun OW UNDWÕODUDN UHWLOHQKÕ\DU WXUúXVXQGDNLPLNUREL\RORMLNGH LúLPOHU 31 dl]hojh9hulohulqgr UXVDOOÕ Õ... 39 dl]hojh.rqwuroyhvwduwhun OW UNDWÕODUDN UHWLOHQKÕ\DU WXUúXODUÕQGDNLWULSWDPLQPLNWDUODUÕ 40 Çizelge 4.6 Kontrol ve swduwhun OW UNDWÕODUDN UHWLOHQKÕ\DU WXUúXODUÕQGDNLIHQLOHWLODPLQPLNWDUODUÕ 43 dl]hojh.rqwuroyhvwduwhun OW UNDWÕODUDN UHWLOHQKÕ\DU WXUúXODUÕQGDNLSXWUHVLQPLNWDUODUÕ... 46 dl]hojh.rqwuroyhvwduwhun OW UNDWÕODUDN UHWLOHQKÕ\DU WXUúXODUÕQGDNLNDGDYHULQPLNWDUODUÕ 48 Çizelge 4.9 Kontrol ve starter N OW UNDWÕODUDN UHWLOHQKÕ\DU WXUúXODUÕQGDNLKLVWDPLQPLNWDUODUÕ 51 dl]hojh.rqwuroyhvwduwhun OW UNDWÕODUDN UHWLOHQKÕ\DU WXUúXODUÕQGDNLWLUDPLQPLNWDUODUÕ... 53 dl]hojh.rqwuroyhvwduwhun OW UNDWÕODUDN UHWLOHQKÕ\DU WXUúXODUÕQGDNLVSHUPLGLQPLNWDUODUÕ56 dl]hojh.rqwuroyhvwduwhun OW UNDWÕODUDN UHWLOHQKÕ\DU WXUúXODUÕQGDNLVSHUPLQPLNWDUODUÕ 58 dl]hojh.rqwuroyhvwduwhun OW UNDWÕODUDN UHWLOHQKÕ\DU WXUúXODUÕQGDNLWRSODPEL\RMHQDPLQPLNWDUODUÕ... 61 vii

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.$<1$.g=(7/(5ø %L\RMHQ DPLQOHU JÕGDODUGD ED]Õ VSHVLILN DPLQR asitlerin dekarboksilasyonuna veya DOGHKLWYHNHWRQODUÕQWUDQVDPLQDV\RQXQDED OÕRODUDNROXúDQWHPHOD]RWOXELOHúLNOHUGLU Histidin, tirozin, triptofan ve fenilalaninden mono aminler olan histamin (HIS), tiramin (TRA), triptamin (TRP) ve feniletilamin (FE$ PH\GDQD JHOLUNHQ EHQ]HU úhnlogh RUQLWLQ YH OLVLQGHQ GLDPLQOHU RODQ SXWUHVLQ 387 YH NDGDYHULQ.$' ROXúPDNWDGÕU Poliaminlerden spermidin (SPD) ve spermin (SPM) ise putresinden meydana JHOPHNWHGLUùHNLO+DOiV]et al. 1994, Moret and Conte 1996,.DODþ et al. 2000a,b,.DODþ Anonymous 2006). ' ú N PROHN OHU D ÕUOÕ D VDKLS RODQ EL\RMHQ DPLQOHU GR DO EL\RORMLN DNWLYLWHOHUL QHGHQL\OHSHNoRNJÕGDGDEXOXQPDNWDGÕUODU%XJÕGDODUDUDVÕQGDEDOÕNYHEDOÕN U QOHUL et ve et ürünleri (Maijala et al. 1995a, Bodmer et al. 1999), yumurta, peynir gibi SURWHLQFH]HQJLQJÕGDODUIHUPHQWHVHE]HOHUYHVR\DOÕ U QOHUELUDYHúDUDSJLELDONROO LoHFHNOHU 6KDODE\.tåHN DQG 3HOLNiQRYi PH\YHOHU IÕQGÕN YH olnrodwd 6LOOD6DQWRVJLELJÕGDODU\HUDOPDNWDGÕU *ÕGDODUGD EL\RMHQ DPLQ ROXúXPX KHP GHNDUERNVLOD] HQ]LP UHWPH \HWHQH L RODQ PLNURRUJDQL]PDODUÕQ YDUOÕ ÕQD KHP GH X\JXQ NRIDNW U YHYH\D ELU LQG NOH\LFL LOH GHNDUERNVLOD] HQ]LP VHQWH]L LoLQ NRúXOODUÕQ X\JXQ ROPDVÕQD YH RUWDPGDNL DPLQR DVLt NRQVDQWUDV\RQXQD ED OÕGÕU %X QHGHQOH \ NVHN PLNWDUGD EL\RMHQ DPLQ ROXúXPXQX NRQWURO DOWÕQD DODELOPHN LoLQ ROXúXPX HWNLOH\HQ NULWLN IDNW UOHULQ PXWODND ELOLQPHVL gerekmektedir (Maijala et al. 1995b). 3

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$PLQR DVLW GHNDUERNVLOD] HQ]LPL EDNWHULOHU DUDVÕQGD \D\JÕQ ROPDPDNOD ELUOLNWH Achromobacter, Bacillus, Citrobacter, Clostridium, Escherichia, Klebsiella, Lactobacillus, Pediococcus, Photobacterium, Proteus, Pseudomonas, Salmonella, Shigella, Streptococcus ve Vibrio cinsi pek çok bakteri bir veya daha fazla amino asidi GHNDUERNVLOH HWPH \HWHQH LQGHGLU 6XPQHU DQG 7D\ORU WHQ %ULQN et al. 1990, Hwang et al. 1995, Silla Santos 1996). Birçok Enterobacteriaceae türü putresin ve/veya kaddyhulq UHWPH \HWHQH LQH VDKLSNHQ \DOQÕ]FD ELUNDo Lactobacillus VXúXQXQ KLVWLGLQ dekarboksilaz enzimi pozitiftir (ten Brink et al. 1990). Ayhan et al. DGHWNÕ\PD UQH LQGHQL]ROHHGLSWDQÕPODGÕNODUÕWRSODPDGHW (QWHUREDFWHULDFHDH VXúXQX 7ULStik Soy Broth besiyerinde üreterek ml deki PLNURRUJDQL]PD VD\ÕODUÕ LOH EHVL\HULQH NDWWÕNODUÕ KLVWLGLQLQ EDNWHULOHU WDUDIÕQGDQ NXOODQÕOPDVÕ VRQXFX ROXúDQ KLVWDPLQ G ]H\OHULQL +3/& WHNQL L LOH EHOLUOHPLúOHUGLU 'L HU \DQGDQ VXúODUÕQ PRGLIL\H 1LYHQ $JDU EHVLyerinde histidin dekarboksilaz enzim DNWLYLWHOHULQL GH LQFHOHPLúOHUGLU 6RQXo RODUDN PLNURRUJDQL]PD VD\ÕVÕ LOH UHWLOHQ KLVWDPLQG ]H\LDUDVÕQGDELUNRUHODV\RQVDSWD\DPDGÕNODUÕQÕHafnia alvei1vxúxqxq EHVL\HULQGHNL VD\ÕVÕQÕQ ORJ NREP/ G ]H\LQH XODúWÕ Õ KDOGH KLVWLGLQL HQ D] LQGLUJH\HQ VXú ROGX XQX LIDGH HWPLúOHUGLU 0RGLIL\H 1LYHQ RUWDPÕQÕQ KLVWLGLQ dekarboksilaz enzim üretiminin EHOLUOHQPHVLQGH KDWDOÕ SR]LWLI VRQXoODU ROXúWXUDELOGL L VRQXFXQDYDUPÕúODUGÕU Durlu et al. NÕ\PD YH KDPEXUJHUOHUGHQ L]ROH HGLOLS WDQÕPODQPÕú EnterobacteriaceaeIDPLO\DVÕQDDLWDGHWVXúXQ%UDLQ+HDUW,QIXVLRQVÕYÕEHVL\HULQGH EL\RMHQDPLQ UHWPH \HWHQHNOHULQLDUDúWÕUGÕNODUÕoDOÕúPDODUÕQGDE. coli (& VXúXQXQ 65,88 mg/100 ml ile en yüksek düzeyde histamin üretwl LQL DUGÕQGDQ E. coli EC03 VXúXQXQPJP/G ]H\LQGHNDGDYHULQYHCitrobacter freundii CF02 nin ise PJ P/ SXWUHVLQ UHWLFLVL ROGX XQX VDSWDPÕúODUGÕU (QWHUREDFWHULDFHDH IDPLO\DVÕQD PHQVXS VXúODUÕQ UHWWLNOHUL HVDV EL\RMHQ DPLQOHULQ KLstamin, putresin ve NDGDYHULQROGX XQXLOHULV UP úohuglu 5

%LOLQGL L JLEL VWDUWHU N OW U NXOODQÕPÕ NÕVD V UHGH VWDQGDUW NDOLWHGH ELU JÕGD UHWLPLQL RODQDNOÕ NÕOPDNWDGÕU %X QHGHQOH \DSÕODQ DUDúWÕUPDODUGD JÕGD IHUPDQWDV\RQODUÕQGD NXOODQÕODQ VWDUWHU N OW Ulerin seçiminde sadece laktik asit üretiminin göz önünde WXWXOPDPDVÕ EL\RMHQ DPLQ ROXúWXUPD \HWHQHNOHULQLQ GH ELOLQPHVLQLQ JHUHNOL ROGX X YXUJXODQPDNWDGÕU6WUDXEet al. 1995, Halász et al. 1999). )DHWK YH 5RGOHU KHP ohúlwol JÕGD PDGGHOHULQGHQ L]Rle ettikleri hem de kendi HQVWLW OHULQGHNLW PODNWLNDVLWEDNWHULOHULQLQVÕYÕN OW UGHROXúWXUGXNODUÕEL\RMHQDPLQ G ]H\OHULQL +3/& WHNQL L NXOODQDUDN LQFHOHPLúOHU YH or X VXúXQ G ú N G ]H\GH KLVWDPLQ UHWWL LQL DQFDN Lactobacillus buchneri B189 ve B190 QXPDUDOÕ VXúODUÕQÕQ 49,7-41,3 mg/l gibi yüksek düzeyde histamin, yine Streptococcus B218 in de 408 PJ/G ]H\LQGHWLUDPLQROXúWXUGX XQXEHOLUOHPLúOHUGLU Straub et al. ohúlwol JÕGDODUGDQ L]ROH HWWLNOHUL DGHW ODNWLN DVLW EDNWHULVLQLQ biyojen amlq ROXúWXUPD \HWHQH LQL DUDúWÕUPÕúODUGÕU ddoõúpdgd L]ROH HGLOHQ VXúODUÕQ SXWUHVLQNDGDYHULQKLVWDPLQWLUDPLQYHIHQLOHWLODPLQROXúWXUPD\HWHQHNOHULQHEDNÕOPÕú ve Carnobacteria spp., Lactobacillus buchneri, L. curvatus ve L. reiteri nin yüksek düzeyde EL\RMHQ DPLQ ROXúWXUGX XQX EHOLUOHPLúOHUGLU 6 W U QOHUL úduds YH ODKDQD WXUúXVXQGD \D\JÕQ RODUDN NXOODQÕODQ ED]Õ VWDUWHU N OW UOHULQ \ NVHN G ]H\GH EL\RMHQ DPLQ ROXúWXUGX XQX EX QHGHQOH VWDUWHU N OW U VHoLPLQGH EX ]HOOLNOHULQ GLNNDWH DOÕQPDVÕJHUHNWL LQLYXUJXODPÕúODUGÕU Aytaç vd. (1997), Morganella morganii ve Proteus vulgarislohklvwdplqroxúxpxqdhwnl HGHQED]ÕSDUDPHWUHOHULLQFHOHGLNOHULoDOÕúPDODUÕQGD UQHNOHUGHNLKLVWDPLQPLNWDUODUÕQÕ NDORULPHWULN \ QWHPOH VSHNWURIRWRPHWULN RODUDN EHOLUOHPLúlerdir. Buna göre; Tripton 6R\D+LVWLGLQ76+VÕYÕEHVL\HULQGHM. morganii nin optimum ph 5,0 ve % 4 NaCl konsantrasyonunda, P. vulgaris in ise optimum ph 6,0 ve % 1 NaCl konsantrasyonunda maksimum düzeyde histamin ürettiklerini; her iki bakterinin de 30 o C da ve 24 saatte en yüksek histamin üretimi (M. morganii için 291,21 µg/ml, P. vulgarislolq JP/VHYL\HVLQHXODúWÕNODUÕQÕEXOPXúODUGÕU 6

%L\RMHQ DPLQOHU Y FXWWD SURWHLQ KRUPRQ YH Q NOHLN DVLW VHQWH]LQLQ LON EDVDPD ÕQÕ ROXúWXUXUODU (Hornero-Méndez and Garrido-Fernández 1997). Putresin, spermin, VSHUPLGLQ JLEL SROLDPLQOHU FDQOÕ K FUHOHULQ YD]JHoLOPH] ELOHúHQOHULGLU $\UÕFD SROLDPLQOHUED ÕUVDNODUGDNLLPP QRORMLNVLVWHPGHYHQRUPDOPHWDEROLNIRQNVL\RQODUÕQ aktivitesinin sürdürülmesinde de gerekolglu6lood6dqwrv*õgdodugdnlsxwuhvlqyh SROLDPLQOHULQ VÕQÕUOÕ PLNWDUODUGD DOÕQPDVÕ LQVDQ EHVOHQPHVLQGH ROXPOX ELU URO R\QDPDVÕQD NDUúÕQ ]HOOLNOH KLVWDPLQ YH WLUDPLQ JLEL EL\RMHQ DPLQOHULQ D Õ] \ROX\OD DúÕUÕPLNWDUODUGD DOÕQPDVÕ VRQXFXQGD PLGH EXODQWÕVÕ NXVPD DúÕUÕ \RUJXQOXN VROXQXP J oo NDOS odusõqwõvõ EDú D UÕVÕ EDú G QPHVL KLSRWDQVL\RQ KLVWDPLQ SXWUHVLQ NDGDYHULQ YH\D KLSHUWDQVL\RQ WLUDPLQ NXUGHúHQ EH\LQ NDQDPDVÕ DQDILODNWLN úrn sendromu ve ölüm meydana gelmektedir (Moret et al..tåhn DQG 3HOLNiQRYi 1998, Lange et al. 2002, Erkmen and Bozkurt 2004, McCabe-Sellers et al. 2006). Memelilerin intestinal bölgelerinde oldukça etkili detoksifikasyon sistemleri mevcuttur. Bu detoksifikasyon sisteminde mono amin oksidaz (MAO) ve di amin oksidaz (DAO) HQ]LPOHUL QHPOL URO R\QDPDNWDGÕU WHQ %ULQN et al. âslþnd et al. 2002). 'HWRNVLILNDV\RQXQ HWNLQOL L ELUH\OHU DUDVÕQGD IDUNOÕOÕN J VWHUHELOGL L JLEL ED]Õ antidepresan ilaçlar ve alkol gibi MAO-LQKLELW UOHULQLQ Y FXGD DOÕPÕQGan da HWNLOHQPHNWHGLU 6WUHV GHSUHV\RQ YH 3DUNLQVRQKDVWDOÕ ÕQÕQ WHGDYLVLQGH NXOODQÕODQ D UÕ NHVLFLYHGL HULODoODU0$2-LQKLELW UOHULLoHUPHNWHGLU.DODþ et al. 2000a,b, Moret et al. 2005, McCabe-Sellers et al. 7LUDPLQ YH\D GL HU DPLQOHUL LoHUHQ ROJXQODúPÕú SH\QLU úduds ELUD YH PD\D HNVWUDNWODUÕ\OD ELUOLNWH VDXHUNUDXW VXVX] ODKDQD WXUúXVX bakla ve avokado gibi sebze ve meyvelerin tüketimini takiben hipertansiyon krizlerinin J U OG YHVDDWOLN]DPDQGLOLPLLoLQGHWLUDPLQDOÕPÕQÕQPJ ÕJHçmesinin hasta LoLQWHKOLNHROXúWXUGX XELOGLULOPHNWHGLU0RUHWet al. 2005). %L\RMHQ DPLQOHULQ ]DUDUOÕ HWNLOHUL NLúLOHULQ IL]\RORMLN \DSÕVÕQD WROHUH HGHELOGLNOHUL konsantrasyona, ortamdaki biyojen aminlerin birbirleri ile interaksiyonuna ve MAO enzimlerlqlq DNWLYLWHVLQH ED OÕ RODUDN GH LúPHNWHGLU WHQ %ULQN et al. 1990). Feniletilamin, putresin ve kadaverin histamin detoksifikasyon enzimleri olan DAO ve HMT (histamin N-metiltransferaz) enzimlerinin her ikisini de inhibe ederken; tiramin 0$2 ODUÕWULSWDPLQGH'$2 ODUÕLQKLEHHWPHNWHGLU6WUDWWRQet al. 1991). 7

%L\RMHQ DPLQOHUGHQ ND\QDNODQDQ JÕGD ND\QDNOÕ LQWRNVLNDV\RQODU LoLQGH HQ ID]OD UDVWODQDQ KLVWDPLQR]LVWLU %LUH\OHU DUDVÕQGD GHWRNVLILNDV\RQ PHNDQL]PDVÕ IDUNOÕOÕN J VWHUPHNOH ELUOLNWH JÕGDODUGD HQ \ ksek toksik limit 100 mg histamin/kg, alkollü LoHFHNOHUGH LVH PJ/ RODUDN UDSRU HGLOPLúWLU WHQ %ULQN et al. 'L HU EL\RMHQ DPLQOHULQ WRNVLN GR]ODUÕ KDNNÕQGD GDKD D] YHUL EXOXQPDNOD ELUOLNWH HúLN GH HUOHULQLQ tiramin için 100-800 mg/kg, feniletilamin için 30 mg/kg ve toplam biyojen amin için 100-200 mg/kg ROGX XELOGLULOPLúWLU0DLMDODDQG(RUROD Nout 1994, Halász et al. 1999). $\UÕFD 6WUDWWRQ et al WDUDIÕQGDQ SXWUHVLQ LoLQ EHOLUOHQHQ WRNVLN OLPLWLQ PJNJROGX XLOHULV U OP úw U *ÕGDODUGD EL\RMHQ DPLQOHULQ EXOXQXúX YH DUWÕúÕQGDQ NDoÕQÕOPDVÕQÕQ ELU QHGHQL GH SXWUHVLQ VSHUPLQ YH VSHUPLGLQ JLEL VHNRQGHU DPLQOHULQ QLWULW LOH ELUOHúHUHN NDQVHURMHQ QLWUR]DPLQOHUHG Q úphohulgluwhq%ulqnet al. 1990). Ülkemiz de dahil olmak üzere phnorn ONHGHIHUPHQWHJÕGDODUGDEL\RMHQDPLQROXúXPX YHPLNWDUODUÕ ]HULQHoRNVD\ÕGDDUDúWÕUPD\DSÕOPÕúWÕUg]HOOLNOHIHUPHQWHHW U QOHULQGH (Maijala and Eerola 1993, Maijala et al. 1995a,b, Cantoni 1995, Hernandez-Jover et al. 1997a,b, Ayhan et al. 1999, Aytac et al. 2000, Komprda et al. 2001, Durlu-Özkaya et al. 2001, Cosansu and Ayhan 2002, Ansorena et al. 2002, Gonzalez-Fernandez et al. 2003, Bruna et al. 2003, Komprda et al. 2004, Erkmen and Bozkurt 2004, Bozkurt YH ohúlwol SH\QLUOHUGH 6FKQHOler et al. 1997, Durlu-Özkaya et al. 1999, Durlu- Özkaya et al. 2000, Durlu-Özkaya 2001, Durlu-Özkaya 2002, Novella-Rodriguez et al. 2002, Gennaro et al. 2003, Novella-Rodriguez et al. 2003, Sagun et al. 2005) biyojen DPLQOHULQROXúXPXYHPLNWDUÕQÕQEHOLUOHQPHVL ]HULQHSHNoRNoDOÕúPDEXOXQPDNWDGÕU <DSÕODQ DUDúWÕUPDODUGD (OHNWURNLQHWLN.DSLOOHU (OHNWURIRUH] %LRVHQV U YH +3/& JLEL ohúlwol\ QWHPOHUNXOODQÕOPÕúWÕU )HUPHQWH U QOHULoLQGH\HUDODQYH\D\JÕQRODUDNW NHWLOHQWXUúXODUÕQ UHWLPV UHFLQGH biyomhq DPLQ ROXúXPXQGD \LQH KDPPDGGH VÕFDNOÕN S+ IHUPDQWDV\RQ V UHVL GR DO IORUDYH\DNDWÕODQVWDUWHUN OW UJLELSHNoRNIDNW U QUROR\QDGÕ ÕDoÕNWÕU 8

GH.HLO YH.ULWWHU VDXHUNUDXWGD VXVX] ODKDQD WXUúXVX SXWUHVLQ YH KLVWDPLQ EXOXQGX XQX EHOLUOHPLúOHUGLU $\QÕ DUDúWÕUPDFÕODU ODKDQDGD VHUEHVW KLVWLGLQ LoHUL LQL LQFHOHPLúOHU DQFDN YDUOÕ ÕQÕ VDSWD\DPDPÕúODUGÕU 6RQXo RODUDN histaminin histidin DPLQR DVLGLQLQ HQ]LPDWLN UHDNVL\RQD X UDPDVÕ VRQXFX PH\GDQD JHOGL LQL LOHUL V UP úohuglu+doiv]et al. 1999). Mayer et al. (1973), 50 adet WLFDULVDXHUNUDXW UQH LQGHNLKLVWDPLQNRQVDQWUDV\RQODUÕQÕ LQFHOHPLúOHUYHEXNRQVDQWUDV\RQODUÕQ-PJNJDUDVÕQGDGH LúWL LQLEHOLUOHPLúOHUGLU 'L HU ELU odoõúpd %XFNHQK VNHV et al. WDUDIÕQGDQ \DSÕOPÕú YH DUDúWÕUÕFÕODU sauerkraut UQHNOHULQGH KLVWDPLQ NRQVDQWUDV\RQODUÕQÕ -78 mg/kg olarak VDSWDPÕúODUGÕU%DUiWKYH+DOiV]DGHWsauerkraut UQH LQGHKHPVHE]HKHPGH VDODPXUDGDKLVWDPLQG ]H\OHULQLDUDúWÕUPÕúODUGÕU+LVWDPLQNRQVDQWUDV\RQXQXQODKDQDGD 42-52 mg/kg, salamurada ise daha yüksek konsantrasyonda (143-PJNJROGX XQX EHOLUOHPLúOHUGLU+DOiV]et al. 1999). Taylor et al. ohúlwol PDUNHWOHUGH VDWÕúD VXQXODQ DGHW VDXHUNUDXW UQH LQLQ KLVWDPLQ LoHUL LQL DUDúWÕUPÕúODU YH -13,0 mg/100 g olarak belirledikleri histamin GH HUOHULQLQJÕGD]HKLUOHQPHOHULQHQHGHQRODQPJNJVHYL\HVLQGHQG ú NROGX X VRQXFXQDYDUPÕúODUGÕU(NLFLDQG&RVNXQ Kuensch et al.wlfdulrodudnvdwõúdvxqxodqsauerkraut UQHNOHULLOH\DSWÕNODUÕELU çdoõúpdgd LQFHOHQHQ UQHNOHULQ WDPDPÕQÕQ QHPOL Oo GH HQ \ NVHN PJNJ EL\RMHQDPLQLoHUGL LQLVDSWDPÕúODUGÕU Kuensch et al. VDXHUNUDXW IHUPDQWDV\RQXQXQ EDúODQJÕo DúDPDVÕQGD SXWUHVLQ PLNWDUÕQÕQ PJNJ GDQ GDKD ID]OD ROGX XQX YH IHUPDQWDV\RQXQ VRQXQD GR UX KLVWDPLQ YH WLUDPLQ DPLQOHULQLQ J U OG Q ELOGLUPLúOHUGLU +RUQHUR-Méndez and Garrido-Fernández 1997). Halász et al. (1999) starter kültür seçiminin sauerkraut fermantasyonu üzerine etkisini DUDúWÕUGÕNODUÕ odoõúpdoduõqgd LNL IDUNOÕ Vtarter kültürün biyojen amin üretim düzeyini 9

LQFHOHPLúOHUGLU 056 VÕYÕ EHVL\HULQGH Lactobacillus curvatus 2775 in histamin VHQWH]OHPHGL LQLYH UHWWL LWRSODPDPLQGH HULQLQGHG ú NoÕNWÕ ÕQÕEHOLUOHPLúOHUGLU 6WDUWHU N OW U NXOODQÕPÕQÕQ ODKDQD IHUPDQWDVyonu süresince ph ve biyojen amin ROXúXPXQXQ D]DOPDVÕQÕ ROXPOX \ QGH HWNLOHGL LQL L. plantarum XQ G ú N G ]H\GH DPLQ UHWLFLVL ROGX XQX YH EHOLUOHQHELOLU PLNWDUGD KLVWLGLQ GHNDUERNVLOD] HQ]LP DNWLYLWHVLQHVDKLSROGX XQXELOGLUPLúOHUGLU%XQDJ UHVWDUWHU kültür olarak L. plantarum VHoLOGL LQGHJODKDQDLoLQ 6-2.10 7 G ]H\LQGHNXOODQÕOPDVÕQÕDQFDNL. curvatus LoLQ WLUDPLQ NRQVDQWUDV\RQXQX DUWÕUPDPDNDPDFÕ\ODGDKD G ú N G ]H\GH NXOODQÕOPDVÕ JHUHNWL LQLYXUJXODPÕúODUGÕU.DODþ et al. (1999) 121 adet VDXHUNUDXW UQH LQGH K FUHVHO HOHNWURNLQHWLN NDSLOHU NURPRWRJUDIL \ QWHPL LOH EL\RMHQ DPLQOHUL EHOLUOHPLúOHUGLU guqhnohu dhn UHWLPL $YXVWXU\D UHWLPL HY \DSÕPÕ YH VDODPXUDVÕ\OD ELUOLNWH NDYDQR]GD VWHULOL]H HGLOHQOHU ROPDN ]HUH G UW JUXS DOWÕQGD WRSODQPÕúWÕU 7LUDPLQ SXWUHVLQ YH NDGDYHULQ LoLQ HQ \ NVHN NRQVDQWUDV\RQODU VÕUDVÕ\OD YH PJNJ RODUDN EHOLUOHQPLúWLU (Y \DSÕPÕ VDXHUNUDXWODUGD LVH orn G ú N NRQVDQWUDV\RQODU EHOLUOHPLúOHUGLU +LVWDPLQ VHYL\HVLQLQ UQHNOHULQ QGH PJNJ ÕQ DOWÕQGD YH UQHNOHULQ \DOQÕ]FD XQXQPJNJ ÕQ ]HULQGHROGX XQXEXQDNDUúÕQWULSWDPLQ(1,3 mg/kg) ve spermidinin PJNJHQG ú NNRQVDQWUDV\RQDVDKLSEL\RMHQDPLQOHUROGX XQXVDSWDPÕúODUGÕU.DODþ et al. D DGHW EH\D] ODKDQD ohúlglql NXOODQDUDN IDUNOÕ VDXHUNUDXW KD]ÕUODPÕúODUGÕU%DúODQJÕoWD o & GDJ QIHUPDQWDV\RQDEÕUDNÕODQYHGDKDVRQUD 5-6 o C da depolanan UQHNOHUL D\ VRQXQGD DQDOL] HWPLúOHUGLU $UDúWÕUPDGD VWDUWHU NDWÕOPDGDQ IHUPDQWDV\RQD X UDWÕODQ NRQWURO UQHNOHUi ile Lactobacillus plantarum, Lactobacillus casei, Pediococcus pentosaceus, Enterococcus faecium YH KD]ÕUODQDQ NDUÕúÕN0LFURVLOWLFDULODNWLNDVLWEDNWHULW UOHULKHUELULQGHNLGR] 6 kob/g lahana) ile LQRNXOHHGLOHQ UQHNOHUROPDN ]HUHIDUNOÕGHQHPHNXUXOPXúYH UQHNOHUGHNLEL\RMHQ DPLQ ROXúXPX K FUHVHO HOHNWURNLQHWLN NDSLOHU NURPDWRJUDIL \ QWHPL\OH LQFHOHQPLúWLU $QDOL] VRQXFXQGD KLVWDPLQ WULSWDPLQ VSHUPLGLQ YH VSHUPLQ NRQVDQWUDV\RQODUÕQÕ JHQHOOLNOH PJNJ ÕQ DOWÕQGD EXOPXúODUGÕU 7LUDPLQ SXtresin ve kadaverin ROXúXPXQXQ WRSODP NRQVDQWUDV\RQXQX NRQWURO UQHNOHULQGH -780 mg/kg, L. plantarum NXOODQÕODQ UQHNOHUGH - PJNJ 0LFURVLO NXOODQÕODQ UQHNOHUGH LVH 10

41,1- PJNJ RODUDN EHOLUOHPLúOHUGLU L. plantarum ve Microsil in bu oranlduõq G ú ú QGH QHPOLUROR\QDGÕNODUÕQÕLVWDWLVWLNVHORODUDNGDVDSWDPÕúODUGÕU.DODþet al. E \DSWÕNODUÕGL HU ELU odoõúpdgd EH\D] ODKDQD ohúlgl LOH UHWLOPLú U QOHULQLQ GHSRODQPDVÕ V UHFLQGH EL\RMHQ DPLQ ROXúXPX YH DPLQ NRQVDQWUDV\RQODUÕQGDNL GH LúLPOHUL LQFHOHPLúOHUGLU 'HQHPHGH WXUúXODUÕ o C da 14 J QV UH\OHGR DOIHUPDQWDV\RQDEÕUDNWÕNWDQVRQUD-6 o & GDGHSRODPÕúODUYHYH D\ÕQ VRQXQGD UQHN DOPÕúODUGÕU $QDOL] VRQXoODUÕQD J UH WLUDPLQL PJNJ ÕQ ]HULQGH EHOLUOHPLúOHU DUGÕQGDQ HQ \ NVHN GH HUOHU RODUDN SXWUHVLQ YH NDGDYHULQL VDSWDPÕúODUGÕU %XQD NDUúÕQ VSHUPLGLQ NRQVDQWUDV\RQXQXQ PJNJ ÕQ DOWÕQGD ROGX XQX KLVWDPLQ WULSWDPLQ YH VSHUPLQ NRQVDQWUDV\RQODUÕQÕQ LVH WHúKLV OLPLWOHULQH XODúPDGÕ ÕQÕ EHOLUOHPLúOHUGLU $\UÕFa tiramin konsantrasyonunun depolamayla önemli PLNWDUGDDUWWÕ ÕQÕELOGLUPLúOHUGLU García-García et al. \DSWÕNODUÕ odoõúpdoduõqgd WLFDUL RODUDN KD]ÕUODQDQ IDUNOÕ VRIUDOÕN]H\WLQOHULQYHGL HUWXUúXOXN JÕGDODUÕQEL\RMHQDPLQLoHULNOHULQLVDSWDPÕúODUGÕU 6RIUDOÕN]H\WLQ NHEHUHNHEHUH PH\YHVL YH KÕ\DUODUGD WRSODP EL\RMHQ DPLQ PLNWDUÕQÕQ PH\YHQLQ KHU NJ LoLQ PJ GDQ GDKD D] ROGX XQX EX QHGHQOH GH V ] JHoHQ U QOHULQ LQVDQ VD OÕ Õ LoLQ ELU ULVN ROXúWXUPDGÕ ÕQÕ LIDGH HWPLúOHUGLU <HúLO ]H\WLQ YH VDODWDOÕN UQHNOHULQLQKHSVLQGHSXWUHVLQEXOXQGX XQXDQFDNPJNJ GDQGDKDG ú N VHYL\HGHROGX XQXWHVSLWHWPLúOHUGLU < FHOYG+3/&\ QWHPLQLNXOODQDUDNJHOHQHNVHORODUDN UHWLOPLúVDODPXUDOÕYH VLWULN DVLWOH S+ VÕ D\DUODQPÕú VDODPXUDOÕ ODKDQD WXUúXODUÕQGDNL EL\RMHQ DPLQ ROXúXPODUÕQÕ LQFHOHPLúOHUGLU YH G ]H\LQGH WX] LoHUHQ WXUúXODUGDQ HQ \ NVHN EL\RMHQ DPLQ PLNWDUODUÕQÕ WX] LoHUHQ UQHNWH VDSWDPÕúODUGÕU Fermantasyonun 12. gününde % 6, 8 ve 10 düzeyinde tuz içeren sitrin DVLWOL WXUúX UQHNOHULQLQVLWULNDVLW LoHUPH\HQOHUHRUDQODGDKDD]EL\RMHQDPLQLoHUGL LQLWX] LoHUHQDVLWOLYHDVLWVL] UQHNOHUGHLVHD\QÕ LOLúNLQLQJ U OPHGL LQL WHVSLWHWPLúOHUGLU 10 düzeyinde tuz içeren örnekte fermantasyonun 12. gününde 39,8 mg/kg putresin, 8,94 mg/kg kadaverin, 88,4 mg/kg triptamin, 219,3 mg/kg spermin, 9,40 mg/kg spermidin ve 11

0,64 mg/kg tiramin belirlerken, fermantasyonun 19. gününde ise 27,4 mg/kg putresin, 71,4 mg/kg kadaverin, 29,5 mg/kg triptamin, 5,67 mg/kg spermin ve 9,85 mg/kg tiramin WHVSLW HWPLúOHUGLU 6RQXo RODUDN WX] NRQVDQWUDV\RQXQGD IHUPHQWH RODQ VLWULN DVLW LODYH HGLOPHPLú ODKDQD WXUúXODUÕQÕQ \ NVHN VHYL\HGH EL\RMHQ DPLQ ROXúXPXQD RODQDN VD ODGÕ ÕQÕEHOLUOHPLúOHUGLU âslþndet al. (2002) hücresel elektrokinetik kapiler kromatografi yöntemini kullanarak \DSWÕNODUÕ ELU odoõúpdgd KHP GR DO \ROOD KHP GH VWDUWHU NDWDUDN UHWWLNOHUL VDXHUNUDXWODUGDEL\RMHQDPLQROXúXPXQXL]OHPLúOHUGLU$OWÕDGHWNÕ\ÕOPÕúEH\D]ODKDQD ohúlglqlq ž& GD J Q V UHQ EDúODQJÕo IHUPDQWDV\RQX LOH KD]ÕUODGÕNODUÕ ODKDQD WXUúXODUÕQÕ-ž& GDGHSRODPÕúYHD\VRQXQGDDQDOL]HWPLúOHUGLU'HQHPHGHVWDUWHU kültür olarak L. plantarum (5.10 5 NREJ RODFDN úhnlogh NXOODQPÕúODUGÕU ddoõúpdgd DOÕQDQVRQXoODUDJ UHVWDUWHUNDWNÕVÕQÕQSXWUHVLQWLUDPLQYHNDGDYHULQLQ UHWLPPLNWDUÕQÕ QHPOL Oo GHD]DOWWÕ ÕQÕEHOLUOHPLúOHUGLU'L HU\DQGDQVSHUPLGLQLoHUL LQLQLOH PJNJ DUDVÕQGD GH LúWL LQL YH VSHUPLGLQ PLNWDUÕQGD VWDUWHU N OW U Q KHUKDQJL ELU D]DOPD\D QHGHQ ROPDGÕ ÕQÕ DQFDN WULSWDmin, spermin ve histamin seviyelerinin D]DOGÕ ÕQÕ YH KDWWD WHúKLV OLPLWOHULQLQ DOWÕQD LQGL LQL VDSWDPÕúODUGÕU 3UDWLNWHNL X\JXODPDODU LoLQ DUDúWÕUPDGD NXOODQÕODQ L. plantarum VXúXQXQ HQ D] 5 kob/g G ]H\LQGHVDXHUNUDXWIHUPDQWDV\RQODUÕLoLQNXOODQÕODELOHFH LVRQXFXQDYDUPÕúODUGÕU (NLFL YH &RúNXQ 9DQ LOLQGHNL PDUNHWOHUGHQ WRSODQDQ DGHW VHE]H WXUúXVX UQH LQGH DGHW NDUÕúÕN DGHW NÕUPÕ]Õ ELEHU YH DGHW KÕ\DU WXUúXVX KLVWDPLQ LoHUL LQL IOXRURPHWULN \ QWHP NXOODQDUDN LQFHOHPLúOHUGLU 7 P WXUúX UQHNOHULQGH KLVWDPLQ VHYL\HVLQLQ PJNJ LOH PJNJ RUWDODPD PJNJ DUDVÕQGD GH LúWL LQL EHOLUOHPLúOHUGLU +Õ\DU WXUúXVXQGDNL KLVWDPLQ VHYL\HVLQL PLQLPXP PJNJPDNVLPXPPJNJRUWDODPDPJNJRODUDNVDSWDPÕúODUGÕU. Moret et al. (2005) HPLC yöntemi ile taze ve konserve sebzelerde serbest biyojen amin LoHUL LQL DUDúWÕUPÕúODUGÕU 6DXHUNUDXW GÕúÕQGDNL UQHNOHUGH SXWUHVLQ -0,5 mg/100 g WD]HD ÕUOÕN YHVSHUPLGLQLQ-PJ J WD]H D ÕUOÕN HQ orn EHOLUOHQHQaminler ROGX XQXYHJHQHOOLNOHEXQODUÕVSHUPLQLQPDNVLPXPPJJWDNLSHWWL LQLWHVSLW 12

HWPLúOHUGLU.RQVHUYH VDXHUNUDXWWD WLUDPLQ VHYL\HVLQLQ PJ J ROGX XQX DQFDN GL HU UQHNOHUGHNLWLUDPLQVHYL\HVLQLQPJJ ÕJHoPHGL LQLVDSWDPÕúODUGÕU7D]H ÕVSDQDN UQHNOHULQGHLVHPJJG ]H\LQGHKLVWDPLQEHOLUOHPLúOHUGLU $NEDú +3/&\ QWHPLQL NXOODQDUDN\DSWÕ Õ odoõúpdvõqgdsl\dvdgdq WHPLQHWWL L WRSODPDGHWSDVW UL]HYHSDVW UL]HHGLOPHPLúRODUDNVDWÕODQoHúLWOLWLSWHNLWXUúXve DGHWHY\DSÕPÕWXUúX UQH LQLQEL\RMHQDPLQLoHUL LQLDUDúWÕUPÕúWÕU7XUúXODUÕQPH\YH YHYH\D VDODPXUDODUÕQGD HQ \ NVHN NRQVDQWUDV\RQD VDKLS EL\RMHQ DPLQOHUL VÕUDVÕ\OD SXWUHVLQ NDGDYHULQ YH WULSWDPLQ VÕUDVÕ\OD HY \DSÕPÕ UQHNOHUGH PJ/ 48 mg/l, 7,00 mg/kg; ticari örneklerde ise 59,69 mg/l, 18,47 mg/l, 19,51 mg/kg), en G ú NNRQVDQWUDV\RQDVDKLSEL\RMHQDPLQLLVHVSHUPLQPJ/RODUDNEHOLUOHPLúWLU 7LFDUL UQHNOHUGH HQ \ NVHN WRSODP EL\RMHQ DPLQ PLNWDUÕQÕ PJ/ HY \DSÕPÕ örnekohugh LVH PJ/ RODUDN EHOLUOHPLúWLU 6RQXo RODUDN LQFHOHQHQ W P WXUúX örneklerinde toplam biyojen amin düzeylerinin literatürde verilen ve pek çok ülkede \DVDORODUDNNDEXOJ UHQWRNVLNOLPLWOHULQDOWÕQGDEXOXQGX XQXLIDGHHWPLúWLU Günümüzde pek çrn U QGHEL\RMHQDPLQOHULQROXúXPXYHPLNWDUODUÕLOHGH LúLNWHNQLN YHIDUNOÕ\ QWHPOHULQNXOODQÕPÕNRQXVXQGD\DSÕODQoDOÕúPDODUV UPHNWHGLU g]hoolnohvrq\õoodugdjõgdodugdel\rmhqdplqohulqeholuohqphvldpdfõ\odgdkdkdvvdvyh KÕ]OÕ WHNQLNOHULQ JHOLúWLULOPHVLQH \ QHOLN odoõúpdodu GHYDP HWPHNWHGLU (HUROD et al. VÕYÕNURPRWRJUDILWHNQL LNXOODQDUDNNXUXWXOPXúVRVLVOHUGH\DSWÕNODUÕoDOÕúPDGD EL\RMHQ DPLQLQ WHúKLV OLPLWOHULQLQ WLUDPLQ VSHUPLQ VSHUPLGLQ YH NDGDYHULQ LoLQ mg/kg, feniletilamin, putresin ve triptamin için 2 mg/kg ve serotonin için de 5 mg/kg RODUDN WHVSLW HWPLúOHUGLU 6RQXo RODUDN EX \ QWHPLQ NXUXWXOPXú VRVLV UQHNOHULQGHNL EL\RMHQDPLQOHULQEHOLUOHQPHVLQGHNXOODQÕODELOHFH LQLYHD\QÕ]DPDQGDNÕ\PDSH\QLUYH 7XQDEDOÕ ÕJLELGL HUJÕGDODUÕQDQDOL]OHULQGHGHX\JXODQDELOHFH LQLEHOLUOHPLúOHUGLU Kirschbaum et al. (2000) HPLC ile biyojen aminlerin ön-kolon türevlendirmesi için 3,5- dinitrobenzoil klorür (DNBZ-&OD\ÕUDFÕQÕWHVWHWWLNOHULoDOÕúPDODUÕQGDWHúKLVOLPLWOHULQL \DNODúÕN RODrak 124- J/ HQMHNVL\RQ PLNWDUÕ - SJ EHOLUOHPLúOHUGLU 13

.DGDYHULQ YH VHURWRQLQ GÕúÕQGD EHOLUOHPH NDWVD\ÕODUÕQÕ -0,996 DUDVÕQGD EXOPXúODUGÕU %X \ QWHPL IHUPHQWH ODKDQD VX\X VR\D VRVX 0LVR VR\D H]PHVL IHUPHQWH EDOÕN VRVu ve ançüez ezmesinde agmatin, kadaverin, histamin, oktopamin, 2-feniletilamin, putresin, serotonin, spermidin, spermin, triptamin ve tiramin PLNWDUODUÕQÕQEHOLUOHQPHVLQGHX\JXODPÕúODUGÕU7 PIHUPHQWHODKDQDVX\X UQHNOHULQGH 366 mg/l putresin, 39-63 mg/l histamin, 37-73 mg/l tiramin ve 9-97 mg/l spermidin WHVSLWHWPLúOHUGLU6RQXoRODUDNEX\ QWHPLQ\ NVHNJHULG Q ú PRUDQÕYHGR UXVDOELU W UHYOHQGLUPHVD ODGÕ ÕLoLQIHUPHQWH U QOHUGHEL\RMHQDPLQG ]H\OHULQLQEHOLUOHQPHVL DPDFÕ\ODNXOODQÕODELOHFH LQLLOHULV UP úohuglu Shakila et al. EDOÕNYHEDOÕN U QOHULQGHEL\RMHQDPLQDQDOL]OHULLoLQLQFHWDEDND kromotografi (TLC) GDQVLWRPHWUL YH \ NVHN SHUIRUPDQVOÕVÕYÕ NURPRWRJUDILVL +3/& JLEL NURPRWRJUDIL WHNQLNOHULQLQ X\JXQOX X ]HULQH NDUúÕODúWÕUPDOÕ ELU odoõúpd \DSPÕúODUGÕU+3/&\ QWHPLQLQGDKDGR UXVDO-100 ng), hassas ve tekrarlanabilir (< ROPDVÕQÕQ\DQÕVÕUDNRPSOLNHDOHWOHUWHNQLNELOJLYH\ NVHNX\JXODPDPDVUDIODUÕ LOH ]DPDQ JHUHNWLUGL LQL EHOLUOHPLúOHUGLU 'L HU WDUDIWDQ GR UXVDOOÕN (20-300 ng) ve WHNUDUODQDELOLUOLNWH orn D] ELU ND\ÕSOD 7/& GDQVLWRPHWULQLQ KÕ]OÕ YH +3/& \ QWHPLQHJ UHGDKDXFX]ROGX XQXLIDGHHWPLúOHUGLU Lange et al. ohúlwoljõgd UQHNOHULQGHEL\RMHQDPLQOHULQEHOLUOHQPHVLLoLQNDSLOHU elektroforez (&(\ QWHPLLOH\ NVHNSHUIRUPDQVOÕVÕYÕNURPRWRJUDIL\ QWHPLQL+3/& NDUúÕODúWÕUPÕúODUGÕU+3/&WHNQL LLOH\ U W OHQoDOÕúPDGDWLUDPLQLoLQWHúKLVOLPLWOHULQLQ PJNJ YH &( WHNQL L LOH \ U W OHQ odoõúpdgd LVH PJNJ ROGX XQX SXWUHVLQ spermidin, kagdyhulqdjpdwlqjlelgl HUDPLQOHULQGH+3/&\ QWHPLLOH-8,5 mg/kg DUDVÕQGDVÕUDODQGÕ ÕQÕEHOLUOHPLúOHUGLU6RQXoWD&(YH+3/&DUDVÕQGDL\LELUNRUHODV\RQ ROGX XQXVDSWDPÕúODUGÕU 14

3. MATERYAL VE YÖNTEM 3.1 Materyal $UDúWÕUPDGD SL\DVDGDQ WHPLQ HGLOHQ.RUQLúRQ ohúlgl WXUúXOXN KÕ\DUODU NXOODQÕOPÕúWÕU ddoõúpdgd VWDUWHU NDWNÕOÕ UQHNOHU LoLQ NXOODQÕODQ Lactobacillus plantarum % VXúX $QNDUDhQLYHUVLWHVL0 KHQGLVOLN)DN OWHVL*ÕGD0 KHQGLVOL L% O P g UHWLPh\HVL 3URI 'U )LOL] g=d(/ø. WHQ WHPLQ HGLOPLúWLU 6DODPXUD KD]ÕUODPDGD +Õ\DU 7XUúXVX 6WDQGDUGÕ76 \H$QRQLPX\JXQWX]YHVX&D&O 2.2H 2 O (Merck), glasiyel DVHWLN DVLW 0HUFN NXOODQÕOPÕúWÕU )HUPDQWDV\RQ GHQHPHOHUL OLWUH KDFLPOL NDSDNOÕFDPNDYDQR]ODUGDJHUoHNOHúWLULOPLúWLr. 0LNUREL\RORMLN VD\ÕPODUGD HQWHUREDNWHUL YH NROLIRUP JUXEX EDNWHULOHULQ VD\ÕPÕ LoLQ VÕUDVÕ\OD 9LROHW 5HG %LOH *OXFRVH $JDU 95%* 0HUFN YH 9LROHW 5HG %LOH $JDU 95%0HUFN EHVL\HUOHULWRSODPPH]RILOLNDHURELNEDNWHUL70$%VD\ÕPÕLoLQ3ODWH Count $JDU 3&$ 0HUFN EHVL\HUL ODNWLN DVLW EDNWHULOHULQLQ VD\ÕPÕ LoLQ 0DQ 5RJRVD Sharpe Agar (MRS, Merck) besiyeri ve maya-n I VD\ÕVÕQÕ EHOLUOHPHN LoLQ 3DWDWR 'H[WURVH$JDU3'$2[RLGEHVL\HULNXOODQÕOPÕúWÕU Biyojen amin analizleri için gerekli olan tiramin (TRA), putresin (PUT), kadaverin (KAD), triptamin (TRP), feniletilamin (FEA), histamin (HIS), spermin (SPM) ve spermidin (SPD) biyojen aminleri Sigma Chemical Co. (St. Louis, MO) den VD ODQPÕúWÕU 15

3.2 Yöntem 7XUúXOXNKÕ\DUODUÕQKD]ÕUODQPDVÕ DHQHPH NXUXODFD Õ J Q SD]DUGDQ VDWÕQ DOÕQDUDN ODERUDWXYDUD JHWLULOHQ KÕ\DUODU EHNOHWLOPHGHQ D\QÕ J Q NXOODQÕOPÕúWÕU 0 PN Q ROGX XQFD HúLW E \ NO NWH VHoLOHQ KÕ\DUODU WR] WRSUDN YH \DEDQFÕ PDGGHOHU JLEL NDED SLVOLNOHULQLQ X]DNODúWÕUÕOPDVÕ DPDFÕ\ODPXVOXNVX\XLOH\ÕNDQPÕúWÕU 6WDUWHUN OW UKD]ÕUODQPDVÕ 6WDUWHU N OW U KD]ÕUODQPDVÕQGD 056 VÕYÕ EHVL\HUL YH KÕ\DU VX\X NXOODQÕOPÕúWÕU +Õ\DU VX\X HOGHHWPHN LoLQ NDEXNODUÕ VR\XOPXú KÕ\DUODU EOHQGHU\DUGÕPÕ\OD SDUoDODQPÕú YH V ]JHoWHQJHoLULOHUHNSRVDVÕD\UÕOPÕúWÕUYHRUDQODUÕQGD1D&OLODYHHGLOHQKÕ\DU VXODUÕ o & GDGDNLNDV UH\OHVWHULOL]HHGLOPLúOHUGLU Lactobacillus plantarum%vxúxghqhphqlqnxuxopdvõqgdqj Q QFHLoHULVLQGH P/056 VÕYÕEHVL\HUL EXOXQDQ W SH DúÕODQPÕú YH o C da 24 saat süreyle inkübe HGLOPLúWLU$\QÕLúOHPELUNH]GDKDWHNUDUODQPÕúWÕUøQN EDV\RQXQDUGÕQGDQ HUP/ OLN 056 VÕYÕ EHVL\HULQGH JHOLúWLULOHQ DNWLI L. plantarum 11B kültürleri içinde % 2 NaCl EXOXQDQ W SWHNL HU P/ OLN KÕ\DU VXODUÕQD DNWDUÕOPÕúWÕU7HNUDU o C da 24 saat LQN EDV\RQ VRQUDVÕQGD W S LoHUL LWRSODP P/ 1D&OLoHUHQ P/ KÕ\DU VX\XEXOXQDQP/ OLN(UOHQPD\HU HDNWDUÕOPÕúYH o C da 24 saat süreyle inkübe HGLOPLúWLU ùhnlo %X úhnlogh VWDUWHU N OW U NDGHPHOL RODUDN WX]D DOÕúWÕUÕOPÕú YH IHUPDQWDV\RQRUWDPÕQDGDKDoDEXNDGDSWHROPDVÕVD ODQPD\DoDOÕúÕOPÕúWÕU 16

ùhnlo Lactobacillus plantarum % VXúXQXQ DNWLIOHúWLULOPHVL YH KÕ\DU VX\XQD DGDSWDV\RQXQXQDúDPDODUÕ 6DIN OW U QWX]OXKÕ\DUVX\XQDDGDSWDV\RQXVÕUDVÕQGDKHUDúDPDGDVDIN OW UYH WX]OXKÕ\DUVX\X056$JDUEHVL\HULQH\D\PDN OW UHOVD\ÕP\ QWHPLQHJ UHHNLP 17

\DSÕOPÕúWÕU 3HWULOHU -30 o & GD VDDW V UH LOH LQN EDV\RQD EÕUDNÕOPÕú YH JHOLúHQ NRORQLOHUVD\ÕOPÕúWÕU% \OHFHKHPVWDUWHUN OW U QIHUPDQWDV\RQRUWDPÕQDDGDSWDV\RQX L]OHQPLúKHPGHVDODPXUD\DLODYHHGLOHQN OW UPLNWDUÕEHOLUOHQPLúWLU 6DODPXUDQÕQKD]ÕUODQPDVÕ 6DODPXUDQÕQKD]ÕUODQPDVÕQGDNXOODQÕODQPXVOXNVX\XNORUXQX]DNODúWÕUÕOPDVÕDPDFÕ\OD ND\QDWÕOÕSVR XWXOPXúWXU'HQJHDQÕQGDWX]NRQVDQWUDV\RQXQXVD ODPDNDPDFÕ\OD % 8 tuz, % 0,4 CaCl 2 YHJODVL\HODVHWLNDVLWLoHUHQOLWUHOLNLNLD\UÕVDODPXUD KD]ÕUODQPÕúWÕU6DODPXUDODUGDQELULQH GHDQODWÕOGÕ ÕúHNLOGHKD]ÕUODQDQN OW UGHQ starter kültür ordqõrodfdnúhnloghp/lodyhhgloplúwlu'l HUVDODPXUDELGRQX NRQWURO RODUDN VHoLOPLúWLU / KDFLPOL FDP NDYDQR]ODUD \DNODúÕN HU J KÕ\DU NRQXOPXú YH ]HUOHUL KD]ÕUODQDQVDODPXUDODUOD WDPDPODQPÕúWÕU KÕ\DU VDODPXUD RUDQÕ 1:1). Daha sonra turúx NDYDQR]ODUÕ RGD VÕFDNOÕ ÕQGD IHUPDQWDV\RQD EÕUDNÕOPÕúWÕU 'HQHPHNH]WHNUDUODQPÕúWÕU 3.2.4 Fermantasyonun izlenmesi ho WHNHUU UO \DSÕODQ IHUPDQWDV\RQ GHQHPHOHUL RGD VÕFDNOÕ ÕQGD ± 2 o C) \ U W OP úw U 6WDUWHU NDWNÕOÕ UQHNOHULQ KD]ÕUODQPDVÕQGa çizelge 3.1 de belirtilen günler için 11 adet kavanoz (2 adet kavanoz yedek), kontrol örneklerinin KD]ÕUODQPDVÕQGD LVH ol]hojh GH EHOLUWLOHQ J QOHU LoLQ DGHW NDYDQR] DGHW NDYDQR]\HGHNWXUúXNXUXOPXúWXU 'HQHPHGH DVHSWLN NRúXOODUÕQ NRUXQPDVÕQD ]HQ J VWHULOHUHN DOÕQDQ KÕ\DU GRNXVX YH VDODPXUD UQHNOHULQGH\DSÕODQDQDOL]OHUOHVWDUWHUOL UQHNOHULoLQYHNRQWURO UQHNOHUL LoLQ J QO N GHQHPH V UHVL LoHULVLQGH RUWD\D oõndq NLP\DVDO YH PLNUREL\RORMLN GH LúLNOLNOHU çizelge 3.1 ve çizelge 3.2 de YHULOHQ SODQODUGD J VWHULOGL L úhnlogh L]OHQPLúWLU. 18

dl]hojh6wduwhundwnõoõ UQHNOHUHDLWDQDOL]SODQÕ Analiz Titrasyon Mikrobiyolojik Biyojen amin ph Tuz günleri DVLWOL L analizler analizleri 0. + + + + +/* 1. + + +/* 2. + + + +/* 3. + + + +/* 4. + + + +/* 6. + + + +/* 8. + + + +/* 10. + + +/* 12. + + + + +/* 6DODPXUDGD\DSÕODQDQDOL]OHU +Õ\DUGRNXVXQGD\DSÕODQDQDOL]OHU dl]hojh.rqwuro UQHNOHULQHDLWDQDOL]SODQÕ Analiz Titrasyon Mikrobiyolojik Biyojen amin ph Tuz günleri DVLWOL L analizler analizleri 0. + + + + +/* 1. + + +/* 2. + + + +/* 3. + + + +/* 4. + + + +/* 6. + + + +/* 8. + + + +/* 11. + + +/* 16. + + + +/* 22. + + +/* 30. + + + + +/* 6DODPXUDGD\DSÕODQDQDOizler +Õ\DUGRNXVXQGD\DSÕODQDQDOL]OHU 3.2.5 Kimyasal analizler 6DODPXUD UQHNOHULQGH S+ WX] YH WLWUDV\RQ DVLWOL L DQDOL]OHUL TS 11112 ye (Anonim J UH\DSÕOPÕúWÕU 19

3.2.5.1 ph Salamura örneklerinde ph tayini için NEL ph 890 dijital tip phphwuhnxoodqõopõúwõu 3.2.5.2 Tuz dl]hojhyh GHYHULOHQDQDOL]SODQODUÕQDJ UHNRQWUROYHVWDUWHULODYHOL UQHNOHULQ 0, 3 ve 6. günleri ile kontrolde 30., starterli örneklerde 12. günde her bir kavanozdan DOÕQDQ P/ VDODPXUD UQH L P/ OLN(UOHQPD\HU H DNWDUÕOPÕúWÕU (UOHQPD\HU LQ LoLQHWHIORQNDSOÕNDUÕúWÕUÕFÕoXEXNDWÕOPÕúYHPDQ\HWLNNDUÕúWÕUÕFÕ ]HULQHRWXUWXOPXúWXU guqh LQ ]HULQHP/GHVWLOHVXLODYHHGLOHUHNVH\UHOWPHLúOHPL\DSÕOPÕúWÕU-2 damla metil oranj (% 0,1) indikatörü GDPODWÕOGÕNWDQ VRQUD UQH LQ Q WUDOL]DV\RQX DPDFÕ\OD VDUÕUHQNROXúDQDNDGDU-GDPOD11D2+HNOHQPLúWLU'DKDVRQUDP/ OLN K 2 CrO 4 ilave edilerek 1 N AgNO 3 LOH VDELW NLUHPLW NÕUPÕ]Õ UHQN ROXúXQFD\D NDGDU WLWUDV\RQ\DSÕOPÕúWÕU6RQXoODUWX]PLNWDUÕRODUDNKHVDSODQPÕúWÕU 7LWUDV\RQDVLWOL L 7LWUDV\RQDVLWOL LWD\LQLLoLQVWDUWHUNDWNÕOÕYHNRQWURO UQHNOHULQGHQYHJ QOHU ile starterli örneklerde 12. günde, kontrol örneklerinde 16 ve 30. günlerde örnek DOÕQPÕúWÕU $OÕQDQ P/ VDODPXUD UQH L P/ OLN (UOHQPD\HU H DNWDUÕOPÕúWÕU guqhn P/ GHVWLOH VX LOH VH\UHOWLOPLú YH PDQ\HWLN NDUÕúWÕUÕFÕ ]HULQH RWXUWXOPXúWXU Üzerine 1-2 damla fenol fitalein (% 1) indikatörü ilave edildikten sonra 0,1 N NaOH çözeltisi ile pembe renn ROXúDQD NDGDU WLWUH HGLOPLú YH VRQXoODU ODNWLN DVLW RODUDN KHVDSODQPÕúWÕU 20

3.2.6 HPLC analizi +3/&FLKD]Õ $UDúWÕUPDGD NXOODQÕODQ +3/& FLKD]ÕQÕQ ]HOOLNOHUL YH NURPDWRJUDIL NRúXOODUÕ ol]hojh WHYHULOPLúWLU dl]hojh +3/& FLKD]ÕQÕQ zellikleri ve biyojen amin analizi için kromatografi NRúXOODUÕ HPLC.RORQIÕUÕQÕ Kolon Pompa Degasser Dedektör Thermo Finnigan, Surveyor (Waltham, USA) 7KHUPR)LQQLJDQ6XUYH\RU6ÕFDNOÕNž& C-18 (250x4.6 mm, ID) Phenomenex Luna 6XUYH\RUG UWO G ú NEDVÕQoJUDGLHQWpompa Otomatik vakum degasser QP 3'$ 3KRWR 'LRGH $UUD\ SLN VDIOÕ ÕQÕQ NRQWURO LoLQ 195-QPDUDVÕQGDVSHNWUXPWDUDPDOÕEDQWJHQLúOL LQP <D]ÕOÕPSURJUDPÕ Chrom Quest Chromatography Software Mobil faz Solvent A: 30 ml buffer / 550 ml asetonitril / 420 ml destile su Solvent B: 2 ml buffer / 900 ml asetonitril / 100 ml destile su $NÕúKÕ]Õ 1,3 ml/dak. Enjeksiyon 25 µl ùhnloddoõúpdgdnxoodqõodq+3/&vlvwhpl$qndudhqlyhuvlwhvl%ø7$80 21

3.2.6.2 Biyojen amin standart çözeltilerininkd]õuodqpdvõ Tiramin (TRA), putresin (PUT), kadaverin (KAD), triptamin (TRP), feniletilamin (FEA), histamin (HIS), spermin (SPM) ve spermidin (SPD) 50 ml lik balon jojeler LoLQGH 0 SHUNORULN DVLW LOH o ] QG U OP ú YH KHU ELU EL\RMen aminin son NRQVDQWUDV\RQX PJ P/ RODFDN úhnlogh EL\RMHQ DPLQ VWDQGDUW o ]HOWLOHUL KD]ÕUODQPÕúWÕU0LNVVWDQGDUWo ]HOWLVLKD]ÕUODPDNLoLQLVHKHUELUEL\RMHQDPLQLQP/ VWRNo ]HOWLVLP/OLNEDORQMRMH\HDNWDUÕOPÕúYHKDFLP0SHUNORULNDVLW ile 50 ml \HWDPDPODQPÕúWÕU'DQVLONORU Uo ]HOWLVLLoLQPJGDQVLONORU UP/DVHWRQLoLQGH o ] QG U OP úw U7 Po ]HOWLOHUJ QO NRODUDNKD]ÕUODQPÕúWÕU% WLNRIHUet al. 1990). 3.2.6.3 Mobil faz 0WULV0DVHWLNDVLWYHGHVWLOHVXVÕUDVÕ\ODRUDQÕQGDNXOODQÕODUDNEXIIHU S+KD]ÕUODQPÕúWÕU6ROYHQW$LoLQP/EXIIHUP/DVHWRQLWULOP/GHVWLOH su ve Solvent B için de 2 ml buffer / 900 ml asetonitril / 100 ml destile su NXOODQÕOPÕúWÕU guqhnohulqkd]õuodqpdvõyhhnvwudnvl\rqx dl]hojh YH ol]hojh GH YHULOGL L JLEL IHUPDQWDV\RQXQ EHOLUOL J QOHULQGH WXUúX örneklerinin VDODPXUDODUÕQGDQ DU P/ HO UHQGHVLQGHQ JHoLULOHUHN N o N SDUoDODUD D\UÕOGÕNWDQ VRQUD HOGH HGLOHQ PH\YH NÕVÕPODUÕQGDQ LVH 10 ar grap UQHN DOÕQPÕúWÕU $OÕQDQ UQHNOHU P/ OLN (UOHQPD\HU H NRQXODUDN ]HUOHULQH µl 1-7 GLDPLQRKHSWDQ Lo VWDQGDUW 6LJPD YH P/ SHUNORULN DVLW LODYH HGLOPLú GDNLND V UH\OH HOOH odondodqpõúwõu %X úhnlogh KD]ÕUODQDQ NDUÕúÕP NDED ILOWUH ND ÕGÕQdan JHoLULOHUHNP/ OLNEDORQMRMHOHUHV ] OP úw U(OGHHGLOHQV ] QW Q Q ]HUL OLN WULNORUDVHWLNDVLW7&$LOHP/ \HWDPDPODQPÕúWÕU 22

(NVWUDNWODUÕQW UHYOHQGLULOPHVL WH DoÕNODQGÕ Õ úhnlogh KD]ÕUODQDQ UQHN HNVWUDNWODUÕQGDQ / DOÕQDUDN DO PLQ\XPIRO\RLOHVDUÕOPÕú RODQP/ OLNNDSDNOÕVDQWULI M W SOHULQHDNWDUÕOPÕú YH üzerlerine 400 µl sodyum karbonat çözeltisi ilave edildikten sonra W S NDUÕúWÕUÕFÕGD NDUÕúWÕUÕOPÕúWÕU.DUÕúÕPODUÕQ ]HULQH / GDQVLO NORU U LODYH edilip tekrar NDUÕúWÕUÕOGÕNWDQVRQUDGDNLNDV UH\OH o & OÕNVXEDQ\RVXQGDEHNOHWLOPLúWLU.DODQ GDQVLO NORU U X]DNODúWÕUPDN LoLQ / VRG\XP JOXWDPDW HNOHQPLú YH NDUÕúÕP dakika süreyle 40 o & OÕN VX EDQ\RVXQGD EHNOHWLOPLúWLU 6X EDQ\RVXQGD EHNOetme V UHFLQGHW SOHUGDNLNDDUDOÕNODW SNDUÕúWÕUÕFÕGDNDUÕúWÕUÕOPÕúWÕU%XV UHQLQELWLPLQGH 1 ml asetonitril ilave edilerek örnekler 3500 devirde 20 dakika süreyle santrifüj HGLOGLNWHQ VRQUD HOGH HGLOHQ V SHUQDWDQWODU HSHQGRUI W SOHULQH DOÕQDUDN +3/& analizi için 20 o & GDPXKDID]DHGLOPLúOHUGLU(Eerola et al. 1993). 6WDUWHUN OW U QEL\RMHQDPLQ UHWPH\HWHQH LQLQNRQWURO Lactobacillus plantarum%vxúx056võyõehvl\hulqgh o C da 24 saat süreyle iki NH] DNWLIOHúWLULOPLúWLU øoohulqgh RUDQÕQGD KLVWLGLQ WLUR]LQ WULSWRIDQ IHQLODODQLQ OLVLQ RUQLWLQ YH JOXWDPDW DPLQR DVLWOHUL EXOXQDQ P/ OLN DGHW 056 VÕYÕ EHVL\HUL LoHUHQ W SH DNWLI N OW UGHQ LQRN ODV\RQ \DSÕOPÕú YH VDDW V UH LOH o C da LQN EDV\RQD EÕUDNÕOPÕúWÕU øúohp ELU NH] GDKD WHNUDU HGLOPLúWLU VDDW VRQXQGD KHU W SWHQ HU P/ DOÕQPÕú WRSODP P/ YH P/ OLN (UOHQPD\HU HDNWDUÕOPÕúWÕU (OGHHGLOHQ NDUÕúÕPGDQVDQWULI M W S QHP/DOÕQDUDN ]HUL0 SHUNORULNDVLWOH P/ \HWDPDPODQPÕúYHGHYLUGHGDNLNDV UH\OHVDQWULI MHGLOPLúWLU(OGHHGLOHQ V SHUQDWDQWWDQYH /DOÕQDUDN WHDQODWÕOGÕ ÕJLELW UHYOHQGLUPHLúOHPL JHUoHNOHúWLULOPLúWLU7 UHYOHQGLUPHVRQXQGDHSHQGRUIW SOHUL+3/&DQDOL]LLoLQ 20 o C GDPXKDID]DHGLOPLúOHUGLU 23

3.2.7 Mikrobiyolojik analizler dl]hojh YH ol]hojh WH J VWHULOGL L JLEL IHUPDQWDV\RQXQ EHOLUOL J QOHULQGH YH VWHULONRúXOODUGDVDODPXUDODUGDQDOÕQDQP/ UQHN OLNVWHULOVHUXPIL]\RORMLNWH RUDQÕQGD VH\UHOWLOPLú YH HOGH HGLOHQ VH\UHltikler toplam mezofilik aerobik bakteriler, laktik asit bakterileri, Enterobakteri, koliform grubu bakteriler ve maya-küf VD\ÕPODUÕLoLQNXOODQÕOPÕúWÕU 7RSODPPH]RILOLNDHURELNEDNWHULVD\ÕPÕ 7RSODP PH]RILOLN DHURELN EDNWHUL VD\ÕPÕ LoLQ 3ODWH &ount Agar (PCA, Merck) EHVL\HULQH\D\PDN OW UHOVD\ÕP\ QWHPLQHJ UHHNLP\DSÕOPÕúYH-30 o C da 48 saat V UH LOH LQN EDV\RQ VRQXFXQGD JHOLúHQ NRORQLOHU VD\ÕOPÕúWÕU 6RQXoODU ORJDULWPLN GH HUOHUHoHYULOPLú YH ORJ NRE NRORQL ROXúWXUDQ ELULP P/ RODUDN LIDGH HGLOPLúWLU (Anonymous 1996). /DNWLNDVLWEDNWHULVD\ÕPÕ /DNWLN DVLW EDNWHUL VD\ÕPÕ LoLQ 056 $JDU GH 0DQ 5RJRVD 6KDUSH $JDU 0HUFN EHVL\HULNXOODQÕOPÕúWÕU<D\PDN OW UHOVD\ÕP\ QWHPLQHJ UHHNLP\DSÕODQSHWULOHU- 30 o C da 48 saat süuh LOH LQN EH HGLOPLú YH LQN EDV\RQ VRQXQGD JHOLúHQ NRORQLOHU VD\ÕOPÕúWÕU6RQXoODUORJNREP/RODUDNYHULOPLúWLU$QRQ\PRXV (QWHUREDNWHULVD\ÕPÕ (QWHUREDNWHUL VD\ÕPÕ \D\PD N OW UHO VD\ÕP \ QWHPLQH J UH 9LROHW5HG %LOH *OXFRVH Agar (VRBG,0HUFNEHVL\HULQGH\DSÕOPÕúWÕU(NLP\DSÕODQSHWULOHUGH o C da 24 saat VRQXQGD JHOLúPH ROPD\DQ SHWULOHUGH LVH VDDW VRQXQGD JHOLúHQ SHPEH-NÕUPÕ]Õ UHQNWHNL NRORQLOHU VD\ÕOPÕúWÕU 6RQXoODU ORJ NREP/ RODUDN YHULOPLúWLU $QRQ\PRXV 1996). 24

3.2.7.4 KoliIRUPJUXEXEDNWHULOHULQVD\ÕPÕ 9LROHW 5HG %LOH$JDU 95% 0HUFN EHVL\HULNROLIRUP JUXEX EDNWHULOHULQ VD\ÕPÕLoLQ NXOODQÕOPÕúWÕU<D\PDN OW UHOVD\ÕP\ QWHPLLOHHNLP\DSÕODQSHWULOHUGH o C da 24 VDDWVRQXQGD JHOLúPH ROPD\DQ SHWULOHUGH LVH VDDW VRQXQGD JHOLúHQ SHPEH-NÕUPÕ]Õ UHQNWHNL NRORQLOHU VD\ÕOPÕúWÕU 6RQXoODU ORJ NREP/ RODUDN YHULOPLúWLU $QRQ\PRXV 1996). 3.2.7.5 Maya-N IVD\ÕPÕ Toplam maya-n I VD\ÕPÕ 3RWDWR 'H[WURVH $JDU 3'$ 2[RLG EHVL\HUL NXOODQÕODUDN \D\PDN OW UHOVD\ÕP\ QWHPLQHJ UH\DSÕOPÕúWÕU%HVL\HULQLQS+ VÕQÕ HD\DUODPDN DPDFÕ\ODEHVL\HULLoHULVLQH OXNWDUWDULNDVLWo ]HOWLVLQGHQP/P/LODYH HGLOPLúWLU3HWULOHUPD\DODULoLQ-30 o C da 72 saat, küfler için ise 28-30 o C da 5 gün LQN EDV\RQD EÕUDNÕOPÕú YH GDKD VRQUD JHOLúHQ NRORQLOHU VD\ÕOPÕúWÕU 6RQXoODU ORJ NREP/RODUDNYHULOPLúWLU$QRQ\PRXV øvwdwlvwlnvhogh HUOHQGLUPH $UDúWÕUPDGD PLNUREL\RORMLN NLP\DVDO YH EL\RMHQ DPLQ DQDOL] VRQXoODUÕQD LOLúNLQ YHULOHULQ 6366 V U P SURJUDPÕ LOH DQDOL] HGLOPLú J QOHU DUDVÕQGDNL IDUNOÕOÕN LVH 'XQFDQWHVWLLOHVDSWDQPÕúWÕU' ]J QHúYG 25

%8/*8/$59(7$57,ù0$ 4.1 Starter Kültürün Salamuraya Adaptasyonu GHDQODWÕOGÕ ÕúHNLOGHKD]ÕUODQDQLactobacillus plantarum 11B starter kültürünün IHUPDQWDV\RQRUWDPÕQDDGDSWDV\RQXQXQL]OHQPHVLDPDFÕ\ODVDIN OW UGHQYHYH WX]OX KÕ\DU VXODUÕQGDQ 056 DJDU EHVL\HULQH \D\PD N OW UHO VD\ÕP \ QWHPLQH J UH HNLPL \DSÕOPÕúWÕU o & GD VDDW V UH LOH LQN EDV\RQ VRQXQGD HOGH HGLOHQ VD\ÕP sonuçlduõol]hojh GHYHULOPLúWLU Çizelge 4.1 Starter kültür adaptasyonu (log kob/ml) I. Tekerrür II. Tekerrür III. Tekerrür L. plantarum 11B 9,38 9,34 8,71 WX]OXKÕ\DU suyu WX]OXKÕ\DU suyu 9,51 9,43 9,20 9,49 8,66 8,79 L. plantarum 11B suúxqxq IHUPDQWDV\RQ RUWDPÕQD DGDSWDV\RQXQGDQ VRQUD KHU ELU WHNHUU UGHVDODPXUD\DLODYHHGLOHQVWDUWHUN OW UPLNWDUODUÕVÕUDVÕ\ODYH ORJNREP/RODUDNEXOXQPXúWXU.LP\DVDO$QDOL]6RQXoODUÕ 'HQHPHGH VWDUWHU LODYH HGLOHUHN UHWLOHQ KÕ\DU WXUúXODUÕ J Q GR DO IHUPDQWDV\RQD EÕUDNÕODUDN UHWLOHQNRQWURO UQHNOHULLVHJ QV UH\OHIHUPDQWDV\RQDEÕUDNÕOPÕúWÕU%X V UHOHULoLQGHEHOLUOLJ QOHUGHS+WLWUDV\RQDVLWOL LYHWX]PLNWDUÕQÕQEHOLUOHQPHVL DPDFÕ\ODNLP\DVDODQDOL]OHU\DSÕOPÕúWÕUdL]HOJH )HUPDQWDV\RQ HVQDVÕQGDODNWLNDVLWIHUPDQWDV\RQXQHGHQL\OH WLWUDV\RQ DVLWOL LQGHDUWÕú DVLWROXúXPXQDSDUDOHORODUDNGDS+ GDD]DOPDYHWX]PLNWDUÕQÕQGHQJH\HXODúPDVÕ\OD 26

ELUOLNWHEDúODQJÕoWDRODQWX]NRQVDQWUDV\RQXQXQ\DUÕ\DG úphvlehnohqphnwhgluøo g]oholn YH 8OX 'HQHPHGH EHQ]HU VRQXoODU HOGH HGLOPLú ROXS VRQXoODU WHNHUU U Q RUWDODPDVÕ RODUDN ol]hojh úhnlo YH úhnlo GH YHULOPLúWLU %XQD J UH EDúODQJÕo WX] NRQVDQWUDV\RQX &D&O 2 konsantrasyonu % 0,4 ve asetik asit NRQVDQWUDV\RQX RODQ VWDUWHU NDWNÕVÕ] NRQWURO UQHNOHUL LOH VWDUWHU LODYH HGLOHQ UQHNOHUH DLW VDODPXUDODUD KÕ\DUODUÕQ NRQPDVÕ\ODELUOLNWH KÕ]OÕ ELU EHVLQ YH PDGGH DOÕú YHULúLEDúODPÕúWÕU%XQDJ UHKHULNLJUXSWDGDKÕ\DUODUÕQ ]HULQGHGR DORODUDNEXOXQDQ ODNWLN DVLW EDNWHULOHUL YH NXOODQÕODQ VWDUWHU N OW U Q HWNLVL\OH WLWUDV\RQ DVLWOL L GH HUOHULQLQDUWWÕ ÕDQODúÕOPÕúWÕUdL]HOJH dl]hojh.rqwuro YH VWDUWHU N OW U NDWÕODUDN UHWLOHQ KÕ\DU WXUúXVXQGDNL NLP\DVDO GH LúLPOer 7LWUDV\RQDVLWOL L TUZ ph Gün (% Laktik asit) (%) Starter Kontrol Starter Kontrol Starter Kontrol 0. 3,51 d 3,53 d 0,41 c 0,41 c 6,06 a 5,61 a 1. 4,10 ab 4,15 ab 2. 4,18 a 4,22 a 0,41 c 0,38 c 3. 4,10 ab 4,14 ab 4,24 b 4,59 b 4. 3,72 bc 4,13 bc 0,64 b 0,54 b 6. 3,63 cd 3,80 cd 4,17 b 4,43 b 8. 3,54 d 3,73 d 0,88 a 0,75 a 10.*/11.** 3,40 d 3,61 d 12.*/16.** 3,43 3,50 0,87 0,90 4,15 22. 3,45 30. 3,45 0,98 4,20 (n=6) * Starter **Kontrol $\QÕ V WXQGD IDUNOÕ KDUIOHUOH J VWHULOHQ UDNDPODU DUDVÕQGDNL IDUN LVWDWLVWLNVHO RODUDN QHPOLGLU (P<0,01). 27

S+7X] /DNWLNDVLW * Q S+ 7X] /DNWLNDVLW ùhnlo6wduwhulodyhhglohq UQHNOHUGHIHUPDQWDV\RQV UHVLQFHS+WLWUDV\RQDVLWOL LYH WX]NRQVDQWUDV\RQXQGDJ U OHQGH LúLPOHU S+7X] /DNWLNDVLW * Q S+ 7X] 7LWUDV\RQDVLWOL L ùhnlo.rqwuro UQHNOHULQGH IHUPDQWDV\RQ V UHVLQFH S+ WLWUDV\RQ DVLWOL L YH WX] NRQVDQWUDV\RQXQGDJ U OHQGH LúLPOHU 28

6WDUWHU NDWNÕOÕ YH NRQWURO UQHNOHULQLQ VDODPXUDODUÕQGD IHUPDQWDV\RQXQ EDúODQJÕFÕQGD S+ GH HUOHUL VÕUDVÕ\OD YH RODUDN EHOLUOHQPLúWLU )HUPDQWDV\RQXQ LON J Q QGH S+ QÕQ \LQH VÕUDVÕ\OD YH \H NDGDU \ NVHOPHVLQLQ QHGHQLQLQ LVH KÕ\DUODUGDNLK FUH ]VX\XQXQVDODPXUD\DJHoPHVLROGX XG ú Q OPHNWHGLU )HUPDQWDV\RQVÕUDVÕQGDDUWDQS+GH HUOHULQLQVWDUWHUNDWNÕOÕYHVWDUWHUNDWNÕVÕ]NRQWURO UQHNOHULQGHJ QGHQLWLEDUHQD]DOGÕ ÕJ ]OHQPLúWLUdL]HOJH6WDUWHUOL UQHNOHUGH HQ\ NVHNS+ GH HULRODUDNIHUPDQWDV\RQXQ J Q QGH HQ G ú N S+ GH HULLVH 3,40 olarak fermantasyonun 10. gününde EHOLUOHQPLúWLU.RQWURO UQHNOHULQGH LVH HQ \ NVHN S+ GH HUL LOH J QGH J U O UNHQ HQ G ú N S+ GH HUL GH LOH J QGHVDSWDQPÕúWÕU S+ EDNÕPÕQGDQ \DSÕODQ YDU\DQV DQDOL]L VRQXFXQGD VWDUWHU LODYH HGLOHQ YH GR DO IHUPDQWDV\RQDEÕUDNÕODQ UQHNOHUDUDVÕQGDELUIDUNOÕOÕNJ ]OHQPHPLúWLU$QFDNJ QOHUH J UHPH\GDQDJHOHQD]DOPDQÕQLVWDWLVWLNVHORODUDN QHPOLROGX XEXOXQPXúWXU3 $UDúWÕUPDPÕ]GD VWDUWHU N OW U NXOODQÕODUDN UHWLOHQ UQHNOHUGHQ HOGHHGLOHQ S+ GH HUL KÕ\DUWXUúXVX\OD\DSÕOPÕúGL HUoDOÕúPDODUODNDUúÕODúWÕUÕOGÕ ÕQGD5RGULJRet al. (1992) XQIHUPDQWDV\RQGHQHPHVLQLQVRQJ Q RODQJ QGHEHOLUOHGL LS+\LQHEHQ]HU RODUDN \DSÕODQ )OHPLQJ et al. XQ J QO N IHUPDQWDV\RQ odoõúpdvõqõq günde ph 3,70 ve Özçelik vd. (2001) nin 10 gün süren fermantasyonun 5. günde ph 3,84-RODUDNEHOLUOHGLNOHULGH HUOHUGHQG ú NEXOXQPXúWXU)OHPLQJet al. (1973a) un 21 günlük fermantasyonun 13. gününde ph 3,38, McDonald et al. (1991) un 5 günlük fermantasyonun sonunda ph 3,42 ve Özçelik vd. (1998) nin fermantasyonun VRQ J Q RODQ J QGH S+ RODUDN EHOLUOHGL L GH HUOHUOH EHQ]HU ROGX X VDSWDQPÕúWÕU $\UÕFD )OHPLQJ et al. (1973b) un 36 gün süren fermantasyonun 13. gününde ph 3,15 ve Ulu (2001) nun fermantasyon sonu olan 12. günde ph 3,20 olarak EHOLUOHGL L GH HUOHUGHQ LVH \ NVHN ROGX X DQODúÕOPÕúWÕU 6 ] JHoHQ W P odoõúpdodugd WDPSRQNXOODQÕOPÕúWÕU 29