Characteristics of the Leuconostoc mesenteroides subsp. mesenteroides strains from fresh vegetables

(1)

UDC635.1/.6:579.64:579.8 APTEFF, 37,1-192(2006) BIBLID: 1450–7188 (2006) 37,3-11 Original scientiic paper

3 'U*RUGDQD5'LPLü$VVRF3URI8QLYHUVLW\RI1RYL6DG)DFXOW\RI7HFKQRORJ\1RYL6DG%XOHYDU &DUD/D]DUD6HUELD

CHARACTERISTICS OF THE

Leuconostoc mesenteroides

subsp.

mesenteroides

_{STRAINS FROM FRESH VEGETABLES}

*RUGDQD5'LPLü

Strains synthesizing extracellular polysaccharide dextran on a medium with 10%

VXFURVHZHUHLVRODWHGIURPGLIIHUHQWNLQGRIYHJHWDEOHVFDEEDJHFXFXPEHUFDXOLÀRZHU

kohlrabi, carrot, green beans, red beet, pepper, eggplant, radish). Carbohydratefermentationbohydratefermentationfermentation was examined using a bioMerieux API 50 CHL test system. Among micropopulations with characteristic spherical cell morphology, 94.9% belonged to Leuconostoc mesenteroides subsp. mesenteroides and 5.1% were identifíed as Leuconostoc mesenteroides subsp. dextranicum. According to fermentation of pentoses L. mesenteroides strains were divi-ded into three groups with a certain number of biotypes; 10 strains were tested on acid production.

.(<:25'6LeuconostocYHJHWDEOHODFWLFDFLGEDFWHULD FDUERK\GUDWHPHWDEROLVPIHUPHQWDWLRQ

,1752'8&7,21

7KHIHUPHQWDWLRQRIYHJHWDEOHVLVLQRXUFRXQWU\VWLOOSHUIRUPHGLQWKHWUDGLWLRQDOZD\ VSRQWDQHRXVIHUPHQWDWLRQKRZHYHUWKHTXDOLW\RIWKH¿QLVKHGSURGXFWLVQRWXQLIRUP 9HU\RIWHQGXHWRORZLQLWLDOQXPEHURIODFWLFDFLGEDFWHULDRUWKHLUORZDFWLYLW\DQGLP

-SRVVLEOHFRQWUROWKHUHVXOWRIWKHSURFHVVLVXQFHUWDLQ

6HYHUDONLQGVRIIDFXOWDWLYHO\DQDHURELFODFWLFDFLGEDFWHULDSDUWLFLSDWHLQWKHIHUPHQ

-WDWLRQRIYHJHWDEOHV$PRQJWKHVHEDFWHULDLeuconostoc mesenteroidesVXEVSmesenteroi-desLactobacillus plantarum and_L. _brevisDUHWKHPRVWLPSRUWDQW7KH\DUHQDWXUDOO\

SUHVHQWRQYHJHWDEOHV

(2)

4

SRO\PHUL]DWLRQRIJOXFRVHXQLWVIURPVXFURVH0XFKPRUHLPSRUWDQWKDVWKLVJURXSRI PLFURRUJDQLVPVLQODFWLFDFLGIHUPHQWDWLRQRIIRRGWRJHWKHUZLWKRWKHUODFWLFDFLGEDFWHULD

+HWHURIHUPHQWDWLYHLeuconostoc mesenteroidesVXEVSmesenteroidesIXUWKHUL. me-senteroidesGLVLQWHJUDWHVJOXFRVHWR'ODFWLFDFLGDQGLVWKHPRVWUHVSRQVLEOHIRUWKH FUHDWLRQRIDQDHURELFFRQGLWLRQVHVSHFLDOO\LQWKHLQLWLDOSKDVHRIIHUPHQWDWLRQDQGLWV SURSHUGHYHORSPHQW

7KHDLPRIWKHZRUNZDVWRGHWHUPLQHWKHVWUDLQVRIL. mesenteroidesDQGWROHDUQ PRUHDERXWWKHLUSKHQRW\SHFKDUDFWHULVWLFVFRQFHUQLQJWKHUROHRIWKHVHEDFWHULDLQSURGXF

-WLRQRIIHUPHQWHGYHJHWDEOHV

(;3(5,0(17$/

Isolation procedure

7KHVDPSOHVRIIUHVKYHJHWDEOHVJUHHQEHDQVFDXOLÀRZHUNRKOUDELFDEEDJHFXFXP

-EHUFDUURWHJJSODQWSHSSHUUDGLVKDQGUHGEHHWZHUHPDVKHGDQGVXVSHQGHGLQWROLTXLG PHGLXPZLWKVXFURVHPODQGLQFXEDWHGIRUWRKRXUVDWWKHWHPSHUDWXUHRI &,QWKHSUHVHQFHRIGH[WUDQRJHQRXVEDFWHULDWKHPHGLXPEHFDPHWKLFNDQGODFWLF7KH VHOHFWLYHPHGLXPFRQWDLQHGJOFRQVXPHVXFURVHJ\HDVWH[WUDFWJ.+324J

1+4624J0J624× 7H2J1D&OJ3XUHFXOWXUHVZHUHREWDLQHGRQDJDU

PHGLXPRIWKHVDPHFRPSRVLWLRQ

&ODVVL¿FDWLRQDQGFKDUDFWHUL]DWLRQRILVRODWHV

7KH VWUDLQV ZHUH GHWHUPLQDWHG E\ WKH SURFHGXUH GHVFULEHG E\ *DUYLH 6HYHUDO SKH7KHVWUDLQVZHUHGHWHUPLQDWHG E\ WKH SURFHGXUH GHVFULEHG E\ *DUYLH 6HYHUDO SKHGHWHUPLQDWHGE\WKHSURFHGXUHGHVFULEHGE\*DUYLH6HYHUDOSKH

-QRW\SHFKDUDFWHULVWLFVLQFOXGLQJFHOOPRUSKRORJ\FDWDODVHDFWLYLW\&2SURGXFWLRQIURP JOXFRVHDQGGH[WUDQSURGXFWLRQZHUHVWXGLHG7KHJURZWKLQWKHSUHVHQFHRI1D&O ZHUHWHVWHGLQGHHSDJDU

3XUHFXOWXUHZHUHFXOWLYDWHGRQ056DJDUIURPODFWLFDFLGEDFWHULDLQRUGHUWRLQYHV

-WLJDWHFDUERK\GUDWHVIHUPHQWDWLRQ)RUWKLVSXUSRVHWKH$3,&+/WHVWZDVXVHGJLYLQJ WKHSRVVLELOLW\WRWHVWGLIIHUHQWVXEVWUDWHV,QRFXOXPZDVSUHSDUHGZLWKDFXOWXUHGLOXWHG WRPDWFKD0F)DUODQGWXUELGLW\VWDQGDUG$3,6\VWHP6$/D%DOPHOHV*URWWHV 0RQWDOLHX9HUFLHX)UDQFH$OOWHVWVZHUHUHDGDIWHUDQGKRXUVRILQFXEDWLRQDW &

Acid production.

7KHFKRVHQVWUDLQVRIL. mesenteroidesZHUHSODWHGLQOLTXLGPHGLXPIRUWKHIROORZLQJ RIDFLGSURGXFWLRQZKLFKFRQWDLQHGJOWULSWRQJ\HDVWH[WUDFWJJOXFRVHJ VDOWVROXWLRQ$DQG%POVDOW$-K

+324.+324VDOW%-0J624× 7HO

1D&O)H6240Q624× 4H2'XULQJVHYHQGD\VRILQFXED

(3)

5(68/76$1' ',6&866,21$1'',6&866,21

)URPGLIIHUHQWNLQGVRIYHJHWDEOHVLeuconostocVWUDLQVV\QWKHVL]LQJPXFRXVGH[

-WUDQRQVXFURVHPHGLXPZHUHLVRODWHG7KHSURGXFWLRQRIGH[WUDQVLVVLJQL¿FDQWRQO\IRU WZRVSHFLHVRIWKLVJHQXVL. mesenteroidesand_{L. dextranicum}2QVXFURVH

DJDUWKHLVRODWHVIRUPHGVHYHUDOW\SHVRIPXFRXVFRORQLHVEXWLQJHQHUDOWZRW\SHVFRXOG EHGH¿QHGDFRORQLHVZLWKFOHDUO\H[SUHVVHGPDFURPRUSKRORJLFDOFKDUDFWHULVWLFVFRQ

-YH[ZLWKÀDWHGJHVVPRRWKVKLQ\VHPLWUDQVSDUHQW)LJDDQGEVSUHDGFRORQLHV)LJ E,WLVNQRZQWKDWGH[WUDQVREWDLQHGZLWKGLIIHUHQWVWUDLQVRIWKHVDPHVSHFLHVDUHQRW LGHQWLFDOLQFKHPLFDOVWUXFWXUH

Fig. 1. L. mesenteroidesRQVXFURVHDJDU

DFRPSDFWFRORQLHVEVSUHDGFRORQLHV

7KHGLIIHUHQFHLQVL]HRIFRORQLHVZDVSDUWLFXODUO\H[SUHVVHGIRULQGLYLGXDOVWUDLQV 'LDPHWHURIFRORQLHVUDQJHGIRUWKUHHGD\VFXOWXUHVIURPWRPPLQDYHUDJH6SKHUL

-FDOO\VKDSHGFHOOVDUHJURXSHGPRVWO\LQSDLUVVKRUWHUDQGORQJHUFKDLQV)LJDZKHUHDV DWFHUWDLQVWUDLQVWKHDJJUHJDWLRQRIFHOOV)LJELVQRWLFHDEOH

$OOVWUDLQVVKRZHGQHJDWLYHFDWDODVHWHVWDQGSURGXFHGJDVHVIURPJOXFRVHDQG JURZLQWKHSUHVHQFHRI1D&O&RQVLGHULQJWKHDELOLW\WRGLVLQWHJUDWHSHQWRVHWKHVH EDFWHULDSRVVHVVHGWKHFKDUDFWHULVWLFVRIL. mesenteroidesDVDNH\IRUFODVVL¿FDWLRQDQG GLIIHUHQWLDWLRQRIPXFXVSURGXFLQJLeuconostocVDUDELQRVHZDVXVHG2QO\ RIVWUDLQVLVRODWHGIURPYHJHWDEOHVZHUHQRWDEOHWRXVHDUDELQRVHDQGGLGQRWJURZLQWKH SUHVHQFHRIKLJKHUVDOWFRQFHQWUDWLRQV7KH\EHORQJHGWRWKHOHVVWROHUDQWL. dextranicum.

,QRUGHUWRLPSURYHSKHQRW\SLFVSHFLHVFKDUDFWHUL]DWLRQRIWKHJHQXVLeuconostoc 0LOOLHUHHWDOUHSRUWHGWKDWWKHWROHUDQFHWR1D&OYDULHGH[FHSWIRUL. cremoris ZKLFKFRXOGQRWJURZDWWKLVFRQFHQWUDWLRQ/DFWLFVWUHSWRFRFFLZKLFKDUHWD[RQRPLFDOO\ DQGHFRORJLFDOO\FRQQHFWHGZLWKOHXRFRQRVWRFVDUHH[FOXGHGEHFDXVHWKH\GRQRWGLVLQ

(4)

Fig. 2. L. mesenteroides PLFURPRUSKRORJLFDOFKDUDFWHULVWLFî

DFKDLQVEDJJUHJDWLRQ

7KHLQYHVWLJDWLRQVRIFDUERK\GUDWHPHWDEROLVPVXVLQJELR0HULHX[$3,V\VWHPEDVHG RQIHUPHQWDWLRQRISHQWRVHVDUDELQRVH[\ORVHDQGULERVHUHVXOWHGLQGLYLGLQJWKHVWUDLQV RIL. mesenteroidesLQWRJURXSVZLWKFHUWDLQQXPEHURIELRW\SHV7KHUHVXOWVDUHSUH

-VHQWHGLQ7DEOHVDQG

*URXS$FRQVLVWVRIELRW\SHVZLWKWRWDOO\VWUDLQV7DEOH7KHFKDUDFWHULVWLF RIWKLVJURXSLVWKDWDOOVWUDLQVVKRZHGSRVLWLYHUHDFWLRQWRIHUPHQWDWLRQRI/DUDELQRVH &RQVLGHULQJWKHQXPEHURIVXEVWUDWHVWKH\GLVLQWHJUDWHIROORZLQJELRW\SHV$DUHGLVWLQ

-JXLVKHGJDODFWRVHPDQQLWRODP\JGDOLQDQG$PDQQLWRODP\JGDOLQODFWRVH$OOWKHVH UHDFWLRQVZHUHQHJDWLYHIRULVRODWHV$

Table 1.'LIIHUHQWLDWLQJFKDUDFWHULVWLFVRIL. mesenteroidesVWUDLQVJURXS$

Characteristic A1 A2 A3 A4 A5

*DODFWRVH + + – – –

0DQQLWRO + – – + +

$P\JGDOLQ + + – + +

/DFWRVH – – – – +

'5D¿QRVH – + – – –

*URXS%LVFKDUDFWHUL]HGE\DVPDOOHUQXPEHURIVWUDLQVEXWE\DKLJKHUYDULDELO

(5)

7

Table 2. 'LIIHUHQWLDWLQJFKDUDFWHULVWLFVRIL. mesenteroidesVWUDLQVJURXS%

Characteristic B1 B2 B3 B4 B5 B6

0DQQLWRO – – + – – –

$P\JGDOLQ + + – – – –

$UEXWLQ + + – – + –

6DOLFLQ + + + – + +

&HOORELRVH + + + – – –

/DFWRVH + – + + – –

ȕ*HQWLELRVH + + + + –

*OXFRQDWH – + + – – –

NHWR

JOXFRQDWH – – + – – –

7KHODVWJURXS&LQFOXGHGWKHVWUDLQVIHUPHQWLQJ/DUDELQRVHDQG'[\ORVH)LYH VXFKVWUDLQVZHUHLGHQWL¿HG7DEOH$FFRUGLQJWRFKDUDFWHULVWLFVWKH\ZHUHJURXSHGLQ WKUHHELRW\SHVDQGWKHPRVWDFWLYHRQHZDV&

Table 3. 'LIIHUHQWLDWLQJFKDUDFWHULVWLFVRIL. mesenteroidesVWUDLQVJURXS&

Characteristic C1 C2 C3

$P\JGDOLQ – + –

$UEXWLQ + + –

6DOLFLQ – + –

&HOORELRVH + + –

0HORELRVH – + +

'5DI¿QRVH – + +

*OXFRQDWH + – –

$P\JGDOLQVKRZHGWREHPRVWO\IHUPHQWHGE\DOOWKUHHJURXSVL. mesenteroides7D

(6)

8

Table 4. 1XPEHURISRVLWLYHVWUDLQVZLWKIHUPHQWDWLRQDELOLW\RILVRODWHGL. mesenteroidesJURXSV

Source of fermentation

Group

A B C

$P\JGDOLQ 3

$UEXWLQ 4

&HOORELRVH 4

'5DI¿QRVH 4

*DODFWRVH

*OXFRQDWH 8

/DFWRVH

0DQQLWRO

0HORELRVH 4

6DOLFLQ 3

E*HQWLELRVH

NHWRJOXFRQDWH

Table 5. )UHTXHQFHRIL. mesenteroidesJURXSVLQYHJHWDEOHV

Vegetable Group [%]

A B C

3HSSHU

&DXOLÀRZHU

&DEEDJH

*UHHQ%HDQ

&XFXPEHU

(JJSODQW

5DGLVK

5HG%HHW

.RKOUDEL

&DUURW

(7)

Table 6. $FLGSURGXFWLRQDQGS+YDOXHVRIL. mesenteroides

DWWDLQHGDIWHUJURZWKIRUDQGGD\V

Strain 1. day 7. day

Acid (%) pH Acid (%) pH

$

%

,QLWLDOS+

5HJDUGLQJWKHIHUPHQWDWLYHDFWLYLW\¿YHVWUDLQVDUHVWDQGLQJRXWZLWKKLJKHVWSURGXF

-WLRQRIDFLG$%DQG$$$6WUDLQ$SUR

-GXFHGWKHORZHVWDPRXQWRIDFLGGXULQJWKHVHYHQGD\VRIIHUPHQWDWLRQ±

&21&/86,21

5HVHDUFKRIIHUPHQWDWLRQIHDWXUHVRIODFWLFDFLGEDFWHULDLVWKH¿UVWVWHSLQWKHVHOHFWLRQ RIVWUDLQVFDSDEOHWRSURGXFHVXI¿FLHQWDPRXQWRIDFLG7KHDLPRIIXUWKHULQYHVWLJDWLRQ ZDVWRFKRRVHWKHPRVWVXLWDEOHVWUDLQVRIL. mesenteroides DVVWDUWHUFXOWXUHVIRUWKHFRQ

-WUROOHGIHUPHQWDWLRQRIYHJHWDEOHV

5()(5(1&(6

)DXULH-0-=/HYHDQDQG0%RXL[,QÀXHQFHRIGH[WUDQVRQWKHDGKHVLRQRI Leuconostoc mesenteroidesKH[DGHFDQH6FLHQFHVGHV$OLPHQWV17 7DOOJUHQ$+8$LUDNVLQHQ5YRQ:HLVVHQEHUJ+2MDPR-.XXVLVWRDQG0

/HLVROD([SRO\VDFFKDULGHSURGXFLQJEDFWHULDIURPVXJDUEHHWV$SSOLHGDQG(QYL

-URQPHQWDO0LFURELRORJ\65

&KHQ.+5)0F)HHWHUVDQG+3)OHPLQJ)HUPHQWDWtRQFKDUDFWHULVWLFVRI KHWHURODFWLFDFLGEDFWHULDLQJUHHQEHDQMXLFH-RXUQDORI)RRG6FLHQFH48

$QGHUVVRQ5&KDUDFWHULVWLFVRIWKHEDFWHULDOÀRUDLVRODWHGGXULQJVSRQWDQHRXVODF

(8)

1RZDNRZVND%DQG-/LSRYVNL0DQXIDFWXUHRIIHUPHQWHGYHJHWDEOHEHYHUDJHVXV

-LQJGLUHFWHGODFWLFIHUPHQWDWLRQ3U]HP\VOIHUPHQWDF\MQ\L2ZRFRZR:DU]\ZQ\39

6RRQ'RQJ..0HH.\XQJDQG.6RR.X(IIHFWRIGLIIHUHQWVDOWFRQFHQWUDWLRQV DQGWHPSHUDWXUHVRQWKHODFWLFDFLGIHUPHQWDWLRQRIUDGLVKMXLFH-RXUQDORI)RRG6FL

-HQFHDQG1XWULWLRQ4

0RUHD0)%DP]]LDQG36&RFFRQFHOOL0ROHFXODUDQGSK\VLRORJLFDOFKDUDFWHU

-L]DWLRQRIGRPLQDQWEDFWHULDOSRSXODWLRQVLQWUDGLFLRQDO0R]]DUHOODFKHVVHSURFHVV

-LQJ-RXUQDORI$SSOLHG0LFURELRORJ\87

/RSH]'LD]70&5$ORQVR0/*DUFLD/RSH]DQG%0RUHQR/DFWLFDFLG EDFWHULDLVRODWHGIURPKDQGPDGHEOXHFKHHVH)RRG0LFURELRORJ\17 *DUYLH(,*UDPSRVLWLYHFRFFL*HQXVLeuconostocLQ%HUJH\V0DQXDORI6LVWH

-PDWLF%DFWHULRORJ\9RO(GV3+$6QHDWK160DLUDQG0(6KDUSH:L OOLDPV:LONLQV%DOWLPRUHS

'RPDQ . DQG - ( 5RE\W 'H[WUDQVXFUDVH FRQVWLWXWLYH PXWDQWV RILeuconostoc mesenteroides%(P]\PHDQG0LFURELDO7HFKQRORJ\17 6KDPDOD75DQG063UDVDG3UHOLPLQDU\VWXGLHVRQWKHSURGXFWLRQRIKLJKDQG

ORZ YLVFRVLW\ GH[WUDQ E\Leuconostoc VSS 3URFHVV %LRFKHPLVWU\30

<XVHII++6$(O$VVDUDQG60))DWK\2SWLPL]DWLRQRIFXOWXUHFRQGLWLRQV IRU WKH SURGXFWLRQ RI GH[WUDQ E\Leuconostoc mesenteroides $GYDQFHV LQ )RRG 6FLHQFHV19

6KLQ&DQG<6DWR,GHQWL¿FDWLRQRILeuconostocVWUDLQV-SQ-RXUQDORI=RRWHFK

6FLHQFH52

0LOOLHUH-%$*0DWKRW36FKPRWWDQG&'LYLHV3KHQRW\SLFFKDUDFWHUL]DWLRQ RILauconostocVSHFLHV-RXUQDORI$SSOLHG%DFWHULRORJ\67 9DOGH]*)*6GH*LRUL0*DUUR)0R]]LDQG*2OLYHU/DFWLFDFLGEDFWHULD

IURP QDWXUDOO\ IHUPHQWHG YHJHWDEOHV 0LFURELRORJLH $OLPHQWV 1XWULWLRQ8

ɄȺɊȺɄɌȿɊɂɋɌɂɄȿɋɈȳȿȼȺ

Leuconostoc mesenteroides

subsp

. mesenteroides

ɂɁɈɅɈȼȺɇɂɏɂɁɋȼȿɀȿȽɉɈȼɊȶȺ

ȽɨɪɞɚɧɚɊȾɢɦɢʄ

(9)

ɦɚɮɟɪɦɟɧɬɚɰɢʁɢɩɟɧɬɨɡɚL.mesenteroidesɫɨʁɟɜɢɫɭɫɜɪɫɬɚɧɢɭɬɪɢɝɪɭɩɟɫɚɨɞɝɨ

ɜɚɪɚʁɭʄɢɦɛɪɨʁɟɦɛɢɨɬɢɩɨɜɚɫɨʁɟɜɚL. mesenteroidesʁɟɬɟɫɬɢɪɚɧɨɧɚɩɪɨɢɡɜɨɞʃɭ ɤɢɫɟɥɢɧɟ