PHN I.

GII THIU
Ngun gc ca qu trnh bo qun rau qu bng phng php ln men c t thi c i. Trn th gii, hu ht rau qu c ln men vi qui m nh, nh trong iu kin gia nh hoc bi nhng ngi bun bn nh. Hin nay c mt s mt hng sn xut dng ny nh da bp ci, da chut mui chua v liu mui chua. C ngha thng mi quan trng v l tiu im cho seminar ny. C s gia tng quan tm trong phm vi rng cc loi rau qu ln men khc v nc p rau qu ln men , c bit th trng Chu u (Bng 2.1); tuy nhin, cc dng thng mi hu ht c sn xut vi qui m nh (Buchenhuskes cng cng s, 1990). Kimchi Hn Quc, mang tnh truyn thng trong cc ba n gia nh, gn y cng c thng mi ho (Cheigh & Park, 1994; Fleming cng cng s, 1995a). Vic nghin cu vi sinh vt trong qu trnh ln men rau qu sm bt u vo nhng nm u 1900. Nhiu nghin cu vn c gi tr n ngy nay, nhng c nhiu thay i trong qu trnh ln men rau qu vi qui m thng mi. Tiu im nghin cu gn y l vic rt b nc mui, l vn c ngha mi trng, v p dng vic cy men ging iu khin qu trnh ln men sn phm cui cng tng tnh vng chc. Nhiu bo ch vit v ch ln men rau qu, v ngi c c ch dn thm thng tin v ch ny. (Daeschel cng cng s, 1987; Buchenhuskes cng cng s, 1990; Fleming cng cng s, 1995a; Garrido-Fernandez cng cng s, 1995). Vi sinh vt tiu biu ca rau qu ti b nh hng ln bi vi khun gram m hiu kh v nm men, trong khi mt vi khun lactic to thnh lc u l th yu (Mundt cng cng s, 1967; Mundt & Hammer, 1968; Mundt, 1970; Schneider, 1988). Tuy nhin, hu ht rau qu hoc nc p ca chng chu s ln men lactic t pht trong iu kin ym kh vi m, nng mui v nhit c iu chnh thch hp vi khun lactic c s cnh tranh thun li. S pht trin ca vi khun lactic (Bng 2.2) ph thuc vo thnh phn ho hc (mi trng dinh dng, nng mui, pH) v tnh cht vt l (loi rau qu, nhit mi trng). Mi trng thay i trong sut qu trnh ln men, sinh vt chim u th thng l vi khun c c im c bit v sinh trng tt. Vi khun gram m sm b c ch trong qu trnh ln men bi mui thm vo v acid to thnh nhanh. Bng 2.1 Mt vi sn phm rau qu ln men mang tnh thng mi Chu u, Bc M v Hn Quc. Sn phm thng mi chnh Sn phm thng mi ph liu Artis C chua xanh Da bp ci Bp ci (C chua cha chn) Da leo Capers Kimchia C rt Lupinus beans Ci bng Da Cn ty t ngt C tm t cay Ti C ci ng u xanh Bp ci C ci Trch t Buchenhiskes, 1993. a Sn phm c tnh thng mi cao Hn Quc.
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Bng 2.2 Sn phm phc tp ca vi khun lactic trong qu trnh ln men rau qu.
Vi khun acid lactic Enterococcus faecalis Lactobacillus bavaricus Lactobacillus brevis Lactobacillus plantarum Lactococcus lactis Leuconostoc mensenteroides Pediococcus pentosaceus Dng ln men Ln men ng th Ln men ng th Ln men d th Ln men ng th Ln men d th Ln men ng th Ln men d th Ln men ng th Cc sn phm chnh Lactate Lactate Lactate, acetate, CO2 Lactate Lactate, acetate Lactate Lactate, acetate, CO2 Lactate Hnh dng lactic L(+) L(+) DL D(-),L(+), DL D(-),L(+), DL L(+) D(-) DL, L(+)

Trch t Fleming cng cng s (1995a). Vi khun lactic c kh nng sinh ra cc cht c hot tnh khng sinh, c ch vi khun phn hu peptides hoc protein tu thuc vo ging loi. Vi khun lactic l loi ph bin sn sinh ra cc cht c hot tnh khng sinh c trong qui m ln v nh (Klaenhammer, 1988). Nhng kt qu bo co nghin cu trn qui m ln, sn phm cht c hot tnh khng sinh ca vi khun lactic c phn lp t qu trnh ln men rau qu t nhin biu th rng cht khng vi khun phn hu protein ng vai tr quan trng trong qu trnh ln men thc phm truyn thng v mt sinh thi hc. (Fleming cng cng s, 1975; Daeschel & Klaenhammer, 1985; Anderson, 1986; Daeschel cng cng s, 1990; Harris cng cng s, 1992a; Uhlman cng cng s, 1992; Atrih cng cng s, 1993; Jimenez-Diaz cng cng s, 1993).

PHN II. CC DNG SN PHM LN MEN T VI SINH VT.

2.1. DA BP CI S tin ho ca sn phm bp ci mui chua c bit n ngy nay l do nghin cu ca Pederson (1960) v Pederson & Albury (1969). Bp ci (Brassica oleracea) c bit v s dng thng t nht 4000 nm. Da bp ci l kt qu ca sn phm t qu trnh ln men lactic t nhin ca mui, bp ci c thi si. Theo ngi c ngha ca t acid (lm chua / Sauer) bp ci (Braut). u tin da bp ci c lm t nhng l bp ci vi ngun gim chua hoc gim cng nghip (Vaughn, 1985). Sau , bp ci c ct thnh nhiu ming nh, cho vo thng v rt dung dch c tnh acid Nc qu chua t cn xanh (cha chn) nho hoc ngun tri cy khc. Thi gian chuyn i t dung dch c tnh acid thnh nc da mui th khng c nh. 2.1.1.Nguyn liu Loi bp ci (cy trng) sn xut da bp ci thng mi c pht trin mt cch c bit. Yu cu l bp ci c mu trng, hng v du, ngt, v n lm cho sn phm tt hn. im cn quan tm l rng ln v dy c phn u (3.5- 5.5Kg) vi s lng ti thiu l xanh pha ngoi. Nng ng c th ln men c khong 5%, vi nng fructose v glucose xp x ngang bng nhau (mi loi 2.5%) v lng ng sucrose thp (Bng 2.3). Thnh phn ca mt vi loi bp ci c cht dit khun v n ph thuc vo loi vi khun lactic chim u th trong qu trnh ln men (Kyung & Fleming, 1994a). Hu ht nhng thnh phn km hm quan trng l sn phm thy phn ng glucose to
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thnh iso thiocyanates, nitriles v thicyanates.Gn y, hp cht km hm methyl methanethiosulphinates c xem nh hp cht dit khun ch yu trong bp ci (Kyung & Fleming, 1994b). Bng 2.3 Hm lng ng kh v khng kh trong nguyn liu bp ci, da chut v liu. Hm lng (%) Thnh phn a L bp ci Li bp cia Da chutb liuc ng khng kh Sucrose 0.1- 0.4 1.3- 2.4 NDd 0.1- 0.2 ng kh 2.0- 2.7 Fructose 2.0- 2.1 1.0- 1.2 1.0- 1.4 0.1- 1.1 Glucose 2.2- 2.6 1.1- 1.7 1.0- 1.3 1.0- 3.0
a

Trch t Fleming cng cng s (1988b). Trch t McCombs cng cng s (1976) v Fleming cng cng s (1988b). c Trch t Garrido- Fernandez cng cng s (1995). d ND, khng pht hin (khng tm thy).
b

2.1.2.Cng ngh ch bin Da bp ci c lm t phng php mui kh bp ci (Hnh 2.1). Bp ci t c ct loi b phn l xanh bn ngoi, l gy hoc l d (ti 30% khi lng). Nhng phn li c ct bng my ct li o ngc tc l ct phn li trn u ca l. iu chnh dao quay ct u bp ci thnh nhng ming nh c kch thc 0.08- 0.16 cm. Tt hn l ct thnh si di nhng dy c xc nh tu thuc vo ngi sn xut, my ct nh v bp ci c s dng. Bp ci ct (cng nh mn x lch t bp ci ) c chuyn bng bng ti n nhng thng cha.Tin trnh mui khc nhau. nhng ni c phng tin sn xut th bp ci ming nh c cn trn dy bng ti v ng lng mui (2- 2.5%) c rc trn bp ci ming nh khi khi bp ci di chuyn dc bng chuyn n thng cha. nhng ni khc, vn chuyn bng tay th bp ci c th c cn mi ln vn chuyn hoc cn nh k bp ci trong thng cha kim tra s lng s lng chuyn n thng cha.Trong qu trnh ch bin, cn quan tm lng mui vo c ng u. Nhng ci co phn phi nhng ming ci nh cht thnh ng trong thng cha ngm mui ng u. Vi nhng thng ln men ln 180 tn, nhng tm thu tinh c s dng ngn cch mi ngn khong 20 tn; tuy nhin, nhiu thng ln men nh c s dng trong sn xut da bp ci c tnh c bit. Mi thng cha c y n mc thch hp, ph mt tm plastic ln che trn nh thng cha. Trong giai on u, mui lm cho dch bo trong rau tit ra. Trng lng dung dch nc v nc mui pha trn nn p ming bp ci nh i xung n khi nhng ming bp ci c ngp ht trong nc mui. Trong thi gian ngn, iu kin trong thng cha tr nn ym kh bi oxy gim v CO2 sinh ra do s pht trin ca vi khun lactic. Nhit v nng mui NaCl l tng cho qu trnh ln men da bp ci 0 khong 18 C v 1.8- 2.25% NaCl. iu ny c Parmele cng cng s (1927) ghi nhn u tin v Marten et al (1929), v c trnh by chi tit trong nghin cu ca Pederson & Albury (1969). Khi gn nhit v nng mui ny th vi khun lactic bt u hot ng (Hnh 2.2). Thc t sn xut thng mi Bc M khng iu khin nhit , qu trnh ln men xy ra nhit mi trng xung quanh. Nhit ca qu trnh ln men
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trong lc u lin quan n nhit bp ci nhp vo, c th thay i (t thp hn 10 0C n xp x 300C), ph thuc vo thi gian v thi tit trong nm. Phng php p mui kh lm cho nng mui khng ng u trong thng cha.(Pederson & Albury, 1969). Qu trnh ln men c tin hnh ngn l khong mt vi tun, di khong mt nm trc khi bao gi. Bc M, da bp ci c d tr trong thng ln men v ng hp khi n t hng ca khch hng. y l ng li kinh t tn tr da bp ci nhng kt qu l lm thay i sn phm cui. Nng acid lactic cao trong sn phm da bp ci trong qu trnh bo qun c th lm cho nng mui thay i hoc long, lm gim mi v v gy ra s h hng. Chu u, bao b v thanh trng sn phm c bn da trn pH, tnh acid, hoc nh gi cm quan, ta sn phm ng nht hn, hng v hi ho hn.

Da bp ci c th c bao gi trong nhng ci hp, bnh thu tinh hoc bao plastic (Fleming cng cng s, 1995a). Da bp ci c lm lnh ( v bao b thu tinh hoc plastic) l sn phm khng c thanh trng, c thm cht bo qun nh Natri benzoate (0.1% w/w) v Kali metabisulphite. C th lm n nh da bp ci bng phng php thanh trng nhit 74 - 820C gn 3 pht. Rt nng dung dch vo hp hoc l thu tinh c da bp ci, bt kn v lm lnh nhanh to mi trng chn khng.
Bp ci ti Trang 4

Ct phn li

Ct ming nh

p mui

Ln men

Rt nc ngm (Tu theo yu cu)

V bao b

Ngm da (Tu theo yu cu)

Lm lnh

Thanh trng

Da bp ci ti

Da bp ci ng hp

Hnh 2.1 S din t tin trnh ch bin da bp ci 2.1.3 Vi sinh vt trong qu trnh ln men
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2.1.3.1 Qu trnh ln men t nhin Khi bp ci ngm vo dung dch nc mui th nhng c tnh ca vi khun ln men bt u lin tip xy ra, u tin l nhm vi khun ln men d th hoc vi khun ng ln men ng th c sinh kh hoc khng sinh kh (Fleming cng cng s, 1988b). Leuconostoc mensenteroides l vi khun ln men lactic d th ln men u tin. L. mensenteroides vt tri trc tin bi v n hin din s lng cao (Mundt cng cng s, 1967; Mundt & Hammer, 1968; Schneider, 1988), pht trin sm giai on u, v c thi gian sinh si ny n ngn hn vi khun lactic khc trong nc p bp ci (Stamer cng cng s, 1971). Mc d vt tri nhanh, L. mensenteroides cht nhanh trong tin trnh ln men (Hnh 2.2), bi n nhy cm vi mi trng acid (McDonald cng cng s, 1990). L. mensenteroides c i km vi Lactobacillus brevis v Lactobacillus plantarum (Pederson & Albury, 1969). Lactobacillus cuvatus, Lactobacillus confusus, Lactobacillus sake, Enterococcus faecalis (trc y l Streptococcus faecalis, Lactobacillus lactis subsp. lactis (trc y l Streptococcus lactis), v Pediococcus pentosaceus (trc y l cerivisiae) c phn lp t ln men da bp ci hoc nhng sn phm c lin quan (Pederson & Albury, 1969; Puspito & Fleet, 1985; Yago cng cng s, 1985; Buchenhuskes cng cng s, 1986; Kandler & Weiss, 1986; Harris cng cng s, 1992a), nhng vai tr ca cc loi vi khun ny trong s ln men nhng ln sau cha r. Trong sut giai on sinh kh, vi khun ln men lactic d th s dng ng ca bp ci (ng glucose, fructose v sucrose) to thnh hn hp acid lactic, acid acetic, ethanol v CO2 (Fleming cng cng s, 1988b). Fructose c chuyn i to thnh mannitol, ng dng dng ny nh cht nhn in t. (Kandler, 1983). pH gim nhanh v kt hp thm mui NaCl, th vi sinh vt gram m b c ch. CO2 thay th khng kh v to mi trng ym kh, ngn nga oxy ho acid ascorbic v s sm mu t nhin ca da bp ci. CO2 cng kch thch s pht trin ca cc vi khun lactic khc. giai on cui ca qu trnh ln men, vi khun ln men lactic ng th Lb. Plantarum to thnh s lng ln acid lactic t hp cht hydro carbon cn st li, pH gim mnh. Cc tin trnh xy ra theo hng ng th s t c mt sn phm da bp ci n nh vi mi v v hng thm c trng. C hai tc ln men v tc vt tri ca mi ging loi b nh hng bi nng mui NaCl v nhit ln men. Nng mui tng th L. mensenteroides b km hm nhiu hn l Lactobacilli hoc pediococci (Pederson & Albury, 1969; Stamer cng cng s, 1971; Yildiz & Westhoff, 1981). Nu nng mui hoc nhit qu cao, th vi khun ln men lactic ng th pht trin nhanh, kt qu l qu trnh ln men lactic d th b rt ngn, t l acid lactic tng, v mu, mi v, cu trc km hn. (Pederson & Albury, 1953; Gangopadhyay & Mukherjee, 1971). P. pentosaceous v E. faecalis th c th pht trin khi nng mui NaCl hoc nhit trn mc bnh thng (Pederson & Albury, 1969). Pederson & Albury (1969) nhn nh rng da bp ci c th ln men tt nhit thp (7.50C). L. mensenteroides pht trin tt nhit thp hn vi khun lactic khc trong qu trnh ln men bnh thng. Kt qu, nhit thp acid tng l 0.4% (tnh theo acid lactic) c sn xut khong 10 ngy v 0.8 - 0.9% t hn mt thng. Tnh theo acid ny, cng vi s bo ho ca nhng ming bp ci nh trong nc mui vi CO2 sinh ra do vi khun, l nhng iu kin cn thit cung cp cho qu trnh bo qun. Lactobacillus v Pediococcus pht trin rt yu hoc khng c nhit thp.

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 nhit ln hn bng 32 0C, qu trnh ln men ch yu l vi khun ln men lactic ng th chim u th l Lb. Plantarum v Pediococcus pentosaceus (Pederson & Albury, 1969). Mi v v hng thm da bp ci xu i v bp ci b acid ho do hai ging vi khun ln men lactic ng th to thnh acid lactic vi hm lng ln. Da bp ci ln men nng cao nth d b sm mu do nn v hp nhanh sau qu trnh ln men. Khi sn xut 18 - 20 0C, acid tng cui cng khong 1.6 - 2.3% (tnh theo acid lactic) th sn phm (pH khong 3.5 hoc thp hn). T l acid khng bay hi trn acid bay hi (acid lactic / acid acetic) thng l 3:1 n 5:1 (Trail cng cng s, 1996). Qu trnh ln men c hon thnh 2 tun n 2 thng, ph thuc vo s lng vt cht c th ln men, nng mui, nhit , v s a thch ca ngi sn xut. 2.1.3.2. S nui cy men ging Vic cy men ging vo trong ch bin sn phm da bp ci trnh c vic da vo nhng vi khun t nhin c trong nguyn liu ti v c th lm cho tnh cht v gi tr ca sn phm tng cao. Tuy nhin s pht trin ca men cy vo th kh iu khin pht trin bnh thng v d xy ra qu trnh ln men t pht nu nguyn liu khng qua thanh trng, gy nh hng n cu trc. Trong mt th k qua thc hin nhiu phng cch p dng vic nui cy men ging vo da bp ci ln men. (Bo co v ghi nhn ca Pederson & Albury, 1969). Nm 1920 - 1930, Lc.lactis subsp. lactis (Streptococcus lactis trc y) c phn lp t cc sn phm sa th c lm men ging cho da bp ci (Brunkow cng cng s, 1925; Pederson, 1930a,b). Trong tt c cc trng hp khng cy men ging th sn phm da bp ci tt hoc tt hn. Tuy nhin, Pederson (1930a) tht bi vic phn lp Lc. Lactis subsp. lactis t bp ci ln men v ng ta xem cht lng da bp ci c ci tin l c cy men ging. Khi vi sinh vt ca da bp ci n nh hn th Lc. Lactis subsp. lactis c thay th bng vi khun lactic khc c tch ra trong qu trnh ln men. Trong nhiu trng hp, vic tim chng c lp hoc pha trn men ging L. mensenteroides, Lb. Brevis v Lb. Plantarum mt t, nu cht lng sn phm c ci tin hn so vi khng cy men ging (Keipper cng cng s, 1932; Lopez cng cng s, 1954; Narbors & Salunkhi, 1969; Yago cng cng s, 1985). Tuy nhin, i khi vic cy ging cng lm cho cht lng da bp ci tt hn (Pederson, 1930a,b; Buchenhuskes cng cng s, 1986). Tri li, i lc cng nhn thy bt li cho khi cy men ging vi Lb. Plantarum vo bp ci. (Pederson, 1930a,b; Pedrerson & Kelly, 1938; Buchenhuskes cng cng s, 1986). Stamer (1986) nh gi nc p bp ci ln men vi ging n thun chng vi khun lactic. Nc p ln men bi L. mensenteroides th c mi thm hi ho v thch th, trong lc ln men bng P. pentosaceus th th v ging nh gim. Lb. Plantarum v P. pentosaceus sn xut sn phm c v chn ngt, cu trc b khim khuyt, v nh gi khng th chp nhn. Harris cng cng s (1992a) nhn thy giai on u ca qu trnh ln men da bp ci th vi khun sinh ra c t v tm thy 2 ging Lc. Lactis subsp. lactis sn xut nisin. S dng m hnh nc p da bp ci ln men, nhiu tc gi chng minh rng lng nisin thch hp c th c ch c Lb. Plantarum nhy cm vi nisin, trong lc L. mensenteroides khng nisin pht trin t c mt t bo ln nht (xp x 5 x 10 8 cfu / ml) (Harris cng cng s, 1992b). Breidt cng cng s (1993) phn lp bin th ca ging L. mensenteroides khng c nisin mc cao v s dng nhng ging ny trong qu trnh ln men da bp ci cng vi nisin tinh khit mc cao (Breidt cng
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cng s, 1995). Nhn thy rng qu trnh ln men lactic d th pht trin v lm chm qu trnh ln men lactic ng th. Choi cng cng s, (1990) s dng nisin mc thp trong qu trnh ln men Kimchi lm gim bt t l sn phm acid trong qu trnh ln xy ra t nhin. Mc nisin cao v khng thm men ging khng nisin th s lng vi khun gram dng gim, vi khun gram m vt tri v gim h hng cho sn phm. L. mensenteroides sn xut D(-) acid lactic, l cht con ngi khng th chuyn ho c v khng cho php s dng vo sn phm cho la tui thiu nhi v tr nh n (WHO, 1974). c, ging Lb. bavaricus c dng sn xut dng L(+) acid lactic trong sn phm da bp ci, mt sn phm bn ch yu trong ca hng thc phm dinh dng (Lucke cng cng s, 1990). 2.1.4 Hao ht v h hng ca da bp ci Mt s vn h hng lm nh hng n da bp ci. Bao gm s bc mu (s oxy ho), gim tnh acid, c v v mi l (mi mc, mi men v mi i), c tnh nht, bp ci b mm, v da bp ci c vt hng, tt c nguyn nhn l do s pht trin hiu kh ca nm mc v nm men. Khc phc nhng vn ny bng cch m bo hot ng ym kh. Hao ht xy ra trong nhiu nm l da bp ci b mm hoc b nht. B mm l kt qu ca s to thnh dextran bi L. mensenteroides. Tuy nhin, dextran c sn sinh ra bi enzyme dextransucrase, c to thnh bi ng sucrose. Phn t ng sucrose c phn ct ra thnh ng fructose v glucose v glucose c thm vo lm cho chui dextran pht trin. Mc ng sucrose trong bp ci (Bng 2.3) c th thp hn mt vi loi polysaccharide ngoi bo khc, c th c sn xut bi mt vi sinh khc hn L. mensenteroides. S to thnh cht nht trong da bp ci cha bao gi c nghin cu hon ton. Bp ci c tnh nht do hot tnh ca pectinolytic l s hao ht lu di, nhng t ph bin. 2.2 DA CHUT MUI CHUA 2.2.1 Nguyn liu Da chut (Cucumis sativus) c trng hng ngn nm (Vaughn, 1985). Da chut cho nhiu dng mui chua trn th trng, chng c la chn theo hnh dng u n v cu trc vng chc. Da chut lm mui chua c thu hi lc cha chn. Da chut chn hon ton th khng ph hp bi chng qu ln, mu sc v hnh dng thay i, c nhiu ht gi, v phn ln l qu mm. Da chut c phn loi loi b tt c nhng da chut dp hoc gy, b hng hoc b dp nt. Vic phn loi bng my phn loi da vo ng knh da chut. Da chut mui chua c pht trin t Canada n Mexico v c nhiu nh ch bin chp nhn trn nhiu nm. Nu vn chuyn t ni xa, da chut ti c gi lnh hoc ngm trong nc mui phn phi ni xa hn. Hp cht hydro carbon c th ln men ch yu trong da chut mui chua l ng fructose v glucose (Bng 2.3). Acid malic c th hin din vi nng gy ra vn (xem bn di) khi chuyn i to thnh acid lactic v CO2 theo ng hng ln men malolactic. 2.2.2.Cng ngh ch bin Da chut mui chua c bo qun bng phng php ln men (40% sn phm M), phng php thanh trng (40%), phng php lm lnh (20%) (Fleming cng cng s, 1995a), nh pht ho hnh 2.3. 2.2.2.1 . Lm lnh vi th l Lm lnh vi th l mui chua c gii thiu Bc M nm 1960 (Fleming cng cng s, 1995a). Chng to cho da chut c mi v ti v cu trc rn chc.
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Da chut ti

Phn loi

Phn loi

Phn loi

Thng cha

Ra

Ra

Ngm mui

Ct hoc nguyn

Bao gi (Ct hoc nguyn) Rt dung dch

Mui t yu cu

Kim tra v chn

Cho ln men

V bao gi

y np

Phn loi

Rt dung dch

Lm lnh

Ct hoc nguyn

Lm kn

Kh mui

Thanh trng

Sn phm (v chua ngt) V bao gi

Rt dung dch

Rt dung dch

y np

Thanh trng

Vi sinh vt pht trin khng ng k trong qu trnh mui chua ny v c ngn chn bng cch thm chua n c nng chua bao gi ti bo qun nh Natrimui chua v Sn phm mui gim Mui thp, ho cht Lm lnh benzoat, 0 phng php lm lnh ( <5 C).
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Mui chua lm lnh khng c tnh acid c th c tm thy mt vi th trng c bit. Mui chua c thc hin bng cch ngm da chut trong nc mui NaCl 5% vi th l, ti. Nc mui da chut c tn tr v ln men 3 - 7 0C, ni m ln men lactic chm sn sinh acid tng (tnh theo acid lactic) khong 0.3 - 0.6% cui thng th 3. (Fleming cng cng s, 1995a). Ti nhit v nng mui thp th L. mensenteroides ng vai tr quan trng hn so vi da chut ln men khc. 2.2.2.2 Mui chua bao gi ti ( thanh trng) Mui chua c thanh trng c gii thiu M nm 1940, nghin cu ca Etchells & Jones (1942) v sau l Monroe cng cng s (1969). Da chut b acid ho vi gim n (0.5 - 0.6%) n pH xp x 3.7, men ging c thm vo, v nc gim c thanh trng n nhit tm l 740C trong 15 pht. Nng mui c th thay i t 0 - 3%, nng ng thay i theo tng loi mui chua. Nhiu ngi tiu th thch bao gi ti hn ln men chua bi mi v hi ho ca acid acetic (gim n). 2.2.2.3 Mui chua ln men Mui chua ln men, thnh thong c gi l sn phm mui hoc chnh cng l mui chua c ngm trong nc mui v tri qua qu trnh ln men lactic hon ton. Da chut phn loi theo kch thc, qu h hng v hoa c loi b. Hoa c mc nhim vi sinh vt cao (Etchells cng cng s, 1958) lm cho enzyme lm mm (polygalacturonase) m ca da chut. Da chut ti c cho vo trong b nc mui ln, m ca bn ln men, l kiu ln men t nhin. Nng mui ban u trong nc ngm khong 5 -8%. Qu trnh ln men thc hin nhit xung quanh khong 15 - 320C. Nc mui b acid ho vi acid acetic n pH = 4.5 hoc c CO 2 sinh ra v tc pht trin ca vi khun lactic nhanh. Nng CO2 cao l nhuyn nhn lm trng v phng ln ca m da chut (gi l s phng hp hoc s phnh ra). Costilow cng cng s (1977) pht trin phng php loi CO2 qu mc t nc ngm da chut. Nc da c acid ho vi acid acetic di pKa ca bicarbonat v khng kh (hn bnh thng) hoc N2 (ng k) to p lc ln thng ln men, lm cho vic loi CO2 c hiu qu. Vic loi b ny xy ra trong sut qu trnh ln men. Khi s dng thng gi, Kali sorbat (0.035%) hoc acid acetic (0.16%) c thm vo nc da ngn nga s pht trin ca nm mc v mm qu (Gates & Costilow, 1981; Potts & Fleming, 1982). Vi nng d mui va phi (5%) v nhit (20 - 27 0C) tin trnh ln men nhanh v hp cht hydro carbon ln men c khng cn, acid lactic hnh thnh nhiu. Ni chung, qu trnh ln men hon thnh 2 - 3 tun. Xp x 1.1% acid lactic hin din sau qu trnh ln men, vi pH cui cng khong 3.3 - 3.5. Mui chua sau khi ln men, c kh mui v ch bin vi nhiu sn phm ph gm ng v ngun mui chua, th l ch bin, v gia v. Mi v c th thay i, ph thuc vo lng ng, ging men v gia v thm vo. Acid lactic b loi b trong giai on kh mui v thng c thay th bng acid acetic. Mc d khng i hi, i khi mui chua ln men c thanh trng nhit thp tng thi gian bo qun. 2.2.3. Vi sinh vt trong qu trnh ln men 2.2.3.1. Qu trnh ln men t nhin Cc yu t nh hng n mt vi sinh vt trong ln men da chut bao gm nng mui v nhit nc ngm, nhng ngun dinh dng ln men, s lng v loi vi sinh vt hin din giai on bt u ln men. Tc ca qu trnh ln men ph thuc vo nng ca mui v nhit nc ngm. Trong sut giai on u ca qu trnh ln men, vi khun, nm mc v nm men c th b phn lp vi s lng ln. Bnh thng cui giai on u l 2 - 3 ngy v i
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khi di khong 7 ngy. Trong sut thi gian ny, vi khun lactic v qu trnh ln men v nm men oxy ho gia tng nhanh v vi sinh vt khng cn thit b gim thiu v cui cng bin mt ton b, l kt qu ca s gia tng nhanh chng ca acid tng v gi tr pH ca nc da gim (Fleming cng cng s, 1995a). Tin hnh gim pH xung 4.5 bng acid acetic th cc vi khun gram m b km hm v vi khun acid lactic pht trn mi v. Vi khun ln men lactic d th pht trin khng ng k trong sut qu trnh ln men da chut. Nhng vi khun nh L. mensenteroides thng b km hm bi nng v nhit nc da cao, v pH gim nhanh. Qu trnh ln men s khai hon thnh bi vi khun lactic P. pentosaceus, Lb. Brevis v Lb. Plantarum. Nhiu ging nm men c phn lp trong sut giai on u ca qu trnh ln men, mc d y l lnh vc khng th xc nh c tnh trit . Nm men ln men c phn lp gm Hansenula anomala, H. subpelliculosa, Saccharomyces bailli, S.delbruckii, S. rosei, Torulopsis holmli, T. lactis- condensii, v T. verstilis (Etchells cng cng s, 1961). Ging c tnh oxy ho c tm trong da chut ln men bao gm Candia krusel, Debaromyces hansenii, Pichia ohmeri, Rhodotorula spp. v Saccharomyces rousenii (Etchells & Bell, 1950). 2.2.3.2 Qu trnh ln men c iu khin Qu trnh ln men nng mui thp l rt cn thit. Trong lnh vc ch bin loi sn phm mui chua M, dung dch mui s dng c chc nng quan trng. Nng mui thp s gip cho sn phm da chut ln men t kt qu tt. Nng mui ca nc ngm trong thng thp c xut (Fleming cng cng s, 1988a). Dng nitrogen tinh khit vo thng ln men duy tr ym kh b mt nh thng. Bng cch thm calcium vo nc da, nng mui thp hn c th duy tr cu trc cng chc ca sn phm mui chua. (Buescher cng cng s, 1981; Fleming cng cng s, 1987; Guillou cng cng s, 1992). 2.2.3.3. Cch cy men ging S dng vic cy men ging vo da chut ln men c gii hn. Kt qu nghin cu ca Pederson & Albury (1961) cho thy cy vo sn phm da chut vi khun E. faecalis, L. mensenteroides, Lb. Brevis v P. pentosaceus. Lb. plantarum chu c acid cao v vy chng pht trin chim u th giai on sau ca qu trnh ln men da chut. Thc hin st trng da chut trc khi cy men ging, lm gim vi khun cnh tranh. Etchells cng cng s (1964) s dng phng php chn bng nc nng (66 800C) hoc tia bc x gamma (0.83 - 1.0 MRad) t c mc tiu ny. Etchells cng cng s (1973) chn Lb. plantarum bn acid cy vo da chut ra sch, ty trng v acid ho. Cht m natri acetat c s dng lng nh va phi pH gim dn v hp cht hydro carbon ln men hon ton. Thanh trng mi trng l rt cn thit bi v sn phm CO2 v bt hnh thnh. Vi khun ln men ng th to CO2 l vn u tin lm ri tr; tuy nhin, McFeeters cng cng s (1984) chng minh rng Lb. plantarum c th sn xut CO2 t s phn hu ca acid malic thnh acid lactic (qu trnh ln men malolactic). Da chut cha acid malic (0.2 - 0.3%) dn n nguyn nhn gy cho da chut b trng phng (McFeeters cng cng s (1982). Da vo biu chn mi trng, Breidt & Fleming (1992) nhn thy rng hu ht vi khun lactic trong da chut mui chua chim u th hn tin trnh ln men malolactic. Daseschel cng cng s (1984) tch ly dng malolactic m ca Lb. plantarum. Dng ny i khi c th c u th trong qu trnh ln men da chut v ngn chn CO2 sinh ra. Ln men da chut khng c NaCl ch t c kt qu trong phng th nghim nhng khi iu kin sn xut quy m thng mi th khng y c. t c iu ny, qu c chn (3 pht 770C) v ngm trong dung dch m calcium acetate cng
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vi cy Lb. Plantarum vo. Di iu kin quy m nh thng mi th da chut b trng v mt i s cng chc bi v khng kim sot c s nhim vi sinh vt sau khi chn. Sn phm cht dit khun lactic c phn lp t qu trnh ln men da chut (Fleming cng cng s, 1975; Daeschel & Klaenhammer, 1985; Daeschel cng cng s, 1990); tuy nhin, n ngy nay, chng s dng khng thnh cng nh nui cy men ging. 2.2.4. Hao ht v s h hng ca da chut mui chua Hu ht s hng hng ca da chut trong qu trnh ln men l do hot ng ca vi sinh vt, enzume gy h hng to thnh, dn n s mm m, clotridia hp th acid lactic pht trin, sn sinh mi l, hoc CO2 sinh ra khng ng k do tht qu b mo m hoc to thnh hnh vng mc b bc bn trong. Vic m b ph hoi v s hao ht cu trc hoc cng l do hot ng ca enzyme cellulolytic v enzyme pectinolytic, ni chung l mt mt v gi tr kinh t. S hao ht c bit nh h hng s phng ln hoc s phnh ra c th khc phc bng cch b sung thm mt s cht gia v khc. Qu trnh ln men th yu vi nm men c th xy ra khi ng cn li v vi khun lactic b km hm pH thp. Sau , khi pH v mui qu cao hoc acid ho thp, vi khun clostridia v propionic c th pht trin, dn n h hng. (Fleming cng cng s, 1989). Nhiu thng ln men tn tr ngoi tri vi b mt l ra v da chut b dm xung bi sc nng ca np y. Vic phi di nh nng mt tri (tia UV) lm gim s pht trin nm men oxy ho, nm mc v vi khun trn b mt. Khi thng ln men ym kh, c th ngn nga h hng bng cch duy tr pH thp v p ng y v s lng v cht lng mui. 2.3. LIU LN MEN liu c ch bin theo nhiu dng bao gm ng hp, ln men, trch ly du liu. liu thay i theo nhiu dng ng k, chng bn dng nguyn, tch ht, nhi (t, tiu, bt tiu, qu hnh nhn, vv), ct i, ct thnh bn, ct lt mng, nghin, hoc hn hp ca liu v n bch hoa ngm. liu ln men to thnh khi l dng ch bin khc a Trung Hi, trong lc sn xut ch yu M l liu chn ng hp ( khng ln men).Chi tit hn v tin trnh sn xut liu c nghin cu bi Cruess (1958), Vaughn (1985), Ferguson v cc cng s (1994) v Garrido- Fernandez cng cc cng s (1995). 2.3.1.Nguyn liu Nhng nm trc y vic sn xut oliu a Trung Hi nh cao (Ty Ban Nha, Th Nh K, , Moroco v Hy Lp) sn phm liu c tiu th vi s lng ln. Tiu bang ca California M sn xut ng hng th su (Garrido- Fernandez cng cc cng s, 1995). Mt s lng ln nhng loi liu c s dng cho sn xut liu thng mi. Trn th gii, ph bin nht l nhng loi Gordal (olea europaea regalis), Clemente (cng c bit nh Gordal Sevillano, Sevillano hoc Sevillana) v Manzalilla (olea europaea pomiformis). Loi Ascolano v Misson cng c s dng ti California ( Ferguson cng cc cng s, 1994). Mi loi c mt hnh dng, kch thc c th. liu l qu nh, tch b ht, c v ng mnh, hnh cu ko di. Phenolic glucoside oleuropein l hp cht ch yu to v ng ca qu liu. Oleuropein phi c loi b hoc b thu phn to sn phm n c. Loi b c bng cch ho tan trong nc hoc nc mui hoc thu phn trong dung dch NaOH long ( 1-2% ). Trong sut qu trnh chn, mu sc thay i t xanh sang tm hoc , v cui cng l mu en. Mu
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xanh ti ca liu to thnh bi s hin din ca chlorophyll; chlorophyll gim v anthocyan to thnh l kt qu ca s i mu. Tht qu chim 70-90% trng lng. ng glucose l hp cht hydro carbon c th ln men ch yu vi lng ng frutose v sucrose t hn. Hp cht phenolic to thnh xp x 1-3 % khi lng tht qu. Hu ht nhng hp cht ny l ortho- diphenol, n ng vai tr quan trng trong ln men nh tin trnh lm sm mu liu chn. Nng hp cht phenolic gim gn mt na trong qu trnh ln men chn t nhin. 2.3.2.Cng ngh ch bin Cc cng on ch bin c bn cho sn phm liu c ch ra trong hnh 2.4. Theo phng php ch bin truyn thng, nhng thng g trn c s dng cho qu trnh ln men. Hin nay, hu ht liu c ln men trong b btng c ph mt lp parafin hoc mng plastic pha trn hoc thng c nhum thp (sc cha l 10.00020.000 lt). 2.3.2.1.liu c x l dung dch kim trong nc mui (Loi liu chn en hoc chn xanh California) Sn xut chn en ng hp l phng php pht trin California nm 1990 (Ghi nhn ca Vaughn, 1985; Ferguson cng cc cng s, 1994). liu c mu xanh n mu anh o c thu hi vo nhng thng ma thu. Khi khng nhng thng cha ch bin tt c liu cng mt thi im, th liu c tn tr ym kh trong nc mui (phng php truyn thng), dung dch acid long (90% sn xut cng nghip s dng hin nay). Phng php truyn thng i hi nui n trong nc mui nng ban u l 5.0 7.5% NaCl, hoc 20 30 0 salometer. (Dung dch mui bo ho (26.5%) l 1000 salometer). Sau vi ngy, thm mui vo tng nng n 7.8 9%. i lc liu cn c l u nhau trn da ca chng lm gim s co nguyn sinh t bo qu trong sut qu trnh ngm nc mui. Qu trnh ln men lactic tnh, gip cho bo qun qu n khi ch bin. Nng acid lactic c th n 0.4 0.45% trong 4- 5 tun. Trong cng on tch nc mui kh iu khin loi ra v vy cn phi nm vng phng php. Dung dch acid cha 0.7% acid lactic, 1 % acid acetic, 0.3% natri benzoate v 0.3% kali sorbat c th s dng thay cho nc mui. Hin nay hn 90% khu cng nghip California s dng phng php acid ho tn tr liu. Ln men lactic khng xy ra di iu kin ny v liu tt hn mt cch ng k trong nc ngm tn tr qu. Phng php lm lnh th t ph bin bi chi ph tn tr c tin. S thm mu bn trong nhit thp hn 50C. Tn tr liu bng phng php iu khin kh quyn (2% O2) 5 hoc 7.50C c th gi c 9 12 tun, t gim cht lng (Ferguson cng cc cng s, 1994). Nu liu c thng kh trong sut qu trnh x l dung dch kim, hp cht phenolic t nhin kt hp vi oxy v polymer, to thnh sc t en. X l dung dch kim thch hp v liu l ra ngoi khng kh hoc lm thng kh nc v dung dch kim x l th mu en pht trin mt cch ng k.
Bp ci ti liu ti

Ra

Ra liu en ngm mui Trang 13 Ngm mui khng xgikimc Phn loi theo kch p mui Phnmen Ln loi Bao l

X l dung dch kim liu xanh x l Ra trong Ngmdch c dung men Phn loiPhngi kim Bao mui Ln loi theo kch

Bo qun

X l dung dch kim

Ra

Ngm mui

Phn loi Tch ht v nhi

Phn loi theo kch c

Tch ht v thi ming

Xp hp

Tit trng

liu x l trong dung dch kim (liu chn)

Hnh 2.4 Qui trnh sn xut liu vi ba phng php ch bin. (Trch t Garrido- Fernandez, 1995)

Hu ht mu hnh thnh nhanh pH khong 8.0 9.5. Khi thm calcium chloride nng thp (0.1- 0.5% ) th ci thin c bn mu. Dung dch kim c loi b bng cch thay nc mi ngy hai ln trong 3- 4 ngy v trn vi khng kh nn hoc khuy trn. Nc ra th c thay th bng nc mui long (0.8- 2.5% NaCl) cho hn 2- 4 ngy. liu phn loi sau xp vo ngn
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(ty ), vo hp v ch bin nhit. bn mu ca liu ln nht khi pH khong 7.0 7.5. Sau v bao gi, 2 2.5% dung dch nc mui c thm vo hp trc khi ch bin nhit trong 50 60 pht 116 1210C di p sut. Nm 1919, mt v ng c botulinum xy ra California lin quan n sn phm liu California ng hp. Vn ny lm thay i quan nim ch yu v ch x l cho sn phm ng hp c acid thp. T qu trnh x l nhit cho sn phm c p dng (Ferguson cng cng s, 1994) Sn xut liu chn xanh, liu ch xanh hoc vng rm c s dng v trnh khng kh nghim ngt trong sut qu trnh x l dung dch kim. Qu trnh ngm v v hp ging nh liu chn en. (Ferguson cng cng s, 1994). 2.3.2.2 liu xanh x l dung dch kim trong nc mui (Sevillan hoc Spanish- loi liu xanh) Qu c mu xanh n mu vng rm sau khi thu hi cho x l dung dch kim liu xanh. Nhng liu ny chn n im, ci qu c tch d dng t qu c ht nhng mu ca qu cha b sm mu. liu x l dung dch kim th hu ht cht ng b ph hu, ra sch loi b cht kim, v sau ngm nc mui ln men lactic hon ton hoc mt phn. liu tri qua qu trnh ln men hon ton c xem l Sevillian hoc Spanish- cc loi liu. liu ln men mt phn phi c bo qun bng cch thm acid hu c, tit trng, thm cht bo qun, hoc lm lnh. Dung dch kim pha long (1.3 3.5% ) c x l 12 210C khong 5-12 gi. Nng dung dch kim v thi gian thay i theo chn ca qu, nhit ch bin v cng ngh sn xut ring l. X l dung dch kim dng li trc khi n thm vo n ht qu (khong 2/3 n 3/4) gi li mt lng nh cht ng ca ci qu to hng v cho sn phm cui cng. Sau khi x l dung dch kim, liu c ra vi nc lnh mt hoc nhiu ln loi b cht kim. Trong qu trnh ra, c th thm vo dng HCl thc phm hoc acid mnh khc trung ho mt phn dung dch kim. liu x l dung dch kim c ngm mui 10 -13% NaCl. ngn nga s co nguyn sinh, ban u s dng nng mui thp ri thm mui mi ngy t mc mong mun. Tuy nhin, nng mui thp th d b h hng do s pht trin ca clostridia. Mui c thm vo trong sut qu trnh ln men duy tr nng mui 5 -6%. C th tng ln 7% hoc cao hn giai on cui qu trnh ln men trnh s pht trin ca vi sinh vt gy h hng. Nhit thch hp cho qu trnh ln men l 24 270C. cui qu trnh ln men pH khong 3.8- 4.4, 0.8- 1.2% acid tnh theo acid lactic (Ferguson cng cng s, 1994). Qu trnh ln men hon thnh trong 3- 4 tun hoc c th di hn, tu thuc vo nhit , nng mui v mt vi khun lactic ban u. liu c v bao gi bng l thu tinh (thng trong mt m hnh xc nh ), nhng l ny c rt dung dch nc mui xp x 7 % v c bt kn. Ln men liu xanh c th xp ngn v nhi t dng si thm nhiu trong nc mui. C hnh nh v nhng qu hnh nhn cng c nhi vo . liu nhi c th gi c nhiu tun trong 8% nc mui trc khi bao gi. Nu qu trnh ln men hon thnh, pH < 3.5 v nng NaCl > 5 % s thch hp bo qun. Phng php thanh trng c s dng khi qu trnh ln men khng hon ton hoc theo mong mun ca ngi ch bin thc phm. Mt vi ngi ch bin thc phm thanh trng 600C hoc s dng nc mui nng 80 820C. Thc t ngn chn s pht trin ca vi khun. 2.3.2.3. liu chn en t nhin khng x l (phong cch Hy Lp) liu phong cch Hy Lp, ph bin Hy Lp, Th Nh K v nhng nc Bc Phi, c lm t liu mui chua khi chng chn hon ton n mu tm hoc en nhng trc khi chng qu chn. p dng phng php khng x l dung dch kim v
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cht ng ch lc qua mt phn vo trong nc mui. Bi iu ny, chng c v qu v duy tr mt v hi hi ng. Chng c th bo qun bng phng php ln men trong nc mui, bng tit trng hoc than trng hoc thm vo cht bo qun. liu c cho vo thng cha v c che ph vi 6 10% nc mui. Qu trnh ln men xy ra bi h thng phi trn hp thnh ca coliform, nm men v lacto-bacillus spp. Acid tng cui cng ca nc mui thng thp hn 5 % vi pH khong 4.3 4.5 (Garrido- Fernandez cng cc cng s, 1995). Tuy nhin, di mt vi trng thi, liu chn en t nhin len men hon ton, th acid tng cao bng 0.8 1.0%. Qu trnh ln men to thnh acetic do nm men chu mui, nu nng mui 10%. Nu khng gi ym kh, mt lp nm mc, nm men v vi khun to thnh trn b mt ca nc ngm, nguyn nhn l do ng v acid gim, pH ca nc mui tng. S h hng c th t s pht trin ca clostridia, vi khun propionic, v c rh l vi sinh vt lm gim sulphate. 2.3.2.4 liu c p mui liu p mui kiu Hy Lp l p mui vi mui dng kh v tr nn kh hn khi tri qua qu trnh ln men. liu chn qu c ra sch trong 2 3 ngy. Sau chng c t trong thng vi lp mui xen k. Sn phm gi kh mt phn bi nng mui cao. 2.3.3.Vi sinh vt trong qu trnh ln men 2.3.3.1. Qu trnh ln men t nhin Cng nh qu trnh ln men da bp ci v da chut, qu trnh ln men ca liu da trn vi sinh vt c trong nguyn liu hoc nhng thng cha tn tr sn phm. Khng ging vi bp ci v da chut, nm men c vai tr quan trng trong qu trnh ln men liu. Nc mui cung cp mi trng tt cho vi khun lactic pht trin, vi ng glucose, fructose v maltose l ngun hp cht hydro carbon chnh c th ln men c. Phenol dit khun, bao gm thu phn hoc khng thu phn oleuropein, cng hin din mc cao v c th ng vai tr trong vic xc nh loi vi sinh vt chim u th trong qu trnh ln men. (Fleming cng cc cng s, 1969, 1973; Juven cng cc cng s, 1971; Ruiz- Barda cng cc cng s, 1991; Ciafardin cng cac cng s, 1994). Khng ging vi cc loi rau qu c th len men khc, liu x l dung dch kim lm gim bt s lng vi sinh vt lc u v pH ca giai on u qu trnh ln men tng khong 7.5- 8.5. Thm vo , x l dung dch kim v ra bng nc sch c th lc ng t liu ra ngoi lm gim cht dinh dng v gim hm lng ng. Vi khun lactic trong qu trnh ln men liu tm thy tng t nh trong qu trnh ln men rau qu khc. Vi khun sinh kh khng ng k so vi vi khun ln men lactic ng th. Ni chung qu trnh ln men liu xy ra ba giai on c th, i khi xem giai on h hng l giai on th t. (Garrido - Fernandez cng cng s, 1995). Mt s lng ln vi sinh vt hin din trong sut giai on u ca qu trnh ngm . Nhng vi sinh vt ny hin din tt trn qu nh l trong thng ln men. Phn ln vi sinh vt ny l vi khun gram m ( Pseudomanas spp., Flavobacterium spp., Aeromanas spp.) tt nh mt vi nm mc, v s b tiu dit di iu kin ym kh. Thnh vin ca nhm coliform - aerogenes ( Enterobacter cloacae, Citriobacter spp., Klebsiella aerogenes v Escherichia coli ) cng hin din di mc cao trong sut giai on u ca qu trnh ln men. Vi sinh vt c kh nng chim u th trong sut hai ngy u ca qu trnh ln men v dn dn bin mt trong sut giai on hai ca qu trnh ln men. Bi v nhm vi khun ny nhy cm vi pH thp, ni chung chng bin mt theo sau s gim pH bi s nhn ln ca vi khun lactic. giai on u s lng ca coliform v bacilli xut hin chm hn qu trnh ln men rau qu khc, gi c lu n 10 - 14 ngy. Vikhun lactic

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 giai on u ca qu trnh ln men ny l ging genus Pediococcus, Leuconostoc v Lactococcus. lm gim s pht trin ca vi sinh vt gram m trong giai on u, pH c th gim xp x 6.0 n khi c si CO2 hoc bng cch thm acetat, lactat, hoc dng HCl thc phm. Giai on hai ca qu trnh ln men c xc nh l qu trnh ln men bt u pH = 6.0 v tip tc pH = 4.5 vi khun gram m v c bn b bin mt. Giai on ny c th 10 - 15 ngy, c xc nh bi s pht trin ca lactobacilli ( tr lactobacillus plantarum v Lb. Delbrueckii) v nm men. S lng ln lactobacilli tiu biu t c 7 hoc 10 ngy sau khi ngm. Sau , s lng gi nguyn hoc gim chm khong 60 300 ngy ca qu trnh ln men. Vi k thut ln men hin i, ban u mt khng lactic b gim v Pediococcus, Leuconostoc v Lactococcus spp. Nhng vi sinh vt ny c th quan trong nu nng mui gim. Giai on ba ca qu trnh ln men bt u khi pH gim n 4.5 v kt thc khi hp cht hydro carbon ln men khng cn na. Vi khun c u th hn l Lb. Plantarum, vi mc thp ca Lb. Delbrueckii c tch ra. Thm vo , nm men ln men v oxy ho cng c tm thy mc cao va phi trong giai on ln men ny. Nm men ln men sn phm, ethanol, ethyl acetate, v acetaldehyde, l nhng cht xy dng mi cui cng ca sn phm. Thm vo , chng sinh mt lng vitamin, c th thc y s pht trin ca Lb. Plantarum (Ruiz- Barba & Jimenez- Diaz, 1995). Mt nm men thuc nhiu loi khc nhau. Hansenula anomala, Cadida Krusei, Saccharomyces chevalieri, Cadida parasilopsis v Hansenula subpelliculosa th c nh hng ln trong mt vi qu trnh ln men (Ghi nhn ca Garrido - Fernandez cng cng s, 1995), trong lc Marquina cng cng s (1992) cho rng tnh vt tri ca Pichia spp., tt nh Saccharomyces cerevisiae, Debaryomyces hansenii, Kluyveromyces lactis v Rhoddotorula mucilaginosa. Vic phn b nm men trong qu trnh ln men l cha r rng; tuy nhin, chng c vai tr to ra c tnh cm quan ring bitca sn phm liu ln men. Nm men oxy ho c th c hi bi chng s dng acid lactic v pH nng ln, dn n h hng. 2.3.3.1. Cch cy men ging Hu ht cc nh ch bin khng cy vo thng ln men vi ging men thun chng nhng s dng thm 1- 2 lt nc mui ln men t s ln men hot ho vo 200 lt nc mui ngm mu liu mi (Ferguson cng cng s, 1994). Hot tnh tiu biu ca nc ngm ny l pH = 4.0 vi hn 106 cfu / ml Lb. Plantarum, cho qu trnh ln men mi bt u tt. Nm 1930, Cruess ln u tin ngh s dng nc mui ngm ln men thng thng nh cht mi cho nc ngm liu mi. Nm 1937, ng ngh s dng Lb. Plantarum l mt men nui cy. Lb. Brevis, Lb, fermenti, Lb. Buchneri v L. mensenteroides cng c s dng nh s nui cy men mang tnh thc nghim; tuy nhin, s t trong chng chim u th hn vi sinh vt t nhin. Etchells cng cng s (1996) s dng nh s nui cy thun chng v pha trn ging tim chng Lb. Plantarum, Lb. Brevis, P. pentosaceus v L. mensenteroides sc nhit (740C trong 3 pht) v liu khng x l nhit vi nng mui thp (5 - 6%). S nui cy c th chim u th trong h thng sc nhit nhng khng trong liu khng x l nhit. Lb. Plantarum vt tri khi cy phi trn hai hoc ba ging . Lb. Plantarum LPCO10 nguyn thu c tch ra t liu xanh ln men v dn n sn xut ra hai vi khun: plantaricins S (pIS) v T (pTI) (Jimenez - Diaz cng cng
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s, 1993). S kt hp cc loi vi khun bao gm vi khun lactic, vi khun propionic v clostridia c kt qu c ch s pht trin ca vi khun gy h hng. Ruiz - Barba cng cng s (1994) ch rng ging ny sn xut cht dit khun, nhng khng bt ngun t vi khun tri du ca n, c th vt tri trong ln men liu - ngh rng sn xut vi khun c th sn xut trong s pht trin nui cy men ging cho liu. 2.3.4. Hao ht v h hng ca liu ln men Sau qu trnh ln men, liu c tn tr trong nc ngm. Do , iu chnh acid ho v nng mui thch hp l quan trng ngn nga h hng. c gi l giai on ln men th t xy ra trong qu trnh tn tr liu v khng ng k. liu c d tr trong nc ngm ln men vi mui thm vo xp x 8% n khi em bn. Nu liu khng c mui trong mt thi gian, hoc nu pH cui cng qu cao ( > 4.0), Propionibacterium spp. c th pht trin, to thnh acid acetic v acid propionic ti ph tn ca acid lactic. Nu giai on ny c tip tc, th c th thy c pH ca nc ngm gia tng nhanh, theo l s pht trin ca clostridia. c im h hng liu chn California l tht qu mm v da b trc. Vi khun gram m pectinolytic gm Enterobacter aerogenes v Aeromonas liquefaciens hu nh kt c lin quan n s h hng. Nu nng mui thiu hoc qu trnh ln men qu chm, vi khun coliform pht trin lm mm cu trc. Coliform v nm men c th l nguyn nhn sinh bt kh trong liu v to h hng fisheye. Thi gian ch ngm liu vo nc mui di c th gy hao ht sn phm. Gim s hao ht ny, to iu kin cho vi khun Lb. Plantarum pht trin, vi khun ny c di lp v liu. Nhng nm men mu hng Rhodotorula glutinis var. glutinis R. minuta var. minuta v R rubra c lin quan vi s lm mm liu.Chng sn xut enzyme polyglacturonase l nguyn nhn lm mm m. S pht trin ca nm men c th ngn chn bi duy tr trng thi ym kh (thng s dng) hoc dng tay loi b lp nm men (t s dng). H hng Zapatera ca liu c c im l mi khng mong mun cao trong qu ln men. Giai on u n c mi phomai nhng qua mt thi gian n tr thnh mi hi thi. H hng Zapatera xy ra khi ln men lactic dng li trc khi pH nc ngm gim xung 4.5 ( Kawatomari & Vaughn. 1956). Di pH ny, h hng Zapatera s khng xy ra. Trong giai on h hng, pH tng ln v nhiu hp cht c sn xut t s trao i cht ca acid lactic v acid acetic. Theo sau acid propionic thng l acid butyric, acid succinic, acid formic, acid valeric, caproic, v caprylic. Acid cyclohexan carboxylic c cho rng l 4- hydroxycyclohexan carboxylic acid, n xut hin trong nc ngm sau khi ln men lactic bnh thng ( Montano cng cng s, 1996). Propionibacterium v nhiu dng ca clostridium xut hin l nguyn nhn gy h hng Zapatera. H hng acid butyric xy ra th qu trnh ln men b tr hon ( Gililland & Vaughn, 1943). Mi v v ca butyric lm liu khng n c. S acid ho hoc vic cy men ging s ngn nga s h hng butyric.

PHN III. TRIN VNG TRONG TNG LAI

S gia tng chi ph s dng nc v nc thi trong ln men rau qu l vn cn thit cho vic nghin cu. Trng hp ca da chut v liu th c th ln men di

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nng mui thp hn phng php truyn thng s dng. Vn ny i hi iu khin mt s vi khun khng acid lactic v c th pht trin xa hn lnh vc cy men ging. Cy men ging vo iu khin cht lng sn phm ln men t nhin. Trong thi gian qua, nh gi s pht trin cht lng ca sn phm ch bin bng phng php cy men ging cha c ton din v do , kh xc nh chnh xc nguyn nhn cho s thnh cng v tht bi. ng dng di truyn phn t cho nghin cu tin b trong lnh vc ny (Breidt & Fleming, 1992; Harris cng cng s, 1992b; Fleming cng cng s, 1995a). Gy y, Passos cng cng s (1993 a, b, 1994) pht trin m hnh pht trin v sn xut acid lactic ca Lb. Plantarum trong da chut ln men. Nhng loi m hnh ny c th c p dng trong tng lai trong vic bo trc s tin b ca ln men rau qu. T quan im vi sinh vt hc, ln men rau qu l h thng xut sc cho nghin cu phi trn nui cy sinh thi hc

TI LIU THAM KHO

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1. Brain J.B.Wood. Microbiology of Fermented Foods, Vol.1. Blackie academic & Professional. 2. Lng, Nuyn c, 2002. Cng ngh vi sinh, tp 2. NXB i hc Quc gia Thnh ph HCM. 3. Lng, Nguyn c. Cng ngh vi sinh vt, tp 3- Thc phm ln men truyn thng. NXB Trng i hc K Thut Thnh ph HCM. 4. Phm, Lng c, 2002. Vi sinh vt hc v an ton vi sinh thc phm. NXB Nng nghip, H Ni.

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