Science & Technology Development, Vol 11, No.

09 - 2008

THU NHN BACTERIOCIN BNG PHNG PHP LN MEN BI T BO LACTOCOCCUS LACTIC C NH TRN CHT MANG CELLULOSE VI KHUN (BC) V NG DNG TRONG BO QUN THT TI S CH TI THIU
Nguyn Thy Hng, Trn Th Tng An Trng i hc Bch khoa, HQG-HCM
(Bi nhn ngy 14 thng 06 nm 2007, han chnh sa cha ngy 19 thng 04 nm 2008)

TM TT: Trong bi bo ny, chng ti nghin cu vic c nh t bo vi khun Lactococcus lactis trn cht mang cellulose vi khun (Bacterial Cellulose- BC) ng dng ln men thu nhn bacteriocin v bo qun tht ti s ch ti thiu. Kt qu thu c nh sau: - Hiu qu s dng ch phm t bo vi khun c nh trn BC ln men thu nhn bacteriocin kh cao: c th ti s dng 9-10 ln m vn m bo v mt thi gian ln men, s lng v cht lng bacteriocin so vi i chng. - Bc u s dng mng mng cellulose vi khun (BC) hp ph bacteriocin bo qun tht ti s ch ti thiu: c th bo qun tht ti n 3 ngy bng mng BC hp ph dch bacteriocin 200 AU/m vn m bo cht lng tht, theo TCVN 7046:2002. - Kt qu thu c cng gp phn thm d 2 ng dng mi ca cellulose vi khun (BC): s dng BC lm cht mang trong k thut c nh t bo vi sinh vt v s dng mng mng BC lm mng bao thc phm. T kha: Bacteriocin, Bacterial Cellulose, Lactococcus lactis . 1. M U K thut c nh t bo ang c cp v quan tm nhiu, c bit trong cc lnh vc ln men sn xut cc sn phm trao i cht. T bo c nh c nhiu u im so vi vic s dng t bo t do nh: t bo sau khi c c nh c th s dng c nhiu ln, khng ln vo sn phm v c th ch ng ngng phn ng theo mun . Bacteriocin c sn sinh bi vi khun lactic l tc nhn sinh hc an ton trong bo qun thc phm thu ht s ch ca nhiu nh khoa hc. Bacteriocin l nhng phn t peptid do vi khun sinh tng hp c hot tnh km hm c hiu hay c ch mnh m s sinh trng v pht trin ca mt s vi khun gy hi trong thc phm [3, 4]. Mng mng cellulose vi khun (Bacterial Cellulose -BC) l mng polymer sinh hc. Mng BC c nghin cu s dng bo qun thc phm [1]. Trong bi bo ny, chng ti nghin cu vic c nh t bo vi khun Lactococcus lactis trn cht mang BC ng dng ln men thu nhn bacteriocin v ng dng mng BC hp ph bacteriocin bo qun tht ti s ch ti thiu. 2. NGUYN LIU V PHNG PHP 2.1 Nguyn liu - Chng vi khun: Lactococcus lactis spp lactis 1 (Lc.lactis) phn lp t pho mt. - Cht mang: Cellulose vi khun (Bacterial Cellulose- BC) [1]. - Mng bao thc phm: Mng mng cellulose vi khun (Bacterial Cellulose- BC) [1].

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Science & Technology Development, Vol 11, No.09 - 2008 2.2 Phng php nghin cu - Phng php c nh t bo: T bo c c nh trn cht mang cellulose vi khun (BC) bng phng php by- hp ph (gm 2 giai on: hp ph vi khun trn b mt BC v by tng sinh trn v trong cht mang BC). o Hot ho chng vi khun phn lp trong mi trng MT1 30 C, k kh, 6 gi. B sung cht mang BC v trng vo dch t bo hot ho. Qu trnh c nh trn cht mang BC s tri qua 2 giai on lin tip: Giai on hp ph: trong khong 30 pht u tin sau khi ngm BC trong dch t bo, BC s trng n v trng thi ban u, ng thi s hp ph t bo vo h thng si bn trong cng nh trn b mt bn ngoi. Giai on by tng sinh: k tip giai on hp ph. Cc t bo c hp ph giai on trn tip tc pht trin v tng sinh trn gi BC, khi tip tc iu kin ti u ca vi sinh vt c nh [1]. - nh lng vi khun c nh trn b mt mng BC: Qut mu i din bng giy thm Dp mu v trng- X l bng cellulose- nh lng bng phng php m gin tip [1,2]. - Ln men bi ch phm t bo c nh quy m phng th nghim trong fermenter (8 lt) thu nhn bacteriocin. - Xc nh hot tnh bacteriocin (AU/ml) theo phng php pha long hai ln lin tip (Schilliner v Rest) Xc nh pha long cao nht cho thy vng v khun v xc nh hot tnh bacteriocin (AU/ml) ca dch ln men:

AU/ml = DFi x (1/Vbactericin)

AU: n v hot tnh Dfi: pha long cao nht c vng c ch [5]. - Xc nh kh nng khng khun ca dch bacteriocin: bng phng php khuch tn trn thch. Phng php ny da trn kh nng i khng gia vi sinh vt kim nh v vi sinh vt ch th. Thc hin: Chng vi khun kim nh c nui cy trong 50 ml mi trng MT1 o 30oC, k kh, 24 gi. Ly tm 13000 vng/pht trong 20 pht 4 C, thu nhn dch ni. iu chnh pH dch ni n 6,5 bng NaOH 1N. S dng dch ni ny nh bacteriocin th, em kim tra kh nng i khng nh sau: + 15 ml mi trng vo a petri v trng. thch ng. + Tri 10l dch nui cy qua m chng ch th. + c l c ng knh 5 mm trn thch. + Bm 70 l dch bacteriocin th vo l thch. o + a 37 C, 12 gi. + Kim tra s to thnh vng v khun. o ng knh vng v khun [6]. 0 - iu kin bo qun tht heo nhit mt (15 C) - Thi im nh gi cc ch tiu cht lng tht ti sau 3 ngy bo qun. - nh gi cht lng tht heo ti theo TCVN 7046:2002.

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TP CH PHT TRIN KH&CN, TP 11, S 09 - 2008

3. KT QU V THO LUN 3.1. C nh t bo Lactococcus lactis trn cht mang cellulose vi khun (BC) bng phng php by- hp ph Cellulose vi khun - BC (do A.xylinum sn sinh ra) c ng dng trong nhiu lnh vc. BC c nhiu u th lm vt liu cht mang trong k thut c nh t bo [1]. i vi phng php by-hp ph trong cht mang BC, chng ti xt 3 yu t chnh nh hng n qu trnh c nh l: ch khuy o trong qu trnh hp ph, thi gian v nhit trong thi gian [1]. 3.1.1 nh hng ca ch khuy o trong qu trnh c nh bng phng php hp ph Phng php c nh t bo trn cht mang BC gm hai giai on: giai on hp ph v giai on by tng sinh. Giai on hp ph din ra trong 30 pht sau khi b sung cht mang BC v trng vo dch vi khun, lc ny BC s ht nc trong mi trng trng n v trng thi ban u. Khi nc i vo cu trc BC, n s mang c t bo vi khun i vo. Ti y, t bo s c h thng si cellulose trong cht mang BC gi li. ng thi trn b mt BC cng c t bo c hp ph. Tip theo l giai on by tng sinh, thi gian ko di ph thuc vo c im tng trng ca t bo c c nh. Cc t bo c nh s s dng c cht khuch tn t mi trng sinh trng v pht trin. Nh vy giai on hp ph c nh hng rt ln n hiu sut c nh t bo trn cht mang BC. ci thin giai on hp ph, ngoi vic n thun ngm BC trong dch vi khun (phng php A), chng ti cn kho st thm vic kt hp s dng cnh khuy (phng php B) v my lc (200 vng/ pht) (phng php C) trong qu trnh ny gia tng hiu sut hp ph. Kt qu ci thin mt t bo hp ph trn BC qua 3 phng php (phng php ngm, s dng cnh khuy v my lc-200 vng/ pht trong qu trnh hp ph) biu th biu 1. Hai phng php ci tin nhm gia tng kh nng hp ph t bo vi khun trn cht mang 8 u t c kt qu. Mt gia tng t 3,28x108 t bo/ g ln n 3,84x10 (phng php B) 8 v 3,86x10 t bo/ g (phng php C).
4 3.8 Mat o (10 8 te bao /ml) 3.6 3.4 3.2 3 2.8 A B Ph ng phap C

A: Phng php c in (ngm)

B: S dng cnh khuy

C: S dng my lc

Biu 1.Mt t bo vi khun c nh trn cht mang qua cc phng php hp ph.

Trong cc phng n ci tin, chng ti la chn s dng my lc 200 vng/ pht gia tng mt t bo hp ph ln cht mang trong thi gian BC trng n do tnh thun tin, d s dng v hn ch c kh nng nhim tp.

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3.1.2 nh hng ca thi gian trong qu trnh c nh bng phng php by-hp ph Thi gian chnh l thi gian vi khun tng sinh trn gi th BC, nht thm mt lng ln t bo vi khun trong mng li cellulose sau qu trnh hp ph [1,2].
7 Mt t bo x109 t bo/ g 6 5 4 3 2 1 0 1 2 3 T hi gian (ngy) 4 5

th 1. S bin i mt t bo vi khun theo thi gian trn cht mang BC

Kho st kh nng hp ph ca t bo vi khun Lc.lactis 1 trn gi th BC, nhn thy mt 9 t bo trn cht mang t gi tr cao sau 1 ngy c nh (3,46 x 10 t bo/g) v t gi tr 9 cc i vo ngy th 2 (6,41 x 10 t bo/g); cng ko di thi gian by tng sinh th mt t bo trn cht mang cng c xu hng gim ( th 1). Do dch vi khun dng c nh t bo Lc.lactis 1 c hot ho sau 12 gi nui cy. Thi im bt u a BC vo dch vi khun, t bo s c hp ph trn cht mang BC. Sau 24 gi, nhng t bo c nh ny s bt u i vo pha tng trng. Tip din ca qu trnh hp ph l qu trnh by tng sinh, t bo c nh s tip tc tng sinh v pht trin trn b mt gi th cng nh trong h si cellulose ca BC vi s c mt ca cc thnh phn dinh dng trong mi trng. Thi im 48 gi, vi khun i vo pha n nh v mt t bo t cc i (tng ng vi mt t bo vi khun hp ph trn gi th BC l cc i). Sau giai on ny, cc thnh phn dinh dng trong mi trng cng bt u cn dn, t bo i vo pha suy tn, v vy lng t bo c nh trn cht mang BC gim ng k. Vy, thi gian ti u c nh t bo Lc.lactis 1 trn cht mang BC l 2 ngy. 3.1.3. nh hng ca nhit n lng t bo c nh trong qu trnh by-hp ph
Mt t bo (x109 t bo/ g) 9 8 7 6 5 4 3 2 1 0 4 C 10 C 20 C 30 C Nhit 37 C 40 C

Biu 2. S bin i mt t bo vi khun c nh theo nhit trn cht mang BC

Nhit trong thi gian ca qu trnh by-hp ph chnh l nhit cn thit tng 0 0 sinh khi vi khun trn gi th BC. Vi 6 mc kho st nhit t 4 C n 40 C, lng t bo 0 9 pht trin trong cht mang BC cao nht khi 30 C t 7,67x10 t bo/ g (biu 2)

Nh vy, vi cc iu kin ti u c nh Lactococcus lactis kho st: hp ph bng 0 my lc, thi gian 2 ngy, nhit 30 C, lng t bo vi khun c nh trn cht mang t c nh sau: 9 - Mt trung bnh l: 7,67x10 /g. 4 2 - Mt vi khun mt ngoi cht mang l: 4,5x10 t bo/cm . 4 2 - Mt vi khun bn trong cht mang l: 3,8x10 t bo/cm . 3.1.4. ng dng ch phm vi khun Lactococcus lactis c nh trn cht mang BC ln men thu nhn bacteriocin
Hot tnh bac ter ioc in ( AU / m l) 1000 800 600 400 200 0 1 2 3 4 5 6 7 8 9 10 11 12 C S ln ti s dng ch phm c nh

Biu 3. Hot tnh bacteriocin ca cc ln ti s dng ch phm t bo c nh trn BC

Hot tnh bacteriocin trong 9 ln ln men ti s dng khong 800 AU/ml v khng c s khc bit so vi i chng khi ln men bng t bo t do. T ln ln men ti s dng th 10, hot tnh c xu hng gim (biu 3). Lng bacteriocin th thu c trong 10 ln ln men ti s dng khong 3,5 g/l, khng c s khc bit so vi i chng. T ln ti s dng th 11, lng bacteriocin gim nhanh cn 2,85 g/l. 9 9 9 Qu trnh ra tri t bo tng cng qua 10 ln ti s dng l (7,6x10 - 5,2x10 ) / 7,6x10 = 31% gp phn gii thch cho kt qu trn. Xt v c s lng v cht lng bacteriocin to thnh, c th ti s dng ch phm t bo c nh khong 9-10 ln. 3.2. ng dng bacteriocin bo qun tht ti s ch ti thiu 3.2.1. Kho st kh nng khng khun ca dch bacteriocin t Lc.lactis Vi mc ch s dng dch bacteriocin t chng Lc.lactis bo qun tht, chng ti tin hnh kho st ph khng khun ca chng Lc.lactis i vi mt s chng vi khun ch th, c bit l nhng vi khun gy bnh thng tm thy trong tht heo. T bng 1 cho thy dch bacteriocin t chng Lc.lactis c ph khng khun kh rng, c hot tnh c ch vi khun Gram dng v c ch mt s vi khun Gram m gy bnh nh Sal. typhimurium v E.coli.

Bng 1. Kh nng khng khun ca chng Lactococcus lactis

ng knh vng khng khun (mm) 11 10 12 0 14 15

Chng vi khun ch th

ng knh vng khng khun (mm) 16 12 15 12 13 11

Chng vi khun ch th

Bacillus subtilis Bacillus cereus Bacillus licheniformis 1 Bacillus licheniformis 2 Bacillus lacterlu Staphylococcus aureus

Salmonella typhimurium Escherichia coli Streptococcus sp Lactobacillus acidophilus Lactobacillus bulgaricus Lactobacillus plantarum

(a) Salmonella typhimurium

(b) Lactobacillus plantarum

Hnh 1. Kh nng khng khun ca dch bacteriocin ca chng Lc.lactis i vi mt s chng ch th

3.2.2. S dng mng mng BC hp ph bacteriocin bo qun tht ti s ch ti thiu Tht ti sau git m thng gim cht lng v c cht lng cm quan v cht lng vi sinh ch sau 12 gi nhit thng (bn cc ch) v sau 24 gi nhit mt (bn siu th). ng thi tht heo nhit thng li l mi trng l tng cho s sinh trng ca cc vi sinh vt gy h hng thc phm cng nh vi sinh vt gy bnh. Trong kho st ny, chng ti s dng mng mng BC kt hp bacteriocin bo qun tht ti s ch ti thiu. Bng 2. Cc phng php bo qun tht ti s ch ti thiu
Mu Ch tiu theo di Cht lng Mu sc Mi C0 C1 A B Sm mu, kh c trng C D E

Sm mu i nh

c trng

cm quan

Trng thi, cu trc

c trng t, hi nhy c trng

Mng BC trong sut khng nh hng trng thi cm quan. Khi bc lp mng, sn phm vn gi nguyn trng thi cu trc c trng

im chung (/20) TSVKHK E.coli S.aureus Salmonella pH Thi gian bo qun ti a (ngy)

20 4,5 x104 35

12 2,1 x106 50

13 1,8 x106 50

16 2,7 x105 35

18,5 2,4 x105 35

18,5 2,8 x105 35

18 6,9 x105 50

Vi sinh (CFU/g )

Khng pht hin 6,2 i chn g 5,0 1 5,0 1 6,0 3 6,0 3 6,0 3 6,0 3

(1) C0: i chng ban u: Tht heo ti ngay sau git m (2) C1: i chng th nghim: Tht heo ti theo thi gian bo qun (3) Mu A: S dng mng BC (4) Mu B: S dng dch bacteiocin c hot tnh bacteriocin 400 AU/ml (5) Mu C: S dng mng BC ngm trong dch bacteriocin c hot tnh 400 AU/ml (6) Mu D: S dng mng BC ngm trong dch bacteriocin c hot tnh 200 AU/ml (7) Mu E: S dng mng BC ngm trong dch bacteriocin c hot tnh 100 AU/ml Dch bacteriocin th c thu hi t ln men s dng chng Lc.lactis 1 bng phng php ly tm thu dch ni v trung ho v pH 6.0 bng CaCO3. Mng mng BC c nui cy v thu nhn, sau khi x l [1] c ngm 30 pht vo trong cc dch bacteriocin c hot tnh ln lt l 100, 200 v 400 AU/ml. Trong qu trnh 30 pht ngm, mng BC c trng n v hp ph dch bacteriocin vo mng. Mng mng BC hp ph bacteriocin c dng lm mng bc tht ti trong bo qun s ch ti thiu. Tht heo ti c thu nhn ngay sau khi git m v c chia lm 7 mu th nghim vi phng php bo qun khc nhau. Kt qu nh gi cht lng cc mu tht sau thi gian bo qun c trnh by trong bng 2. Xt v cht lng cm quan, da trn cc ch tiu v yu cu cm quan ca tht heo (theo TCVN 7046: 2002) chng ti c nhn xt sau: - Mu i chng th nghim C1, sau 1 ngy bo qun, mu sc, mi v trng thi cu trc khng cn duy tr c so vi mu i chng ban u C0. - Mu A, sau 1 ngy bo qun, tuy vn gi c mi c trng nhng bt u c du hiu i sau 1 ngy bo qun v khng gi c mu sc v trng thi cu trc so vi mu i chng ban u C0. - Mu B, sau 3 ngy bo qun, gi c mi v trng thi cu trc c trng so vi i chng ban u C0, tuy nhin b mt tht b kh. - Mu C, D, E, sau 3 ngy bo qun, gi c mu sc, mi v trng thi cu trc c trng so vi i chng ban u. So snh im chung nh gi v cht lng cm quan cho cc mu kho st, nhn thy 3 mu C, D, E s dng bacteriocin c nh trn BC bo qun tht ti u c im cao (>18/20 im) so vi cc mu cn li. V vy, da vo cht lng cm quan cc mu C, D v E c chn.

Xt v ch tiu vi sinh v pH, da trn qui nh k thut ca tht ti v ch tiu vi sinh vt v ch tiu pH theo tiu chun Vit Nam (theo TCVN 7046:2002), chng ti nhn thy: - Mu C1 c tng s vi sinh vt hiu kh v E.coli cao hn hn so vi mu C0 v khng o t gii hn cho php. Do trong thi gian bo qun 24 gi 15 C, nhiu vi sinh vt hiu kh vn c kh nng tn ti nn vi iu kin mi trng tht giu dinh dng, chng tng trng nhanh chng. - Mu A c tng s vi sinh vt hiu kh v E.coli cao hn hn so vi mu i chng C0 v khng t gii hn cho php. Tuy nhin, tng s vi sinh vt hiu kh mu A li thp hn mu C1. Do mu A c bc bng mt lp mng mng BC nn gim bt lng khng kh xung quanh mu, v vy cn tr s tng trng ca mt s loi vi sinh vt hiu kh. - c bn mu bo qun tht B, C, D v E c x l vi bacteriocin, tng s vi sinh vt hiu kh cng nh lng E.coli thp hn so vi mu C1 v A rt nhiu (bng 2). Do bacteriocin ca chng Lc.lactis 1 c kh nng khng mt s vi khun hiu kh, do vi s hin din ca bacteriocin c ch s sinh trng v pht trin ca nhiu loi vi khun hiu kh c trong mu tht. Xt v cht lng vi sinh, c hai ch tiu tng s vi khun hiu kh v E.coli, cc mu B, C, D, E u t ch tiu theo TCVN 7046:2002 v QBYT 867. - So snh gia hai mu B v C, lng E.coli l tng ng nhng tng s vi sinh vt hiu kh mu B cao hn mu C (bng 2). Do bacteriocin mu B trng thi t do, ban u chng ph trn b mt tht, sau dn dn s thm vo khi tht. Di tc ng ca cc enzyme protease c trong khi tht, nhng phn t bacteriocin vi bn cht l protein s b phn hu dn. Tri li, bacteriocin mu C, c c nh bn trong cng nh trn b mt mng BC nn rt t chu tc ng ca protease. V vy, tuy c 2 mu u c x l vi dch bacteriocin c cng hot tnh (400 AU/ml) nhng hiu qu khng khun ca mu C li cao hn mu B. - Gia 3 mu C, D, E u c bo qun bng mng BC hp ph bacteriocin, nhng hot tnh bacteriocin cng gim, kh nng khng khun cng gim, v vy hiu qu bo qun cng gim, iu ny ph hp vi l thuyt. So vi TCVN 7046:2002, ch tiu tng s vi khun hiu kh ca cc mu C, D, E u nm trong gii hn cho php. So snh gia hai mu C v B, ch tiu ny khng c khc bit nhng khi so snh hai mu C, D v mu E th tng s vi khun hiu kh mu E cao hn 2,8 ln so vi mu C v 2,5 ln so vi mu D. V vy mu E b loi. Gia 2 mu C v D, khng c s khc bit, chng ti chn mu D do mu D s dng lng bacteriocin thp hn mu C nn thch hp hn cho mc ch bo qun thc phm nhm m bo ti a an ton cho ngi tiu dng v hiu qu kinh t. Ngoi ra, khng c pht hin v S.aureus v Salmonella trong 7 mu th nghim. i vi pH, mu C1 v A u khng t ch tiu. pH gim nhanh (5,0) so vi mu i chng ban u C0 (6,2). 4 mu B, C, D, E, pH khng khc bit nhiu so vi i chng (6,0) v trong gii hn cho php . Tm li, qua kho st cc mu tht ti s ch ti thiu c bo qun bng cc phng php khc nhau, chng ti nhn thy: Da vo cht lng cm quan: cc mu c chn l C, D v E. Da vo cht lng vi sinh: cc mu c chn l C v D. Do cc mu ti u cho bo qun tht ti s ch ti thiu l C v D. Gia hai mu C v D, chng ti chn phng php bo qun mu D v ch cn s dng dch bacteriocin c hot tnh 200 AU/ml. Cc kt qu phn tch trn y cho thy: mng mng BC ch c tc dng gia tng cht lng cm quan sn phm; bacteriocin c tc dng ci thin cht lng vi sinh ca sn phm. Kt hp mng mng BC hp ph bacteriocin c tc dng va tng cht lng cm quan v cht lng vi sinh sn phm tht ti s ch ti thiu.

Vy, dng mng mng BC (bacterial cellulose) hp ph dch bacteriocin c hot tnh 200 AU/ml c th bo qun tht ti s ch ti thiu 3 ngy so vi i chng ch bo qun c 1 ngy, vn m bo theo TCVN 7046: 2002. 4. KT LUN - Hiu qu s dng ch phm t bo Lc.lactis 1 c nh trn cht mang BC trong ln men thu nhn bacteriocin kh cao: c th ti s dng ch phm 9-10 ln m vn m bo v mt thi gian ln men, s lng v cht lng bacteriocin so vi i chng. - Bc u s dng mng mng BC hp ph bacteriocin bo qun tht ti s ch ti thiu: c th bo qun tht ti n 3 ngy bng mng BC hp ph dch th bacteriocin 200 AU/m vn m bo cht lng tht theo TCVN 7046:2002. - Kt qu thu c cng gp phn thm d 2 ng dng mi ca cellulose vi khun (BC): s dng BC lm cht mang trong k thut c nh t bo vi sinh vt v s dng mng mng BC lm mng bao thc phm.

IMMOBILIZING LACTOCOCCUS LACTIC CELL AND APPLICATING BACTERIAL CELLULOSE MEMBRANE TREATED WITH BACTERIOCIN TO PRESERVING PORK
Nguyen Thuy Huong, Tran Thi Tuong An University of Technology, VNU-HCM ABSTRACT: In this review, we investigate the immobilization of Lactococcus lactis spp lactis 1 in bacterial cellulose and application for producing bacteriocin. The results are given as following: - BC entirely satisfied the conditions required for immobilizing bacterial cell technology. - The effect of using Lactococcus lactis spp lactis 1 cell, immobilized in BC, for producing bacteriocin was hight: it could be reused about 9-10 times, the quantity and quality of bacteriocin remained rather good against control experiments. - Application of this bacteriocin, we us it to preseving pork. In turn, bacterial cellulose membrane treated with bacteriocin solutions which enhibit bacteriocin activity about 200 o AU/ml. Using this preparation to enveloping pork samples and maintaining at 10-15 C. After three days, comparation of sample and control experiment has many differences in ceptor quality. TI LIU THAM KHO [1]. Nguyn Thy Hng (2006). Tuyn chn v ci thin cc chng Acetobacter xylinum to cellulose vi khun sn xut v ng dng quy m pilot. Lun n Tin s Sinh hc. Trng i hc Khoa hc T nhin TP.HCM. [2]. Krystynowicz A, Czaja W (2002) Factors affecting the yield and properties of bacterial cellulose. Industrial Microbiology and Biotechnolody 29:189-195.

[3]. Lee K. Y. (2000) Survival of Bifidobacterium longum immobilized in calcium alginate beads in simulated gastric juices and bile salt solution. Applied and Environmental Microbiology 66: 869-873. [4]. Sheu T.Y. (1999) Microentrapment of Lactobacili in calcium alginate gel. Food science 54: 557-561. [5]. S.T.Ogunbanwo et al (2003) Characterization of bacteriocin produced by Lactobacillus plantarum F1 and Lactobacillus brevis OG1, African Journal of Biotechnology 2: 219-227. [6]. Shah N.P. et al (2000) Probiotic bacteria. Food science 8: 563-572.