22 - U 37°C/ 48 gia/ 5% CO2 quan sat kha nang phan giai casein sau hai nuoi cfty 2.3 7.3 Lipase •!• Nguyen t&c M vi khuftn c6 kha nang sinh enzym lipase, phan giai dfiu thµc v�t va cac hgp chftt beo khac Trong thfr nghi?m lipase, moi truong nuoi cfty c6 CaCli va Tween 80 N€u vi khuftn c6 kha nang ti€t enzym lipase, thi lipase se thuy phan Tween 80 tc,10 cac acid beo tg do, du&i SlJ c6 m�t cua ion Ca2+, tua calcium oleate dUQ'C tc,10 thanh, tc,10 nen mng tren moi truong MRS, 37°C, 5% CO2 sau 48 gia D8 tach chiet bacteriocin tho ta tien hanh ly tam 13.000 vong/phut, 20 phut, & 4°C Dich n6i thu duqc di�u chinh v� pH b�ng NaOH lM d8 tranh kha nang khang khufin cua acid vo ca Dich n6i sau d6 dugc tua b�ng dung dich ammonium sulphate (40% bao hoa) Khufty dung dich 2h/ & 4°C, sau d6 ly tam 20.000 vong/phut, lh, & 4°C Chdt tua duqc huy�n phu lc,1i vao 25 mL dung dich d?m potassium phosphate (pH = 7) Dich khufin chi thi duqc trai tr�n moi t�ang phu hqp va,d\lC nhfrng �i€ng c6 duong kinh mm Barn 70 µL dich noi da chuan bi vao moi gieng va u & C 30 phut d8 cac chdt-long khu€ch tan d€ dang thc,1ch Cuf>i cung cac dia duqc u a tung di�u ki?n phu hgp cho tung vi khufin chi thi Kha nang khang klmfin cua dich bacteriocin tho duqc xac dinh b�ng each duong kinh vong VO khufin quanh m6i gi€ng Trong nghien cuu chung toi khao sat kha nang khang khufin tren cac chung gay b?nh sau: Escherichia coli ATCC 25922, Salmonella spp, Bacillus cereus, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa, Vibrio cholera, Vibrio parahaemolyticus, Streptococcusfeacalis, Streptococcus tieu huy€t � nh6m A 29 CHUONG3.KETQUAVABAN LUAN 33 I ! l Hinh 3.7 Dinh tinh acid lactic chting K15 bling thu6c thii' Unphenment - Ong 1: Thu6c thu unphenment + nuoc cfit Ong 2: Thu6c thfr unphenmnet + MT MRS chua nuoi ciy Ong 3: Thu6c thu unphenment + acid lactic 2% Ong 4: Thu6c thu unphenment + djch nuoi c:iy vi khuAn 3.2 KHA NANG SINH ENZYM NGOAI BAO VA HOAT TiNH KHANG , KHUAN y o Trong cac tieu chi tu �n chon cac chung lam probiotic, m t nhung ho�t tinh tieu biSu cua probiotic la kha nang khang khuAn � Thu nghi�m ho�t tinh khang khuAn voi tac nhan b�nh hay kha nang sinh enzym la m(>t cac buac cua yeu c§u tuyBn chon probiotic, do tuy�n chon cac chung vi sinh v�t vira co ho�t tinh khang khmin d6ng thoi co kha nang sinh enzym ngo�i bao la tieu chi quan d� co the 1.'.mg dµng lam probiotic ma chung toi c§n ti�n hanh.[8, 32, 34] �Kha nang sinh enzym ngo�i bao va kha nang khang vi sinh v�t gay b�nh dm,Tc trinh bay iJ bang 3.1 I I 1, \ ! Bang 3.1 K�t qua khao sat kha nang sinh enzym ngo�i bao va ho�t tfnh khang khufin = ,- '"" \ i)rJiliHlLJ.F!Bi _ _' -�-·; ; -, -�·-! Hii , I -, ; - � rifitl-H -, m l -F-J , } f -1r;:rfoi/ !: � _ i £ ; -: � " i:' -· ' _ _ fil ' o � : µ;1: ; -( J ; l:%�(-1 li il ll f,,1ir1Jn� , I f , - , , - � �- · ��·- �- � , �- l; 4.1 CJ SCJ» ' -� a -� 9� , ,,,,,, '" '"'J·a., , ,, � < I l Il ?' � [1}iff''' : ] • !_fl�? ·- ' ] , -' i ffil] ''' L �™' _ , _.,.1I r II�,-·-·· + '''1 I · , J [(�{� ··-·-1 - ; -·-�J [ mr; )J! "' • ';i' ; , " ·1 + l { '.i ]l r· [ ·'· ·-·"· · -·· 'flf:;·· VJ - + PI�ff1i�:fi ,�·�::·�··�:·J·] [ rl.1l" _, ' "'·.· ·_ · ···-· ··1 ri U�Jl ,;d ,,_ ut�t�i":iqfj ' "''_"".J""l • ,• - + + + + + + - - 0±0 0±0 0±0 0±0 15 ± 0,67 16,2 ± 0,39 0±0 0±0 0±0 0±0 0±0 0±0 0±0 0±0 O::!:O 0±0 ·• 0±0 13±2,67 ()±0 0±0 0±0 0±0 0±0 10,3 ±0,22 0±0 0±0 0±0 6:fo· 0±0 0±0 0±0 0±0 � � st::s s � ,, 0±0 0±0 16±0,67 0±0 0±0 ±0 13,3 ± 0,22 n ±0,61 10±0,67 E c '5• � ,, ' '::::.: 0±0 0±0 0±0 0±0 0±0 0±0 14,8±0,39 o':ho 0±0 0±0 0±0 0±0 0±0 ·, 0:1::0 ± () 0±0 i:.,., 0±0 • ··· 0±0 10,3±0,22 0±0 0±0 0±0 0±0 9±0,17 0±0 0±0 t,:) '' ·.::: 0±0 0±0 15,3 ± 0,22 0±0 � � 12,4± o·,1s 'o:fo ()±0 :.:::I,} � � (}:I::() 0±0 0±0 � E �� � 0±0 0±0 0±0 � :::: ' '- • 0±0 O:i:0 0±0 ·t :�: : . -. ::::-:-::- � , 15,9±0,11 17±0,67 0±0 0±0 O±{l, i3,4:±: 0,19 s � :::i � �±0 0±0 0:±:() :: � 14,9±0;49 0±0 :"':"'��� :,: -.- 0±0 15,3 ± 0,22 ··· 16,5±0,17 -··· c;.,j 0±0 0±0 12,2±2,06 17,4±0,44 Ii±o,67 + ' 0±0 0±0 � ' ' Q.; ""'4,,,1 s t.tl 17±0,67 12±0,67 � ·;:, :.::: 0±0 12 ± 0,67 i j + 0±0 L· �· � -·-i � +,.;j,:I 0±0 + .l"' [ t11�w� _····-· - J-·� -rT Xr.J1 + L: : ·· [i'hh' - ·i ri'1,� I,} s.; � f :, u '!:cl + l"f·-fi·-··1) · """J.,, · f� 12 ± 0,67 0±0 0±0 0±0 ·; :1:·o 0±0 0:±:0 0±0 o :±: o· () :I: 11 ±0,67 0:±:0 0±0 (±0 11 :±: o,°7 0±0 0±() 0±0 S"i; & �� 0±0 0±0 0±0 0±0 [rti.J±L� 0±0 0±0 ' 0±0 0±0 0:f:O ', ()±0 0::1:0 0±0 ) 0±0 0±0 0±0 0±0 0±0 () ±0 0±0 0±0 :J us±o,'6� 0±0 0±0 0±0 :·o±o 0±0 () :±:() Bang 3.11 Tom tilt k�t qua thii' nghiem ciia 19 cltiing dmfc sang IQC ·;:,,,;t,lhr, ,m"fw � j K5 R4 S1 I m,)i1i mmi itJ!lnil!:: lUimtn 7/9 clfung Kefir Rau qua 7/9chung lenmen 5/9chung Suam� � I Suam� I 719chung I Suam� I 7/9chung I 1{6 I Kefir � S7 K7 R6 I Suam� I Kefir K9 Rau qua lenmen I Suachua qua I Rau lenmen I Kefir R14 I Kefir Yl R9 Y7 �� Sfrachua Sfram� I 5/9cluing 4/9chung 4/9chung it,,v;,:q,1 !&rti iHl,I\ pur,1c·,1.t,l L v11rornrd �uh,p wlam1s L p11rnlime11t,id1ir /,.J)tlrrlphlflll/tll/11 / pr/1/0SI/S I prm!t•n< L.1>lm1wm111 L rJ1111111wsm I wkf!i subsp sr!J -i L 1,1At'I soll,p rnmo111.1 I :;:me '€.u )!, =- ==· Ly'i·DA�p L)',;·DAsp mDpm mDpm LY�·DAsp mDpm" Lys-DAsp Ly�·DNp L)S·DAsp Lys-[)Asp L�':\·DAsp L��·DAsp NO ND mDpm Lys-l)Asp L�':\·DA�p Lis-DAsp Ly,al)Asp L}':\•PA�p Ly,-DA1p" Lys·l)Asp L)':\•DAsp Lys-DAsp Lys-DA�1>'' mDpm mDpm Ly,·DAsp mDpm Ly$·DAsp '-Y"·DA,p Lys·DAsp Ly,-DAsp c " �- B u Sl OS 3.5-36.5 3'J 42 43-44 '.l6Jl±3 3fr37 4� 45 45-47 45 45 42-44 42- 44 " - -Eu · ,,o � ; � E J § 02, , m !.(+) -i-i- L(ti) +!+!- DI 4-l- $! � I I.+ !)I I 15i45 -ft: "" o,;: ;,,, � d ·+ +l+ +i- + -tl- n +f- Pl "2 d Nl1 ND ll(I.) IJU'l.(!>) E +l- +I- - - - - d f'\l) NU -I- ND 41-41.7 41-43 33-35 35-Jt> l(D) +lND 37 33-35 35.37 :l5 43-U 45-47 45-47 37-3� 44,4,'i 4(1-47 49-.13 4-l-4(i 45-17 42-14 42 44 48-W PL l)I t>l m I)], !> Ill L 1; L ND' l pt m l UL I, m/1.(n) IHit(n) I.(U) Symb()ls and abbrcvia1inns; refer to footnote in Tabk 'fllcuUativci)' hct�rofcrm 111mui,·c 1'\V Et lamnn:-s unpul>Ji�hei.l results 'Str;iin� formerly dcsiinntcd /• + + - ND t - + + + t ND + + + + + t- - - - + w - u d - + d - - + d - -� - + - t - - - :I' - - - + + d d + + ti - + + + + + + + + + + - - - I t •· - + + + d + + - + + d + + + t ::2 - - - - - - - +