Phân lập và sàng lọc vi khuẩn lactic có hoạt tính làm probiotic

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! 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