Tgp chi Cong nghe Smh hoc Ud): 145-152,2013
K H A O S A T M O T S O D A C T I N H C U A C H U N G LACTOBACILLUS PLASTARUM DC2 PH.A.N LAP T U S A N P H A M D I T A CAI T A l T H A N H P H O H U E , V I E T N A M
Do Thj Bich Thiiy', Phan Thi Be', Triin Thi Ai Luy^n' 'Trudng Dai hoc Nong Ldm, Dai Hpc Hue 'Truang Cao ddng nghe Du lich Hue
TOM T A T
Chung DC2 phan lap tir san pham dira cai da dugc dmh danh so bd dya tren cae dac diem smh hda va sinh ly cho thay thupc chl Lactobacillus. Dinh danh den loai bang phirong phap giai trinh tu gen 16S rDNA cho thay chiing nay Ihudc Ioai Lactobacillus ptanlarum Ham Iirong lactic acid trong mdi tmdng ducrc xac dinh bang HPLC sau 72 h nudi cay chiing nay trong mdi trudng MRS Idng c6 bd sung glucose (20 g/I) a 37°C dat 23,41 g/l Mdt so tinh chat the hien tiem nang probiotic ciia chiing nay cflng dugc khao sat Ket qua cho thay rang chiing nay co khd nang chiu acid tot Phan tram sd te bao sdng sdt so vdi ban dau sau khi u d pH 2 Irong I h, 2 h va 3 h lan lirot la 85,56 %, 74,63% va 7L47 %. Ben canh do, kha nang chiu muoi mat va bam dinh ciia nd ciing kha cao; sau 4 h ii trong dung dich chiia 0,3% muoi mat, ADD do d 600 nm dat 0,377. Kha nang tu ket dinh va ddng ket dinh \in Staphylococcus aureus la 38,57% va 24,23%, kha nang bam dinh vdi cac dung moi xylene, ethyl acetate va chloroform Ian lugt 14 61,46%, 43,27% va 74,81%
Tirkhda: Dinh danh Laciobacittiispic ten men lalic, phdn Idp, probio
DAT VAN DE
Vi khuSn lactic tdn tai nhieu Irong cac san pham len men chua a Hue nhu tdm chua, nem chua, dua chua...Dac biet he vi khuan lactic duoc phan lap tis cac san pham len men chua rau qua thich nghi dugc vdi didu kien dinh dudng ngheo, do dd cd kha nang tigm an nhirng chiing vi khuan lactic phat trien manh khi dugc cung cap day du dinh duong Ngudi ta thudng sir dung vi khuan lactic de san xuat lactic d quy md cdng nghiep Ben canh do, he vi sinh vat nay cdn duac ung dung frong san xuat thirc an gia siic, san xuit cac ch^ pham probiotic, san xuat cac san ph4m thuc pham chiic nang . Chinh vi vay, vi khuin lactic la mgt trong nhimg nhdm vi sinh vat cong nghiep quan trgng nhat.
Hi?n nay, cac nghien cuu ve vi khuan lactic da va dang dugc tridn khai rgng rai tren the gidi. Nair va Surendran (2005) da phan lap va xac dinh dac tinh Iy hda mdt sd vi khuSn lactic phan lap tir ca ya ldm Park va Oh (2007) da tuygn chgn vi khuan lactic tir sira chua. Cac ddng vi khuan sinh lactic acid cao Cling da dugc nghien ciru tuyen chgn (Kaloyan el al. 2008; Cock, Stouvenel, 2006); Mgt sd cdng trinh khac da tdi uu hda kha nang len men lactic, len men lactic vdi ngudn nguyen lieu re tien (Ohkouchi, Inoue, 2006, Zhengdong el al, 2009) Cac nghien ciru tuygn chgn. khai thac ti^m nang probiotic duoc
tnen khai rdng rai tren the gidi. Nghien ciru kha nang smh bacteriocin va kha nang irc che vi khuan gay benh ciia vi khuan lactic cho thay phan ldn cac chiing VI khuan lactic deu Iao ra duac bacteriocin irc che Lisleiia, Staphylococcus aureus (Rhys el al.
2008, Renata et al, 2004) Ben canh do, Kos va ddng tac gia (2003) da lien hanh n ^ i e n euu kha nang tu ket dinh, ddng kdt dinh va bam dinh dung mdi, bam dinh bieu md rudt ga cda chiing Lb acidophilliis M92, Provencio va ddng tac gia (2009) da nghien cim kha nang bam dinh cua Lb casei Kha nang thuy phan mudi mat va loai bd cholesterol trong dieu kien in vitro va in vivo ciia cac chung cd tiem nang probiotic da duac Liong va Shah (2005); va Maragkoudakis va ddng tac gia(2004)cdng bd
d Viet Nam, da bat dau cd mdt so nghien cim ve VI khuan lactic nhung ehua duac phong phii, Nguyen Thd Trang va Trin Dinh MSn (2008) nghien ciru mdt sd dac diem phan loai ciia hai chiing vi khuan lactic HNl I va HN34 sinh tdng hop L(-i-)-Lactic Mai Dam Linh va cdng su (2008) phan lap va tuyen chgn duac cac ddng hinh que mang nhieu dae diem ciia chi Lactobacillus va da lien hanh sang lgc kha nang sinh lactic acid cua cac chiing nay Nguyen Thi Hoai Ha va ddng tac gia (2002) da nghien ciru kha nang sinh tdng hgp bacteriocin ciia loai Lb plcinlanim L24 Nguyen Vii Tudng Vy va ddng tac gia (2007) da khao sat kha nang chiu mudi mat. acid \a khang
DdThj Bich Thiiy ei at.
khang sinh cua mdt sd vi smh vat diing lam nguyen lieu san xuat probiotic dudng udng. trong do cd Lb acidophilus, Lb casei, Streptococus faecaiis. Ho Van Thao va Hoang Qudc Khanh (2006) ciing da phan lap va tuyen chon mot sd ddng Enterococcus cd kha nang chm dugc cac dieu kien dudng rugt, kha nang ket bam, kha nang sinh bacteriocin de lam che pham probiotic
De gdp phan cung cap them cac thdng tin ve tiem nang img dung cua he vi khuan lactic, chiing tdi lien hanh dinh danh va nghien ciru kha nang len men lactic va mdt sd tiem nang probiotic ciia chung DC2 phan lap duac tir san pham dira cai.,
V A T L I E U VA PHUONG PHAP NGHIEN CUH
\ ' a t lieu nghien eiiu
Chiing DC2 dugc phan lap iir san pham dua cai chua tren dia ban thanh phd Hue
Phuo'ng p h a p nghien CUTI
Phmmg phdp xtic dinh ham lugng lactic acid trong dich nuoi cay hang sac ky long cao ap (HPLC)
Ham luong lachc acid Irong dich ihu dugc xac dmh bang sac ky long cao ap (HPLC) tren cdt C18 - Thermo, budc sdng 220nm. tdc dd ddng 1 ml/phiil Phuong phdp vi sinh
Phun logi sg hg chung DC2 den cap itg chi bang each xdc dinh mgt so dgc diem sinh ly, sinh hoa vd dimg phuotigphdp logi trir
Tien hanh cac khao sat su phal tnen ciia chung \i khuan DC2 tren mdi trudng MRS vdi cac dieu kien Ihay ddi gdm (Cho va Do, 2006, A.\elsson. 2004) Khcio sdl kha ndng phdl men Irong cdc dieu kien mdl truang khac nhau
Cay mau tang sinh vao mdi trudng long can khao sat tuong iing (pH d 4,4 va 9,6; nhiet dd 10"C
\ a 4.>"C. ndng do mudi NaCI 6,5% va 18% ) vdi ti le 5%, mau dugc do mat dd quang (OD) d 0 gid va sau 24 h a budc sdng 600nm; neng cac khao sat a nhiet dd 1 ()"C duac do sau 96 h.
Klicio .sdl klia niing smh CO: lie ten men glucose Tien hanh nudi cay vi khuan trong dng nghiem chira mdi trudng MRS long, pH 6.2 Cho dng Durham vd trimg iip ngugc vao, u a 37"C trong 48 h va quan s;il Sir ndi len ctia dng Durham se cho ket qua duong linh
!46
Phdn logi chiing DC2 bdng pfiurmg phdp sinh bgc phdn tir
Tdch ADX
Cho dich huyen phti vi khuan vao tube Eppendorf 1.5 ml da cd san 200 pi dung djch Instagene^^' MaxTrix, vortex 10 giay va sau dd spin nhe Hdn hpp mau tiep tuc dugc ii d 56"C trong 30 phiit Sau do, vortex mau trong 10 giay va dun sdi mau d I00"C trong 8 phut, rdi li trong nudc da trong 5 phiit. Sau khi Iy tam 13.200 vdng/phiit trong 5 phiiL pha loang 10 Ian dich ndi thu dugc de ihue hien phan img PCR.
PCR vd giai Irinh lu ADN 16S
Cho 5 pi dich ADN ctia vi khuan vaa 20 pi hdn hgp PCR(''^16S-PCR) (Cdng ty Nam Khoa, Thanh Phd Hd Chi Mmh) va thuc hien phan iing PCR, theo chuong trinh luan nhiet nhu sau, I chu ky 9 5 T trong 10 phiit; 40 chu ky gdm cac giai doan' 94°C trong 30 giay, 60"C trong 30 giay, 72''C trong 1 phiil va 1 chu ky 72"C trong 10 phiit San pham PCR duac kiem tra bang phuang phap dien di tren gel agarose 2%!. San pham PCR cd do dai khoang 500 bp duoc tmh sach va tien hanh giai trinh tu tren he thdng ABI 3I30XL
Phuo'ng phdp khdo sdl khd ndng chiu acid Rua sinh khdi te bao bang dem phosphale 0,1M pH 7 va tai huyen phii trong dem nay. Iron Iml djch huyen phii vdi 24,5 ml dung dich NaCI 0,2 % cdpH 2 va phan phdi vao cac dng eppendoff (Iml /ong).
Xac dinh sd lutmg te bao sdng trong cac ong eppendoff a cac thai gian khac nhau (0 h, Ih, 2 h va 3 h) bang phuong phap Kock.
Khdo sdt khd ndng chiu muoi mat (Gilliand va Walker, 1990)
Nudi cay vi khuan trong mdi trudng MRS long cd bd sung mudi mat 0,3% (No 3, Merck) d 37"C trong 4 h. Mat dd te bao trong mau duac xac dinh bang each xac dmh mat do quang d budc sdng dOOnm, Mirc dd chiu mudi mat dugc xac dinh bang thdi gian de tang ODhoi)i.m sau khi nudi 4 h so vdi ban dau Ien 0,3 dan vi
Khdo sdl khd nang ket dinh ctia vi khuan lactic Khao sdl khd ndng lu kel dinh
Sinh khdi te bao cua vi khuan thu duac rira 2 lan bang dem PBS (8 g NaCI, 0,2 g KCI, 1.44 g Na.HPO*, 0,24 g KH^PO*, bd sung nudc cit du I lii) pH 7.2 vd triing. sau dd, dugc tai huyin phii trong
Tap chi Cong nghe Sinh hoc U(l) 145-152. 2013
d?m PBS (ODsoonm =1) ( O p ban diu). Giir huydn phii nay d 3 7 X trong 5 h de tao dieu kien cho cac vi khuan lactic ttr ket dinh va lang xudng va do OD dich tren bS mat, Kha nang tu ket dinh la phan tram dg giam ODnonnm dich be mat ciia mau da giir 5 h so vdi ban dau,
Khdo sdl kha ndng dong ket dinh v&i S aureus
Sau khi nia 2 Ian bang dem PBS. sinh khdi tdbao cua chung DC2 va S. aureus duoc tai huyen phii den ODenonni^l- Ddng thdi trdn Ian hai huyen phii nay vdi the tich bang nhau, Tien hanh do ODisoonm dung dich tren hk mat sau khi giir huyen phii ciia moi chung \ a hdn hgp huyen phii ciia hai chiing d 37"C rrong 5 h, Kha nang ddng ket dinh dugc tinh Iheo cdng thirc.
(A^ + A , . ) / 2 - A , , , Ty leddng ket dinh (%) = (-
(A;,, + A J / 2
au 5 hcLia vi khuan lactic. Ay is ODeoonm sau 5 h ciia S ac a A X— la ODsaonm sau 5 h ciia v
Khao sdl khd ndng kel dinh vdi dung moi Rira sinh khdi te baa chung DC2 bang dung dich KNOi O.IM, pH 6,2 va tai huyen phii vao dung dich nay den ODboonm ^ 1- Cha Iml mdi Ioai dung mdi (xylene, chloroform, ethyl acetate) lan lugt vao 3 dng nghiem chira 3 ml huyen phii te bao, vortex
trong 2 phuL sau do de yen 20 phiil o nhiet dd phdng, tach pha nudc va do dp hap thu Kha nang ket dinh vdi dung mdi dugc danh gia la phan tram dp giam ODboonm ciia pha nudc thu dugc trong cac dng nghiem cd dung mdi sau 20 phiit giir so vdi ban dau (OD.o()„.= I)
Bang 1. K^tqua dmh danh so bo cac chung vi khuan DC2
Kha nang Phat trien 6" Phat trien CT Phat tnen a Phat trien 6 sinh COi lO-C 45°C pH 4.4 pH 9,6
KET QUA VA THAO LUAN Phan loai str bg chiing DC2
Kha nang phat tnen cua chiing DC2 o cac dieu kien pH, nhiet dp, ndng do mudi khac nhau va smh CO2 dugc the hien tren bang !.
So sanh kdt qua tren bang I vdi bang phiin loai VI khuan d i n ehi cua Axelsson (201)4), chdng idi nhan thay rang cd hai dac diem the hien su khac nhau ro ret ve dac tinh sinh ly, sinh hda cua chung dugc khao sat so vdi chi Carnohaclenuin
!a DC2 cd th6 phat tridn d 45°C va khdng phat trien d pH 9,6 Vi vay, chung DC2 thudc chi Lactobacillus,
Phan loai chiing DC2 biing giai trinh tir gene I6S rDNA
Ket qua dugc tra ciru tren ngan hang gen ctia NCBI Ihdng qua chuang trinh BLAST va dua vao sy tuong ddng trong trinh tu cua doan gen de dinh danh din loai (Hinh I) Kdl qua cho thay, trinh tu
DNA 16S cua chung DC2 khi ddi chieu vdi trinh tu gen ctia chung Lb ptanlarum NWL61 o ngan hang gen cd dd ddng nhat dat den 99%) Vi vay, chung DC2 thudc loai Lb ptanlarum.
Xac dinh kha nang sinh lactic acid trong djch nudi ea\ cua chiing Lb. plantaruin DC2
Ham luong lactic acid trong mdi truang sau 72 h nudi cay Lb. plantarum DC2 xac dmh duoc bang HPLC (Hinh 2) la 23.41 g/'l (>20 g/l). So sanh kel qua nay vdi mdl sd cdng bd trudc day cho ihay chung nay len men sinh lactic acid luong ddi cao Chang ban nhu. chiing L lactic subs laclis cd kha nang sinh 13.7 g/l lactic acid trong mdi truong nudi ciy (Cock, Rodriguez de Stouvenel, 2006). Kel qua nghien ciru cua Nguyen The Trang va Trim Dinh Min (2008) fren mdi trudng MRS cd bd sung 15 g/l glucose, pH 6.0. sau 48 - 60 h nudi c;i\. (T 3 0 " C chung VI khuan lactic HNl leo ihe san sinh I0,'J4 g l Vdi chung l l \ 3 4 o ciing mdi trucmg \a thoi gian nudi c i \ nhung nhict dd nudi 37"C hrcmg lactic acid dat 12.76 g I- ^
Do Thi Bich Thiiy el al.
GCTGGCGGCGTGCCTAATAC.\TGCA,AGTCGAACO,A.ACTCTCaT,ATTCATTGOTCCTTGCA irATOT.ATTTAr ATTTG AGTOAOTGGCG A AGTCGTC AGT A AG ACGTOOC AAACCTGCCCA GAAGCGGCGCATAAC\CCTGGV\ACAG.s,TGCTAATACCCCATAACAACTTGG.ACCGCAT OGTCCGAGTTTGAAAGATGCCTTCGGCTATCACTTTTGGATGGTCCCGCGOCGTATT.AGC TAGATGGTGGOGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGACACGGT,A,AT CGGCCACATTGGGACTGAG.ACACGGCCCAAACTCCTACGCGAGCCACCAGTAGGGAATC TTCCACAATCGACOAAAGTCTGATGOAGCAACGCCGCGTGAGTGA.AGAAGGGTTTCGGC TCGTAAAACTCTCTTGTTAAAGAAGAACATATCTGACAGTAACTaTTCAGOTATTCACCC TATTTAACCAGAAAGCCACGCCTAACTACGTOCCAGCAGCCGCGGTA
Hinh 1. Trinh tir mot doan DNA 16S cua DC2
il" -M
Hinh 2 Sac ki do xac dmh ham lucng lactic acid, A Mlu chuan, B. Dich len men ben Lb ptanlarum DC2
Tgp chi Cong nghe Sinh hpcll{\) 145-152, 2013
Mot so tinh chat the hien tiem nang probiotic cua Lb. plantarum DC2
Kha ndng chiu muoi mdl ctia Lb. ptanlarum DC2 Ket qua khao sat kha nang khang mudi mat ciia Lb. plantarum DC2 thdng qua kha nang phat InSn ciia nd a ndng do mudi mat bd 0.3% tuang ddi cao;
sau 4 h AOD do d budc sdng dOOnm dat 0,377. Cac tac gia Liong va Shah (2005) da ti&n hanh khao sat kha nang tang trudng ciia vi khuan lactic trong mdi tnrdng MRS ed 0,3% djch mat bd, ket qua cho thiy tdc do tang OD d budc sdng 600nm trong khoang 0,068 (Lb. casei ASCC 290) ddn 0,127 (Lb. casei ASCC 1520) dan v|/gid, tuong duong vdi AOD khoang 0,275 - 0,508 sau 4 h. Mota va ddng tac gia (2006) Cling da tien hanh nghien ciiu kha nang khang mudi mat ciia 3 chung vi khuan Lb acidophilus. Lb acidophilus 2MI4E bi iirc che manh, sau 4 h trong pha log chi tang dugc khoang 0,2 don vi OD 600 d ndng dd mat bd 0,3%, Lb acidophilus 2G14E bi iic che trung binh thi tang dugc khoang 0,4 va Lb.
acidophilus 5C14E it bi irc che thi tang dugc khoang 0,5 Tir do cho thay rang chiing Lb plantarum DC2 cd kha nang chiu mudi mat tdt.
Kha nang chiu acid ctia Lb. plantarum DC2 Xac dinh sd te bao sdng ciia Lb. ptanlarum DC2 qua cac mdc thdi gian lien tuc tir 0 gid den 3 gid trong mdi trudng pH 2 de khao sat kha nang chm acid (Hinh 3), Sau 1 h, 2 h va 3 h u a pH 2 lugng ia bao sdng cua chung nay van cdn tuong img la 7.517 CFU/ml (85,56 % sa vdi ban diu). 6,557 CFU/ml (74,63% so ydi ban diu) va 6,279 CFU/ml (71,47 % so vdi ban dau). Ket qua nay cha Ihay Lb. plcmlarum DC2 cd kha nang chiu acid cao so vdi mdt sd cdng trinh da cdng bd. Lb acidophilus ATCC 4962, Lb casei ASCC 290 va Lb.ca.sei ASCC 292 sau 2 h nudi cay trong pH2 giam tir 10 CFU/ml xudng 7 CFU ml (70% so vdi ban diu) (Liong, Shah, 2005), Kha nang chiu acid cua mdt sd chung Lactobacillus cdng dugc nghien ciru bdi Maragkoudakis va ddng tac gia (2006). Ket qua cho thay cac chung cd kha nang khang acid manh nhat la Lb. paracasei subsp paracasei ACA-DC 130. Lb planianiin AC.A-DC 146, Lb. rhamnosiis ACA-DC 112. vdi mirc giam CFU/ml lir 8.6 xudng cdn lan luot la 6,8: 5.7 va 7.1 sau 3 h u o pH2. ngoai ra, mdt sd chung trong nghien ciru ciia nhdm tac gia nay khdng cd kha nang sdng sdt d pH2 sau 1 h
10 8,786
& 2
6,279
I
Tho'i gian nuoi cay (gio) Hinh 3 Kha n^ng sdng sot cua L ptantanim DC2 d pH 2 theo ihoi gian
Khd ndng hdm dinh cua Lb. plantarum DC2 Kel qua khao sat vc kha nang bam dinh cu;
ptantanim DC2 rat kha quan (Bang 2) Orlowsk va Beelccka (2006) nghien ciru nang lu kel dinh cua Lactobacillus Bifidobacienum da nhan thay cd su bien ddnij
trong kha nang tu ket dinh cua cac chung, 5 chLtng dugc khao sat trong thi nghiem cd kei kiua nr kci dinh la $.^"'>>. I5";..''23%. 75% va 77",, ,, nhiel do phdng Ket qua nghicn cuu kha nang ur kci dinh cua cac chuny tiitidobaetenum o -" I hoi Rahman va ddng lac gia (200S} nam troii;j khoang (1.9% - I3.2".., Maria (200(.| du.i i .t ki-i qua \c lu
bam dinh ciia I 1 chiing Lactobacillus gdm cac loai Lb. casei; Lb Johnsonii; Lb. paracasei; Lb.
rhamnosus va Lb. plantanum vdi ket qua chu yeu nam trong khoang tir 20%) den 40%o; Khi tdng hgp nhieu dac diem de de xuat chimg cd tiem nang lam probiotic, tac gia nay da de xual 4 chung gdm Lb Bang 2, Kha nang bam dinh ciia Lb plantarum DC2
Do Thj Bich Thiiy el al.
ptanlarum BFE 5759, BFE 1684, BFE 1685 va L.
johnsonii BFE 6128, cac chung nay cd ty le tu ket dinh lin lugt la 4 0 % , 2 0 % , 2 8 % , 3 5 % So sanh vdi cac ket qua da cdng bd cha thay Lb. ptanlarum DC2 cd kha nang tu kdt dinh kha cao (38,57%) nen cd the su dung de Iam probiotic.
Kha nang bam dinh (%) 38,57
24,23 61,46 43,27 74,81 Tieu c h i danh gia
Kha nang tu ket dinh
Kha nang d6ng ket dinh voi S aureus Kha nang bam dinh voi dung moi xylen Kha nang bam dinh voi dung moi ethyl acetate Kha nang bam dmh vffi dung moi chloroform
So vdi mdt sd cdng trinh da cdng bd, kha nang ddng ket dinh vdi vi khuan gay benh cua L.
plantarum DC2 (24,23%) cd the lam tieu chi danh gia tiem nang probiotic Collado va ddng tac gia (2007) da khaa sat kha nang ddng ket dinh cua cac chiing vi khuan lactic cho thay sau 4 gid, ty Ie ddng ket dinh trong khoang 6% den 27%; Kos va ddng tac gia (2003) da nghien ciiu kha nang ddng ket dinh cua Lb acidophilus M92 (chung nay dang duoc su dung san xuat ehe pham probiotic) vdi Enterococcus faeciiim L3, E.scherichta coh 3014, Salmonella cdng bd kdt qua tir 15,11 % ddn 19.46%,
Kha nang bam dinh cac dung mdi xylen, ethyl acetate va chloroform cua L. fermentum DC2 lan lugt la 61,46%; 43,27% va 74,8% (Bang 2) Rahman va ddng tac gia (2008) da cdng bd kha nang ky nudc cua mdt sd vi khuan Bifidobacterium bien dgng rit ldn (14,4% - 97,3%). Pelletier va ddng tac gia (1997) cho ket qua khao sat su bam dinh vdi hexadecane (dung mdi khdng phan cue) ciia mdt sd chung L casei subsp casei, L paracasei subsp. paracasei, L rhamnosus la 5,8% -26,5%i. Theo Maria (2006), kha nang bam dinh ethyl acetate cua mdt sd chiing L johnsonu, L plantarum, L paracasei, L. casei nam trong khoang 0 - 79,2%j. Kha nang bam dinh vdi dung mdi ethyacetate cua Lb casei cho k6t qua n i m trong khoang 10 - 60% (Provencio el al, 2009). Vd Nhan Hau va ddng tac gia (2005), cho thiy kha nang bam dinh chloroform cua mdt sd chung vi khuin lactic dao ddng tir 0,51% ddn 87,32%. Tir cac kdt qua da cdng bd cha thay chiing L plantarum DC2 cd kha nang bam dinh dung mdi rat cao nen cd tidm nang probiotic Idn.
KET LUAN
Chung DC2 dugc phan lap hi dua cai thudc Ioai Lactobacillus plantanum. Chiing nay cd kha nang len men sinh lactic aciti cao Ndng do lactic acid trong moi trudng dat 23,41 g/l sau 72 gid nudi cay trong MRS long cd bd sung glucose (20 g/i) d 37''C, Ben canh do, chung DC2 cdn cd kha nang sdng sdt trang mdi trudng pH2 (sd te baa sdng sdt so sau khi u d pH2 trong mdt gid, hai gid va ba gid Ian lugt la 85,56 % , 74,63% va 71,47 % so vdi ban diu), cd kha nang chiu mudi mat (AOD 600 dat 0,377sau 4 gid trong mdl trudng cd ndng dg mudi mat 0,3%) va kha nang bam dinh tdt (kha nang tu ket dinh va ddng ket dinh vdi Staphylococcus aureus la 38,57 % va 24,23 %, kha nang bam dinh vdi cac dung mdi xylene, ethyl acetate va chloroform Ian lugt !a 61,46
%, 43,27 % va 74,81 %. Tren ca sd cac dac diem da xac dinh, chiing L plantarum DC2 cd the dugc nghien cihi ung dung de san xuat che pham probiotic,
TAI LIEU THAM KHAO
Nguyen Thi Hoai Ha, Pham Van Ty, Nguyen Thi Kim Quy (2002) Nghien ciru kha nang sinh long hop bacterioxin ciia Ioai Lactobacillus plantarum L24, Tap chi Di truyen hgc vo img dung. Chuyen san Cong nghe sinh hoc 2002, Ha Ngi, tr 47-52
V5 Nhan Hau, Ly Mai Hucmg (2005) Nghien cuu tinh chat hda Iy thanh te bao vi khuan lactic va anh hudng ciia chiing den ihanh phan he nhii tucmg. Tap chi Khoa hoc ky thudt nong nghiep, 4.327-332,
Tap chi Cong nghe Smh hoc 11(1); 145-152. 2013
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S T U D Y ON P R O P E R T I E S O f LACTOBACILLUS PLA.STARUM D C 2 I S O L A T E D FROM T R A D I T I O N A L L A C T I C F E R M E N T E D P R O D U C T " D U A C A I " IN H U E CITY, V I E T N A M
Do Thi Bich T h u y ' % Phan Thi Be', Tran Thi Ai Luyen' ' Hue Universtly of Agriciillure and Foresiiy ' Hue Tourism College
SLMMARY
A laclic bacterium strain DC2 isolaled from "Dua cai". a traditional lactic fermented produci in Hue eily, Vietnam, was firstly identified by several biochemical and physiological characterizations (investigation of Ihe growth m pH 4 4 and 9 6, al temperature 10"C and 45"C, 6.5% and 18% of NaCI concentrations, ability to produce COi) The obtained results revealed Ihat it belongs to genera Laciobacittiis genus Phylogenetic analysis showed Ihal llns strain was closest to Lb plantarum named Laciobacittiis ptanlarum DC2 The lactic acid conieni from strain DC2 was 23 41 g/I after growing in MRS medium for 72 hours at 37"C Some probiotic potennal properties of strain DC2 were investigated. The number of survival cell after incubation at pH2 for one hour, two hours and 3 hours are 85.56%, 74 63% and 71 47%, respectively In addition, Ihe bile sah tolerance of this strain was high After 4 hours of incubation in 0 3% of bile sah soluUon AOD in 600 nm was 0,377 h exhibited the ability of autoaggregation to be 38.57% and Ihat of coaggregation with Staphylococcus aniens to be 24 23%. Its abilities of adhesions to xylene, ethyl acetate and chloroform were 61 46%,4.V27%and74 81 ';;>, respectively
Keyword.s: Identifieaiion, isotcilum. taciu acidfei mention. Laciohacillus fermeniuni. probiotic
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