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EXOPOLYSACCHARIDES
LI NI U
Ngy nay, vi s tin b ca khoa hc cng ngh, con ngi ngy cng tip cn gn hn, hiu bit nhiu hn n th gii sinh vt ni chung v h vi sinh vt ni ring. V, vi nhng bc tin ca mnh, cc nh khoa hc nghin cu thnh cng rt nhiu nhng sn phm c sn xut t vi sinh vt, gp phn quan trng trong nn cng nghip sn xut, trong y hc, trong x l mi trng,....V mt trong nhng sn phm quan trng l exopolysaccharides.
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HNH 1: Biu ng ng mc ca EPS sn xut bi L. subsp.bulgaricus delbrueckii RR 30 g ca Bacto-casitone mi lt nh mt hm s ca nhit v pH. ng ng mc vi nhng con s ng k khc nhau (P <0,05). Cc ng cho l ranh gii thit k, tc l, iu kin th nghim bn di dng ny khng c trong thit k. Nng sut thc t theo nhng iu kin ny l 354mg EPS l, iu ny chnh xc 95% so vi d an ( l thuyt).Mt l ca cc kt qu ch ra rng c th thu c sn lng EPS cao hn nu nng Bacto-casitone ln hn 30g l (hnh 2). kho st, mt th nghim c tin hnh nhit ti u (38C) v pH (5.0) vi nng Bacto-casitone 40g/l.Sn xut EPS di nhng iu kin ny c tm thy l 324g/l, thp hn sn lng ca EPS c sn xut trong iu kin ti u (Bng 1).Khi kt qu ny c a trong phn tch, d an ti u v ngha mi quan h gia cc iu kin tng trng v phn ng bin i (bin d) khng c s thay i. Bng 2 cung cp cc h s ca cc m hnh thng k thu c t nhng phn ng b mt, cng nh ngha thng k ca mi k.
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a. mi th nghim vi s ln khc nhau cho thy mt tp hp cc iu kin th nghim b. chy biu th th t m trong cc th nghim c tin hnh
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HNH 2: th ba chiu ca EPS sn xut bi L. delbrueckii subsp. bulgaricus RR pH 5.0 l mt hm s ca nhit v nng Bacto casitone.
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BNG 2: H s xc nh m hnh phn ng a Y = 0 + 1(T 40) + 2(pH 5) + 3(casitone 20) + 4(T 40) (pH 5) + 5(T 40)(casitone 20) + 6(pH 5)(casitone 20) + 7(T 40)2 + 8(pH 5)2 + 9(casitone 20)2, trong T l nhit ln men ( C), pH l pH ca mi trng trong qu trnh ln men, v casitone l s gam Bacto-casitone mi lt trong mi trng pht trin. b. Thi gian , 90% ca glucose trong mi trng pht trin c s dng bi cc sinh vt. Thi gian thit b u cui c mt thiu ht ng k ph hp (P <0,05) vi m hnh b mt phn ng. c. Khc bit ng k mc P <0,05 d. Khc bit ng k mc P <0,001 e. NI, khng c bao gm. Bao gm cc h s ny trong m hnh khng ci thin ph hp f. Khc bit ng k mc P <0,01. g. H s hi quy ca m hnh d on bin phn ng c th. h. gi tr P cho cc m hnh d on bin phn ng c th. Nhit ti u sn xut EPS nm trong khong nhit tng trng ti u ca RR (37-450C). Th nghim cng b tc ng ca nhit trong sn xut EPS bi vi khun acid lactic b tri ngc li.Mt vi bi bo co tm thy sn lng EPS ln
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HNH 3: ng vin l sn xut (sn phm) vi 30 g ca Bacto-casitone mi lt nh mt hm s ca nhit v pH. ng ng mc vi nhng con s ng k khc nhau (P <0,05). Cc ng cho l ranh gii thit k, tc l, iu kin th nghim bn di dng ny khng c trong thit k. Mt s t th nghim c lm nghin cu tc ng ca pH trong sn xut EPS bi actobacilli, v khng c kim sot bi sinh vt s dng trong nghin cu. Mozzi o lng lng polysaccharide c tng hp ti a ( 488mg l) v s lng t bo cao nht ti im pH = 6.0 ca vi khun . casei CR 87. Thm mt th nghim khc , s lng EPS (mg/l) iu kin pH c kim sot cao gp 3.6 ln so vi iu kin pH khng c kim sot. Trong nghin cu kim sot pH vi L.sake 0-1, pH ti u sn xut EPS l 5.8, nhng s lng t bo cao l pH = 6.2. T kt qu , cc nh nghin cu kt lun rng s bin i ng to EPS t hiu qu hn ti pH = 5.8, nhng lng ng to sinh khi cao l pH = 6.2.
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HNH 4: Thng tin ca EPS sn xut, trng lng kh t bo, v glucose s dng bi L. delbrueckiisubsp. bulgaricusRR pht trin trong iu kin ti u cho sn xut EPS (38 C, pH 5,0, 30-g/liter Bacto-casitone). , EPS; , glucose, , trng lng t bo kh. i vi 1 s vi khun sn xut EPS nh Xanthomonas, Pseudomonas, Rhizobium spp, s gii hn nitrogene l kt qu trong qu trnh tng sn xut EPS nhng n khng c p dng dng RR. Sn xut EPS ti u xy ra khi mi trng cha gp 3 ln nng Bacto-casitone (30g l) p ng nhu cu sinh trng (10g/l). S tng lng Bacto-casitone trong SDS n 40g/l khng c kt qu trong s tng mc sn xut EPS.iu c th din ra c bi v nhng cht dinh dng khc c nh ra gii hn va trong m nui cy. H phng trnh bc 2 dng trong ci biu th hin s tc ng ln b mt ca th nghim c s dng trong ci nghin cu ny. oi phng trnh ny rt l logic bi v nhng tha s tc ng n s sinh trng ca vi khun thng cho thy c 1 im ti a. S phn tch thng k cung cp thng tin v s nh hng ca iu kin sinh trng da trn nhng thay i ca tc ng. Mi quan h ng k c tm thy gia nhit v sn lng EPS. Ni chung, mi quan h gia nhit v s sinh trng ca vi khun cho thy c1 im ti a, nhng khng i xng, cng nh mi quan h c kho st y. Mi quan h gia pH v s sinh trng ca vi khun cho thy l mt vng tr s cc i. Trong nghin cu ny, c v ph hp v pH khng c tc ng ng k n sn xut EPS bi v khong s dng nghin cu (4 n 6) c th hin l ng tng i ph ng trn ng cong ca th. Trn thc t, hnh 1 th hin rng nhit ti u v iu kin pH ca qu trnh sn xut EPS tn ti trong ci ng elip hnh thnh bi 36-39oC v pH t 4.5 n 5.5. Cui cng,
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During recovery
97.00
367.84c
35.68
100.0
5.32
6,647.27c
35.36
99.1
After recovery
Freeze-dried precipitate
964.0022.0e 29.20f
81.84
a. Nng sut (grams) = EPS (gram mi lt) khi lng trung bnh (lt) b. Thu hi (%) = g EPS / gram EPS c trong canh trng ln men 100. c. Trong miligam trn mt lt. d. Sau khi siu lc v diafiltration e. Trong mg mi gram. f. EPS c mt hm lng protein 1,42% (0,43 0,11 g) v m 0,3% (0,0073 0,0005 g). Nng sut, nh ngha l s lng sn xut ti a trong 1 khong thi gian ti thiu, l bin s quan trng t cng ngh trin vng. xc nh iu kin sn xut ti u, cc s tc ng thay i nng sut c kt hp li, dng cho d kin nng sut, c tnh ton bng tng trng lng ca nng EPS di nhng tc ng ca mt iu kin cng vi thi im kt thc. Bng cch s dng chng trnh o s ti u ha
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TI LIU THAM KHO Bradford M. M.(1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248254. Breedveld M. W., Zevenhuizen L. P. T. M., Canter Cremers H. C. J., Zehnder A. J. B.(1993) Influence of growth conditions on production of capsular and extracellular polysaccharides by Rhizobium leguminosarum.Antonie Leeuwenhoek Cerning J. (1990) Exocellular polysaccharides produced by lactic acid bacteria. FEMS Microbiol. Cerning J., Bouillanne C., Desmazeaud M. J. (1988) Extracellular polysaccharide production by Streptococcus thermophilus. Biotechnol. Lett.10:255260. Cerning J., Bouillanne C., Landon M., Desmazeaud M. (1992)Isolation and characterization of exopolysaccharides from slime-forming mesophilic lactic acid bacteria. J. Dairy Sci. 75:692699. Cerning J., Renard C. M. G. C., Thibault J. F., Bouillanne C.,Landon M., Desmazeaud M., Topisirovic L. (1994) Carbo n source requirements for exopolysaccharide production by Lactobacillus casei CG11 and partial structure analysis of the polymer. Appl. Environ. Microbiol.60:39143919. Dubois M., Gilles K. A., Hamilton J. K., Rebers P. A., Smith F.(1956) Colorimetric method for determination of sugars and related substances. Anal. Chem. 28:350356. Gancel F., Novel G. (1994) Exopolysaccharide production byStreptococcus salivarius ssp. thermophilus cultures. 1. Conditions of production. J. Dairy Sci. 77:685688. Garcia-Garibay M., Marshall V. M. E. (1991) Polymer production byLactobacillus delbrueckii ssp. bulgaricus. J. Appl. Bacteriol. 70:325328. Grobben G. J., Sikkema J., Smith M. R., de Bont J. A. M. (1995)Production of extracellular polysaccharides by Lactobacillus delbrueckii ssp.bulgaricus NCFB 2772 grown in a chemically defined medium. J. Appl. Bacteriol. 79:103107. Gruter M., Leeflang B. R., Kuiper J., Kamerling J. P.,Vliegenthart F. G. (1993) Structural characterisation of the exopolysaccharide produced by Lactobacillus delbrueckii subspeciesbulgaricus rr grown in skimmed milk. Carbohydr. Res. 239:209226.
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