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1. doi: 10,1128 / AEM.71.12.8383 8389,2005 Appl. Mi

trng. Microbiol. - Thng 12 nm 2005 vol. 71 khng c. 12 8.3838389

Trong Vitro Xc nh tnh Prebiotic oligosaccharides ngun gc t sn xut Orange Juice By-Sn phm Sui
1. 2. 3. 4. 5. 6.

K. Manderson 1 , M. Pinart 1 , KM Tuohy 1 , Chng ti Grace 1 , AT Hotchkiss 2 , W. Widmer 3 ,

7. MP Yadhav 2 , 8. GR Gibson 1 v 9. RA Rastall 1 , *

+ Tc gi chi nhanh Trng Khoa hc Sinh hc Thc phm, i hc Reading, PO Box 226, Whiteknights, c RG6 6AP, Vng Quc Anh 2. 2 ng khu vc Trung tm Nghin cu, Dch v Nghin cu Nng nghip, B Nng nghip M, Wyndmoor, Pennsylvania 19038 3. 3 Citrus v Phng th nghim Subtropical Sn phm, dch v nghin cu nng nghip, B Nng nghip M, 600 Avenue S, NW, Winter Haven, Florida 33881
1.
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Mc tip theo

TM TT
Ti sn ln men ca oligosaccharides c ngun gc t pectin v cam c nh gi trong nn vn ha pha trn ca vi khun phn. V cam phn oligosaccharide c glucose trong rhamnogalacturonan v oligosaccharides pectic xylogalacturonan. Hai mi bn ting ng h, nhit v pH c kim sot, khng c khuy k kh phn l nn vn ha c s dng xc nh hiu qu m cc oligosaccharides bt ngun t cc sn phm cam c trn cc thnh phn ca h vi sinh vt phn. Cc hiu ng c nh gi thng qua lai ti ch hunh quang xc nh nhng thay i trong cc qun th vi khun, ln men sn phm cui cng c phn tch bng sc k lng hiu nng cao nh gi nng acid bo chui ngn, v sau , mt ch s prebiotic (PI) c xc nh. Oligosaccharides pectic (POS) c th tng cc s bifidobacterial v Eubacterium rectale , mc d kt qu l mt ch s tin sinh thp hn t qu trnh trao i cht fructo-oligosaccharide.Orange sut phn chiu vn duy tr s tng trng ca hu ht cc qun th vi khun v a ra mt PI tng t nh tinh bt ha tan. Qu trnh ln men ca POS dn n s gia tng s rectale Eubacterium v ng thi tng sn xut butyrate. Trong kt lun, nghin cu ny ch ra rng POS c th c mt tc dng c li trn h vi phn, tuy nhin, mt hiu ng c in tin sinh khng c tm thy. S gia tng dn s Eubacterium rectale c tm thy, v mc butyrate tng ln, l li ch tim nng cho cc my ch. Khi nim v sc khe ng rut iu ch thng qua ch n ung khng phi l mi v ngy tr li vo u ca th k 20. Tuy nhin, n ch l gn y

rng l do c bn m thanh khoa hc c xut v iu tra. Ba cng c iu ch vi xut hin ( 40 ): vic b sung cc vi sinh vt sng ngoi sinh thc phm (tc l, cc ch phm sinh hc), kch thch chn lc ca s tng trng v hot ng ca vi sinh vt bn a mang li li ch cho ng rut (tc l, prebiotics), v s kt hp ca c hai phng php tip cn (tc l, synbiotics). Tt c ba phng php tip cn c gng tng s lng vi khun c xem l tch cc cho sc khe tiu ha ca con ngi, thng l lactobacilli v bifidobacteria. Prebiotics c nh ngha l "thnh phn thc phm khng tiu ha c chn lc kch thch mt s hn ch ca vi khun i trng, ci thin sc khe ch nh" ( 12 ). Bifidobacteria v lactobacilli c gi trc tip c ch s tng trng ca vi khun gy bnh, chng hn nh mt s loi clostridia (v d, Clostridium difficile , C. perfringens ) v gy bnh Enterobacteriaceae , thng qua vic sn xut cc axit bo chui ngn, lm gim pH i trng, sn xut cc hp cht khng khun, v / hoc cnh tranh cho nn tng trng v cc trang web kt dnh ( 13 , 23 , 27 ). Tnh cht bifidogenic ca mt lot cc prebiotics c xc nhn trong c th trong cc nghin cu thc n ca con ngi ( 11 , 37 ). Nhng prebiotics bao gm inulin, fructo-oligosaccharides (FOS), galacto-oligosaccharides, v lactulose ( 42 , 43 , 25 , 44 ). Mc ch ca cng vic ny l nh gi cc tnh cht tin sinh hc cc oligosaccharides chit xut t v cc sn phm sn xut nc cam. Citrus d lng cn li sau khi khai thc nc tri cy i din cho khong mt na khi lng tri cy m t, v khong mt na d lng ny l v ( 2 ). Hn 2.000.000 tn bt vin bt cy c mi c sn xut hng nm cho thc n gia sc t d lng ch bin cy c mi, m l mt sn phm gi tr thp. Pectin, mt sn phm gi tr cao hn, c sn xut t v cam qut, nhng nhu cu th trng cho pectin l thp so vi pectin tim nng c th c sn xut t cc ngun cung cp trn ton th gii ca d lng ch bin cy c mi. Oligosaccharides pectic (POS) c sn xut t pectin thng mi trong mt l phn ng mng t bo enzyme ( 30 ) v sau nh gi tnh cht tin sinh ( 29 ).Cc oligosaccharides pectic ny c mt tim nng thp tin sinh so vi FOS, mc d h c chn lc ln men hn l cc pectin cha m ( 29 ). Oligosaccharides pectic colonocytes cng c bo v chng li verocytotoxins Escherichia coli ( 32 ) v kch thch apoptosis trong t bo ung th tuyn ngi i trng ( 31 ). y, chng ti nhm mc ch tm thy mt cch chi ph-hiu qu hn sn xut oligosaccharides prebiotic pectic trc tip t v cam sut phn chiu (OA).Fructo-oligosaccharides, cc prebiotics rng ri nht nghin cu v cc

nh lnh o hin ti th trng, c chn nh mt iu khin tch cc cho nghin cu qu trnh ln men phn trong ng nghim, trong khi tinh bt, mt carbohydrate nonprebiotic, c s dng nh mt iu khin khng chuyn. trc Mc sau

TI LIU V PHNG PHP


xc nh tnh chn lc ca qu trnh ln men ca vt liu kim tra so vi FOS v tinh bt, kim sot pH, khuy ng nn vn ha hng lot tim slurries con ngi phn c s dng. Vi khun c thc hin bng cch s dng hunh quang lai ti ch vi cc u d oligonucleotide nhm mc tiu c th 16S rRNA khu vc ca cc vi sinh vt th nghim. Axit bo chui ngn (SCFA), l sn phm cui cng ca qu trnh ln men vi khun, c phn tch bng sc k lng hiu nng cao (HPLC). Vt liu. Mi ho cht u c mua t BDH (Poole, Dorset, Vng Quc Anh), ngoi tr cho MgSO 4 7H 2 O v l -cysteine HCl, thu c t Sigma (Poole, Dorset, Vng Quc Anh). Fructo-oligosaccharides (Raftilose P95) thu c t Orafti (Tienen, B), v tinh bt l t Sigma (Poole, Dorset, Vng Quc Anh).Ti co Hamlin cam sut phn chiu (phn trng ca v cam) c ly t mt b x l nc tri cy h cam qut Florida thng mi v ng lnh ngay lp tc n -20 C cho n khi s dng. Khai thc oligosaccharides pectic. Orange sut phn chiu c s dng trch xut cc oligosaccharides pectic. POS sau c iu tr bng cch lc nano loi b nitrat vt qu ( 14 ). Orange sut phn chiu c pha trn vi mt trng lng bng nhau ca nc kh Ion ha v sau lm t xay bng cch s dng mt b x l thc phm m hnh Comitrol 1700 (Urschel phng th nghim, Valparaiso, IN) dn tt vi mt phn b kch thc ht trung bnh di 0,5 mm.Cc mt t bn sut phn chiu (9 kg) c cha 4,5 sut phn chiu kg c thm vo mt chic thng cha 57 lt dung dch HNO 3 (pH 1.5) cung cp cho mt nng 1 g peel/12.5 ml HNO 3 , v c iu chnh pH n 1,5 tp trung HNO 3 . B nui cng c sau c khuy trong 30 pht, v bn c bm vo mt thanh trng (MicroThermics HTSL Lab-25, Raleigh, NC) cu hnh vi 7,75 mm, bn trong c ng knh ng trao i nhit (nng khi lng 400 ml, 400 ml gi, lm mt khi lng 400 ml). Thanh trng c iu chnh n mt nhit nng 120 C, p lc tr li ca 310 n 380 kPa ( ngn chn si), v t l lu lng t 1.200 ml / pht vi nc trc khi bm bn. Sau khi chuyn i bn, nhit u vo hi nc c tng ln mt cht cho bn nhit 120 C. Nh , bn

sau c un nng n 120 C trong vng 20 giy, c t chc nhit 20 giy, v sau lm lnh n 14 C trong vng 20 giy. Nhit th trong lp rp ng gi t hn 4 C. Bn c bm qua mt hon thin tuabin Pony (Mecat M, Orlando, FL), n v lc c hc lin tc vi mt mn hnh mt li 53 micron, trong d lng v khng ha tan c tch ra t cc phng tin khai thc. D lng v khng ha tan c ra sch bng cch s dng 1 lng nc v kch ng trong 30 pht.X nc c thng qua mt ln na thng qua vic hon thin turbo, ni cht lng c thu thp v d lng khng ha tan c loi b. Isopropyl alcohol (IPA) sau c thm vo chit xut cht lng a nng ln n 70% IPA, v gii php c lu tr trong 12 h ti 5 C kt ta pectin. Gii php ny sau c bm qua b lc turbo tch kt ta v kt ta khng tan trong cn c ra sch hai ln na vi 2 phn trn mt n v trng lng ca IPA 70% v mt ln vi 100% IPA. Cc phn khng tan trong cn sau sy kh trong l chn khng. Ni trn mt ha tan trong ru c trung ha vi dung dch NaOH pH gia 6.5 v 7.5, v IPA bc hi 35 C di p lc gim bng cch s dng Buchi (Brinkman, Westbury, NY) thit b bay hi quay 20 lt. Ni trn mt dung dch nc c ly tm 9639 x g loi b mt kt ta my hnh thnh sau khi iu chnh pH. Ni trn mt c vo mt C 18 ct chit pha rn (Altex, Harbor City, CA) tch cc cc t cc phn phn on khng cc. Cc phn cc c cha POS c ng kh, v cc phn khng cc c eluted vi ethanol. Lc nano oligosaccharides pectic. ngi thu thu phn cc, diafiltration lin tc c thc hin bng cch s dng 1.000 phn t trng lng ct DS-5-DL mng (Osmonics Desal). Cc phn cc (60 g) c ha tan trong H 2 O (2 lt) cung cp cho hm lng ng tng cng 3%. Gii php ny c lu hnh trong h thng lc trong 10 pht trc khi nng thc n ban u c xc nh. Hai song song cc t bo lc p sut cao c s dng, vi tng din tch mng 81 cm 2 . u vo dng chy ca nc trong bn cha thc n chn nui c kt hp vi tc dng chy trn ngp nh vy m khi lng khng thay i. p sut v nhit c thit lp ti 20 10 6 Pa v 60 C, tng ng. Thm nhp vo cc php o thng lng v retentate v thm nhp vo mu c thc hin mi h 2. Th nghim c hon thnh sau 6 lt tch ly trn ngp thu c. Retentate c cha v cam POS c ng kh. Trc v sau khi lc nano, ni dung nitrat ca mu c xc nh bng sc k ion bng cch s dng mt Dionex AS14A ct, mt ct bo v AG14A,

v mM 3,5 Na 2 CO 3 -1,0 mM NaHCO3 eluent tc 1.2 dng chy ml / pht. Hiu sut cao sc k trao i anion (HPAEC) Orange v POS c tch ra hn 100 min 25 500 oxalat kali mM phi tuyn, pH 6, gradient bng cch s dng mt ct PA1 CarboPac (4 250 mm; Dionex Corp) v mt CarboPac PA bo v ct trn mt h thng Dionex 4500i ( 18 , 19 ). Polygalacturonic axit thy phn trong mt ni hp 40 pht 125 C trong dung dch axt nitric. Tiu chun oligosaccharide c c lp bi d bi HPLC ( 17 , 20 , 21 ) hoc thu c t Sigma. Xt nghim carbohydrate. Cc ni dung carbohydrate tng s POS c xc nh theo phng php axit phenolsulfuric ( 6 ). Hm lng acid galacturonic c xc nh bng mt phng php so mu ( 8 ), vi mt s sa i ( 48 ). Phn tch tinh bt c thc hin bng cch s dng cc Megazyme (Bray, Ireland) kit phn tch tng tinh bt. B ti liu ny s dng chu nhit amylase v amyloglucosidase cho thy phn tinh bt v ng glucose oxidase / peroxidase pht hin glucose. Monosaccharide phn tch. Methanolysis POS c thc hin bng cch thm 100 acetate methyl ml v 400 ml 1,5 M HCl trong methanol mu 1-mg (sy kh 50 C l chn khng) v un nng 85 C trong 17 h. Sau khi cc mu c lm lnh ti nhit phng, 5 git n butanol c b sung v cc mu bc hi n kh theo mt dng kh nit. thy phn cc glycosides methyl hnh thnh bi methanolysis, 0,5 ml 2 M trifluoroacetic axit c b sung, v cc mu c gia nhit 121 C trong 1 gi. Cc mu c lm lnh ti nhit phng v bc hi n kh, v 0,5 ml methanol c thm vo v bc hi n kh ba ln.Nc c thm vo mi mu, v h c lc (0,25 micromet) trc khi n tim. Inositol c thm vo mi mu trc khi methanolysis nh mt tiu chun ni b. Thy phn c phn tch ni dung monosaccharide trung lp bng cch pht hin amperometric HPAEC-xung (PAD) bng cch s dng mt Dionex ICS2500 h thng bao gm mt CarboPac PA20 ct v ct bo v, 50 EG eluent my pht in m sn xut KOH 10 isocratic mM in thoi di ng giai on, lin tc ti sinh ct anion ci by, mt bc s 50 my bm hot ng mc 0,5 ml / min, mt ED50 in my d s dng cc dng sng tim nng tng gp bn ln, v mt mu t ng AS50 vi mt ngn nhit (30 C ct nng). Ni dung monosaccharide c tnh axit c xc nh vi mt h thng Dionex500 DX, trong bao gm mt my bm gradient GP50 (0,5 ml / pht), mt ct CarboPac PA20 ct v bo v, mt my d ED40 in (in cc lm vic vng v in cc tham chiu pH), mt LC25 sc k l (30 C), kh nn PC10

iu khin (postcolumn Ngoi ra 500 mM NaOH), v mt mu t ng AS3500. Giai on di ng bao gm ca mM isocratic 10 CH 3 COONa, 1 mM NaOH eluant cho 10 pht, v sau mt gradient tuyn tnh ca 100 n 130 mM CH 3 COONa trong 100 mM NaOH sau 20 min. Phn l nn vn ha qu trnh ln men. Bn tu ln men thy tinh 280 ml c lp y vi 180 ml ca mt mi trng v trng prereduced c cha 2 g / nc peptone lt, 2 g / lt chit xut t nm men, 0,1 g / lt NaCl, 0,04 g / lt K 2 HPO 4 , 0,04 g / lt KH 2 PO 4 , 0,01 g / lt MgSO 4 7H 2 O, 0,01 g / lt CaCl 2 2H 2O, 2 g / lt NaHCO 3 , 0,5 g / lt l -cysteine HCl, 0,5 g / lt mt mui, 4 ml resazurin 0,05 g / lt, 10 ml vitamin K 1 , 2 ml Tween 80, v 10 ml hemin. Mi trng c iu chnh pH 7.0 v lin tc sparged vi oxy nit (15 ml / pht). 10% (wt / vol) phn bn t ba nh ti tr khe mnh c chun b bng cch s dng prereduced 0,1 M phosphate m nc mui sinh l (pH 7.0) v sau trn ln trong mt stomacher (m hnh 6041, Seward khoa hc, Vng quc Anh) cho 120. Mi tu c tim 20 ml bn v g 2 carbohydrate (POS, fructo-oligosaccharides, cam sut phn chiu, hoc tinh bt). Mu phn c ly t mt con c v hai tnh nguyn vin n c tui trung bnh l 24 nm ngi khng dng thuc khng sinh trong t nht 6 thng trc khi cung cp cc mu v nhng ngi khng c tin s ri lon tiu ha. Fermentors c duy tr 37 C, v mu bn sao c g b sau khi 0, 5, 10, v 24 gi m s vi khun v SCFA phn tch (46 , 29 ). Qu trnh ln men c chy trn ba dp ring bit vi nh ti tr khc nhau. Thay i hunh quang trong lai ti ch. trong qun th vi khun phn ngi c nh gi bi hunh quang lai ti ch vi cc u d 16S rRNA. Cc u d 16S rRNA, c th cho cc lp hc ch yu ca h vi sinh ng rut (Bacteroides, bifidobacteria,: Clostridium histolyticum , Eubacterium rectale v lactobacilli / enterococci), c sn xut v c dn nhn nh du hunh quang. Cc u d s dng Bif164 ( 26 ), Bac303 ( 28 ), Erec482 ( 10 ), His150 ( 10 ), v Lab158 ( 16 ). Cc axit nucleic vt 4'6-diamidino-2 phenylindole (DAPI) c s dng m tng s vi khun. Mu qu trnh ln men c pha long v c nh qua m trong Paraformaldehyde 4% 4 C trong mt t l 1:03 ca mu Paraformaldehyde 4% (vol / vol). Nhng t bo ny sau ra sch vi nc mui phosphate-m (0,1 M, pH 7,0), resuspended ml 150 phosphate m nc mui cng vi 150 ethanol ml, v c lu tr -20 C ti thiu l 1 gi. Treo t bo c sau thm vo hn hp lai bao gm 4,78 g / lt Tris-HCl (30 mM), 79,69 g / lt NaCl (1,36 M), 15 ml 10% sodium dodecyl sulfate gii php (pH 7.2), v 4 M HCl 15 ml v lc thng qua mt b lc 0,2 micron (Millipore, Watford, Vng Quc Anh). Lai to c thc hin nhit thch hp cho cc u d. Sau , hn hp lai chn khng lc

(Millipore, Watford, Vng Quc Anh), v b lc c t trn mt knh hin vi v kim tra bng cch s dng knh hin vi hunh quang, chng hn cc nhm vi khun c th c lit k ( 36 ). . Phn tch acid bo chui ngn mt millilliter mu c g b t cc fermentors vn ha hng lot v ly tm 13.000 g trong 5 pht, v 20 ml c tim vo mt h thng HPLC (LaChrom L7100; Merck, Poole, Vng Quc Anh) trang b mt my d ch s khc x v mt vi phun t ng. Cc ct c s dng l mt ion-loi tr Aminex HPX-87H ct (7,8 bng 300 mm; Bio-Rad, Watford, Vng Quc Anh) duy tr mc 50 C. Eluent l 0,005 mM H 2 SO 4 ti mt tc dng chy 0,6 ml / pht. D liu c thu thp bng cch s dng JCL6000 phn mm (Jones sc k, Wales, Vng Quc Anh). nh lng ca cc mu c thc hin bng cch s dng cc ng cong hiu chun axetic, axit propionic, butyric, lactic nng gia 0,5 v 100 mM, v kt qu c th hin trong mmol / lt ( 36 ). Cc th nghim c lp i lp li trong ba ln. Phn tch thng k. Trong trng hp s khc bit trong phng php iu tr c tm thy, sinh vin hai mu th nghim t c thc hin gia cc mu. S khc bit c coi l ng k khi P <0,05. trc Mc sau

KT QU
POS khai thc. khai thc thanh trng ca POS t v cam Hamlin sn xut 158,5 g ca phn nh ni trn mt cc t 4,542 kg t v (976,5 g, trng lng kh), m l mt nng sut 16%. tip tc thanh lc phn cc ni trn mt v loi b mui c th can thip vi cc nn vn ha hng lot phn, lc nano c thc hin.Lc nano gim ng k (nm ln) nitrat ni dung n mc ti thiu v gim trng lng phn 50%. V vy, nng sut cui cng ca POS t Hamlin v cam l 8%.Nng sut tng ng vi cht lng pectin cao c sn xut (d liu khng hin th). POS thnh phn. Orange v POS bao gm mt lot cc oligosaccharides (Hnh 1 ). Sc POS l rt tng t nh sc cho cc phn ni trn mt cc c cng b trc (xem hnh 2D trong ti liu tham kho 21 ), chng minh rng lc nano khng lm thay i h s c nhn POS oligosaccharide. POS thi gian lu gi oligosaccharide khng chnh xc ph hp vi nhng cc axit oligogalacturonic (Hnh 1 ), cho thy rng POS c mt cu trc rhamnogalacturonan. Khi POS c saponified vi KOH (11 pH, 30 pht) v

sau v hiu ha vi HCl, vic lu gi thi gian v chiu cao nh im ca mt s nh ni trong hnh 1 . chuyn, v mt lot cc nh ni nh gia 25 v 55 min c quan st (d liu khng hin th). Thi gian lu ca cc nh ni mi xut hin sau x phng ha cng khng ng vi thi gian lu gi axit oligogalacturonic. iu ny cho thy rng mt s d lng axit galacturonic trong POS c sa i, c th vi cc este methyl. POS chnh xc mc trng hp l khng r. Tuy nhin, k t khi chiu di chui oligosaccharide thng tng ln cng vi cc tng HPAEC-PAD thi gian lu, v cam POS phm vi chiu di chui (Hnh 1 ) l ln hn so vi phm vi chiu di chui POS I v II POS ( 22 ) c sn xut bi enzym thy phn t cao cy c mi pectin-methoxy-methoxy pectin to, tng ng ( 30 ).

Xem phin bn ln hn:


Trong trang ny Trong mt ca s mi Ti v nh slide PowerPoint

Hnh 1. HPAEC-PAD phn tch ca POS chit xut t v cam (A) so vi mt thy phn axit polygalacturonic (B). Cc mc trng hp tng ng vi tiu chun axit oligogalacturonic c ch ra trn cc nh ni. Thnh phn monosaccharide ca POS bao gm ch yu l glucose v arabinose, vi s tin thp hn ca galactose, axit galacturonic, xylose, rhamnose, v fucose (Bng 1 ). POS c mt ni dung axit galacturonic 13,0% ( 1,1%) v mt ni dung 56,6% ( 1,4%) tng s carbohydrate (glucose tiu chun) c xc nh bi cc xt nghim o mu. Nng cao ca hin glucose trong thy phn POS l do glucose min ph trong phn nh POS. Ni dung glucose ca POS l 18% v khng c iu tr -amylase v amyloglucosidase trong vic kho nghim tinh bt. S hin din ca glucose trong phn POS unhydrolyzed c xc nhn bi HPAEC-PAD bng cch s dng ct PA20 CarboPac. ng min ph c pht hin trong sut phn chiu mu da cam (7%) c s dng trong nghin cu ny. Thnh phn monosaccharide cn li l ph hp vi mt cu trc rhamnogalacturonan c cha axit galacturonic 03:01 / rhamnose t l. Bi v t l ny l 1:1, POS c th l mt s kt hp ca rhamnogalacturonan v xylogalacturonan trong t nhin ( 35 , 45 ). Mt s cc nh ni trong hnh 1 c th i din

oligosaccharides, arabinan hoc arabinogalactan. l in hnh chui bn rhamnogalacturonan. N cng c th l mt lng nh ca oligosaccharides xyloglucan c th c hin nay da trn cc thnh phn monosaccharide POS. Tuy nhin, da trn POS c chit xut v thc t rng khng c bng chng cho cello-oligosaccharides c mt trong HPAEC-PAD sc k , cc oligosaccharides xyloglucan c l ch c cc thnh phn ph, nu c tt c. Phn tch su hn s c thc hin v sau bo co xc nh cu trc chnh xc ca POS. Xem bng ny:

Trong ca s ny Trong mt ca s mi

BNG 1. POS monosaccharide thnh phn Nghin cu qu trnh ln men. Nhng thay i trong nhn dn s phn vi khun hin din trong cc nn vn ha hng lot c tm tt trong Bng 2 . C hai POS v FOS sn xut gia tng ng k trong bifidobacteria sau 24 h ln men vi POS v sau 5 h 10 vi FOS. Bacteroides dn s vn n nh cho ba trong bn mu th nghim, vi ngoi l ca tinh bt, ni mt gia tng ng k c tm thy sau 5 h ln men. Tng t nh vy, Clostridium histolyticum mc c duy tr vi mt con s gim sau 24 h ln men vi FOS. Tng ng k c nhn thy trong Eubacterium cc s rectale vi POS v FOS, v tng khng c ngha c xc nh vi cam sut phn chiu v tinh bt. Tng lactobacilli / enterococci cng nhn thy, nhng mt ln na, s gia tng ny l khng ng k. Xem bng ny:

Trong ca s ny Trong mt ca s mi

BNG 2. Qun th vi khun sau khi 0, 5, 10, v 24 gi vi oligosaccharides khc nhau Nng axit lactic, acetic, propionic cao tt c cc thi im th nghim vi POS nh l mt ngun carbohydrate (Bng 3 ). POS cho SCFA tng s cao nht ca tt c cc carbohydrate c th nghim ti 24 gi. Butyrate sn xut,

tuy nhin, tng ln sau 10 h v t v tr 24 gi, trng vi cao nht rectale Eubacteriums. FOS dn n tng ng k nng axit axetic, propionic, butyric sau 10 h v acid lactic sau 24 h. Cam sut phn chiu ln men sn xut tng axit lactic, acetic, propionic lc 5 h, v nng hp cht butirat t v tr 10 h v vn cao ln n 24 h, mt ln na tng ng vi Eubacterium rectale . Sau 24 h ln men vi tinh bt, tng ng k trong axit propionic c nhn thy. Sau 10 gi, nng acid butyric tng v vn mc cao cho n 24 h. Xem bng ny:

Trong ca s ny Trong mt ca s mi

BNG 3. Nng ca axit hu c c sn xut trong qu trnh ln men ca mi ngun carbohydrate 0, 5, 10, v 24 gi iu kin ym kh pH 6,8 v 37 C 1 Prebiotic ch s (PI) Mt phng trnh nh lng c s dng gip phn tch qu trnh ln men prebiotic ( 33 ) cho POS hin th mt xu hng tng t c tm thy trong nghin cu trc y ( 29 ), tng vi thi gian (Bng 4 ), nh sau: Xem bng ny:

Trong ca s ny Trong mt ca s mi

BNG 4. PI POS, FOS, vim khp, v tinh bt sau khi 5, 10, v 24 gi ca qu trnh ln men k kh pH 6.8 v 37 C FOS hin th mt xu hng tng t, nhng t c gi tr cao hn. Vim khp hin th mt s im PI ban u cao, nhng iu ny gim vi thi gian. PI cho tinh bt c tm thy gim sau 10 h, c th l do mc tng ca Bacteroides v clostridia ti thi im ny. trc Mc sau

THO LUN
Hng lot th nghim nui cy c thc hin vi POS v vim khp, bi dng sn phm sn xut nc cam, v so snh tim nng prebiotic ca h chng li FOS v tinh bt l cht kim sot. Trong khi v cam POS v vim khp c cha glucose min ph, carbohydrate ny c khng bit l c tnh cht tin sinh hc. V vy, cc POS c ngun gc t v cam chu trch nhim cho cc gi tr PI cao. Trc y, enzym to ra POS c bo co l c tim nng l cht tin sinh hc ( 29), vi PI nng cao lin quan n nhng ngi ca pectin cha m ca h. Chn lc l khng, tuy nhin, nh nh du l c nhn thy vi FOS, c th l do vic duy tr Bacteroides v qun th vi khun Clostridium trong sut qu trnh ln men. C cht pectin c hin th duy tr mc ca tt c cc vi khun trong cc th nghim ln men trc ( 7 ). Do thay i trong cc nh ti tr phn, cc gi tr PI thu c cho enzym to ra POS ( 29 ) v PI POS gi tr thu c y khng th so snh trc tip. Tuy nhin, n c kh nng v cam POS PI s tip tc tng sau 24 h, nh c quan st cho enzym to ra POS ( 29 ). POS v cam cng tng dn s bifidobacterial sau ny trong qu trnh ln men so vi FOS, cho thy v cam POS sn xut ln men prebiotic bn vng hn. Mt s pha trn ca POS v FOS c th dn n con s cao bifidobacterial mt thi gian di ca thi gian ( 9 ). Vic s dng cc b trao i nhit ng khai thc lin tc ca pectin c ci ng c bo co trc ( 47 ). y l bo co u tin ca phng php sn xut pectin v POS t v cam. Trong khi sn lng ca POS l 8%, c tnh s b ca chng ti l n s khng t hn sn xut thng mi hn so vi pectin.POS l mt thnh phn quan trng ca cc ni trn mt cn li sau khi lng ma pectin. Ni trn mt y l mt vt liu ph thi vi mt nhu cu oxy sinh hc cao l rt tn km x l trong qu trnh sn xut pectin. V vy, POS c th l mt coproduct c th h thp chi ph sn xut pectin. Eubacterium rectale tng ln ng k vi oligosaccharides th nghim. Ch n ung cht x trong cc hnh thc ca pectin tri cy trc y c hin th tng cc s eubacterial ( 7 , 38 ). Ngoi ra, pectin vi cc mc khc nhau ca kt qu este ha khc nhau nng butyrate ( 5 ). Nhm ny sinh l quan trng ca vi khun sn xut s lng tng i ln ca butyrate ( 38 , 1 , 24 ). Sn xut Butyrate c cho l mang li li ch cho sc khe ( 4 ), vi qung co vai tr trong cng tc phng chng vim lot i trng ( 3 , 39 , 41 ) v ung th rut kt ( 15 , 34 ).

Trong kt lun, chng ti ch ra rng POS c th c s dng c hiu qu nh l mt prebiotic bi v s gia tng bifidobacteria v Eubacterium rectale s vi s gia tng tip theo trong nng hp cht butirat, a ra mt li ch sc khe c th gia tng. trc Mc sau

LI CM T
Chng ti cm n Andr Trng axit galacturonic v phn tch tng carbohydrate o mu v Michael Kurantz cho vic phn tch tinh bt. trc Mc sau

Ch Thch
Nhn c 19 thng T nm 2005. o Chp nhn 27 thng Tm nm 2005. * Tng ng vi tc gi. a ch gi th: Trng Khoa hc Sinh hc Thc phm, i hc Reading, PO Box 226, Whiteknights, c RG6 6AP, Vng quc Anh. in thoi: 44 (0) 118 3786726. Fax: 44 (0) 118 9310080. E-mail: rarastall@rdg.ac.uk .
o

Trc mc

THAM KHO
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k hiu sut trao i anion v pht hin amperometric xung. Anal. Biochem 184 : 200 -206. CrossRef Medline
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