OligosaccharideFormation

OligosaccharideFormation - _. _ :g-t—Oh—lt...

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Unformatted text preview: _. _ :g-t—Oh—lt _"—‘l __'—‘~T PM flmtinqfllflfll I594?! OLIGOSACCHARIDE FORMATION DURING HYDROLYSIS OF LACTOSE WITH SACCHAROMYCES LACTIS LACTASE (MAXILACT°}——PART 3: DIGESTIBILITY BY HUMAN lNTESTlNAL ENZYMES IN VITRO" A. BURVALL‘ N.-G, A5! & A. DARLOVIST Departmrnt of Applied Nutrition, Chemical Clntrt, University nj Lund. Lund. Sweden (Received: l5 July. I978) ABSTRACT Practically all the aligasacrliarlde: produced during enzymatic hydrolysi: oflactore with Saccharomyccs lac tis lactose nrej'ormea' l: y attachment of galactoryl residues in fl-(l —~ 6) linkages. The activity of human small-intestinal fl-galnctosidase on these substrates was less than 10% oft/19 activity an lactose which has its galactosyt moiety in a |tB-(t' —. 4) bond. Therefore. ingested diracchartdes will pass undigested through the small intestine into the large bowel. where bacterialdegradation takesplace. This explain: the gastrointestinal discomfort after allgosacchart‘de ingestion described in another paper (unpublished). INTRODUCTION Formation of oligosaccharides has previously been demonstrated to occur when lactose is hydrolysed with Sat‘chnromyccr lat-ti: 1actase(Maxilact®). This formation is due to the transglycosidasc activity of the enzyme and occurs particularly in solutions with a high concentration of lactosc (Burvall et at, 1977). The oiigosaccharidcs have been separated and structural analyses of the quantitatively most important sugars have bcen pcl‘formed (Asp e1 al..|1979]. The human small-intestinal lactose that hydrolyses dietary lactose has a rather strict substrate specificity (Asp 8: Dahlqvist. 1974) and intestinal discomfort. mainly flatulence. has been demenstrated to occur when healthy subjects are given milk containing large quantities (9 g) ofoligosaccharidcs formed during the hydrolysis of ' Part 1 ul‘tttis puperuppcarcd in Volt 4. No. 4.pp. 243-50: Part 2 appeared in Vol. 1ND. 2. pp. [47—53. I89 Ftl. Chem. 0303—81461‘8010005-0]h9l502'25 r0 Applied Science Publishers Ltd. England. [980 Printed in Great Britain 2'! ‘ilit (malt m I ylfilqllflf'? pen r:==_-r-. L5H I” A. IUIVALL. N36. m. a. mummy lactoseMsperal. unpublished dun) I ' . l n lhll H of {hue hnsglycuylnlian product: by lllllnll ouullsnrcannlnz mnumou ouulla HVDIDLYSIS as mums I91 "4’ "imam-zed m. «swam w “a [at ullylil or um um. gimme-oxidise (GLOX-novum. lubi All. rl mleu-aullal luau: whom_ Sweden) Ind plume with gallstone-dehydroan (Dnhlqvill. A 59. I971]. Blink: were pmpnmd will: Ill: urn: composition lam no! incubated, In million II: [III digestive brush horde: lm line hull-ml mall intestinal “mu Illa connin- m intracellular fl-placlnlidm Wilh unknwrn dilut‘m hm; The male: mun: [or only ghoul 57'. of the wul flpllclmidun filmy of Ill: Myriam-us med in lliil invutiplion will: helm: u Subllllll. my “Mum cm. hm‘ mninly m “=1 enlirnly he hydlblyled by Ill: nlmllllhrulzym (Ann .1 Dnhlqvill. I974). Th: hydmlylis of Dlilmhll‘ldfi as More Ilia ducrminnd in the WW (lle inhibllnr of [hen enzymu. rcMB. “Momma hem“: (obi-ind from KEBO AB. Stockholm. Mm) Thirly-llx millimlnm oflhin inhibitor um: diwlved in 5 lnlar0>l I NIDH. Tcn min-olives ul' nun [lock solution war: dilulnd with 5001mm unit-album: lohnion used I: described nblm. The final PCMB concenlrll'lon mined [(H mmcletl‘ltrnl inhibus Ih: inlmllular finplmlmidm almost mplcldy. Mlmfld Mmbnlians A plnlanpd incubalion with nous lnlnlinll alt-momma wu Also pufannzd will! "I dichrwl subnnles. Sammy-rm microlilrcs «if [M hulfer Subllrltewluliun dumbed llllovc wen: dilmad willl 2151mm: nl'nvl M Indium male“: pH 6 bufl'er. Tu Lllll dilulinll wu ll‘idlxl 300 .ulilru lllldihluad Inleflinnl humomue. The hydrolysll um followed by glues: Ind gunmen Innlysi: (u «scribed above) or lmnlllre umpla uteri allu- a. U2. I. 2 and all. MATE-nu AND moons Hum Mull-mutiny! enzyme prepamulln “samples”! Ind micron ‘ n11: ans; ul hum... "a. n a"; V V pull! no I pi ‘ ‘ “’3” null-Nd Full"! '“liifll Ell-5n. I:th and from within all: lie rl fit _ I I I Gilgamcdmn‘ab preparariru Tll: dill'erenl Random, 'A'.‘B'. ‘C Ind‘D' :l nt.|l979) were used I: subs letmuharldc: "I". Fm described in I prw-iml: plpeflflurml on A coal-incl! lriuochnridu 1nd I A,: fi-D—Gal-(l —- §)-fl-D-Gal-(l - él—flrD-GII-(l aq-n-Glc A,: fi-D-Gnl-(I ~o £)~3—n—Gal~[l a 61—0-th A,: fl-n—Gnl-(l ~6yfl-D—GIl-(l - syn-GI: Fraclinn ll :ornprind mni J ' ‘ u a “.0le n y the nmhande El: Mal-(l dermal“ Fraclicrl Ccomnined the dinmh-ridu: mm“ C .' - -Gnl- .. Jun 1: Gil-WI] ma “WWW/“"im‘m'fl'MW” m initlll men uf Iibcmiun of limos: and gala-ms: from 0-: diffuenl C,J [HS-61H] -o 6)-|>-Glc I (I «4)»D-Glc) hulitalnconumcd lhl dihechlridacl Ind CI Ind.inldlikion , a. me "mum comp-rad mm mm“ In Show“ in In”: l‘ The I TABLE I INnnL In: by NYDIDLY- w nu curl-ulnar majoruwh-nd,=xe:pllmwle no ' gull: I ' 2mm": :hydmlysil vim fi-nlu'naili';:n::h;{l - 5min” m be mm BED" All allm clnmhals used were Dr mlyl‘nl 51.1: mm, ‘ ' ;"I Ml Ana} :1] initial mainly mu; I Lllmm alumna-(Write {y . Tosn . - In pl! nu qf’laclm 1V [wwglllfslaffl Luigian ancient; bufl'a,pl-I H.9unuinin'lhe mmlmm I Sultan-lo Gnhrlm an... m:l'fl7§ul;lfl till/Jim lrfii 6' 2 I. m: D 2” w“ “wed “Nina al' the intestinal Imam Illo loo mud-mm 1 a : <5 ‘Jmo > ‘ ‘a 'mo mung , m mm) 8mm" m 5mm" “’“7‘ “l” omnirmmfl'ifmr 13:? I g ‘7”. 73 { 192 AV IIUKYALI.‘ ".43. “F. A. minnows-t liberstion of glucose and salmon {mm Janus: was arbitrarin set to 100.11, deem it hydmlyu’: oft-emu I!“ less than 57,. In fructinn D. committing approximately 80?, lei-ante. the liberatinn urging“ Ind pllcim WI: 70 Ind 'll 7... myocliwly or that in the lectnse control. 11; hydrniysis of the other oliguuochlride {flattens proceedud u I much rowan“ 1'hus.unfnclinn C the liberuion org!ch accurrad Il6 anlhe rum in lb: lasts. mnlml. The liberation of placing: from this Iranian mid I101 be mum saunter due to lactic" of unhydrulwed orignmlun'des (PMDIIIHy [all-cushion) with [he gllmton-dthydruaem my system Simificahl libcnu‘on nf'sallcwllwni obi-inn: both in Fraction A end in [twin . cont-int"; iii and leu‘uaocl'ulrides. respectively. n main tamponcnll The meat: thermiun regislefed in fraction B W]: on the borderh'ne of significninee TABLE] mm at cam-sen wmc um IL mu nun-rt PCMI was Am m tun-n In- “isle mew-u wminui :mn unu- HV‘ Mm —-—_______ um...” pf mun-magpie. In ,rr mi he: ammu- cmch ‘1 51:; ‘___‘__N_ bet-u: tan in: A 2 ‘2 s c 2 s 1 c d l D 1o w _ _ mosteompietely eboi'slznd b PC the Intrueeiluler fl-g: [Ictmldlsu which. in viva, pruhsbly can't-tot cutie hub: with Infifld alignsacchsrides if :1] seem rt; ‘ - > . wnnbl: forth: slum-h ' In and tetrusuehsndsr demunstntnd in the experiments wilIttautyld"‘<:mli’ly:‘anhe Prolonng inmban‘mi y then gal-emu mdmatml The hydmlyn‘sal'fncuon D lormoffreemononcelu dewwmundh “mew-"liberated more here Mu I slaw continuum OLIOOSACCILAKIDE mlMATION DURING minnows: ur “cross Slum “MINI” With” “Dir-Una ai__ ie—m .I-oog «a» war “mtmtedqlmfl WW 0 q 1 3 A. 5 B d | 2 3 IE 5 6 "my h Timej-t F‘. I, Llhlrllim 9151's“: Elicia“ nn minim math-lion uf III: dil'ml funicu- AID—D): I (A—A): CIA—m. tl—OI {as heImtO—ot Na PCMI nu sddsd in [his upenmwtl. ibis-lion at! both glucose Ind ulactase with I4 7; cf thc suglr as monaswchsrides mu 6h. From factions A and B pianos: was libasiets slawly. whereas no g’guifiauit iii-noun: of glucose was released went after a hours ni’incuhetinu. DISCUSSION In nntonccltltltcd milk mwheylhe formation nitunssiymsylslion products ts 10w but up to 2—1 yiilrc may occur under certain circumstances. During hydrolysis in minim mill. however. over )0 Zofthetuialsugnrmnlcnt mly be In the form aimlycosylntian productstkubctts in. Pettiuti. IOST). I‘h'n investinuon shows that these nligossechurides. formed during enzymic hydka of laclmewilh Sat'cilurnm’w: fartisluiasemannntbe hydrolysed u: [my Flflitllly impofllnl extent by the hum-n smull-tnlestimtl 11mm. The slow Ilum'on or phunse from the triv and letrssauhsridc-cnntlining rr-clions mutt-ted in the absence or PCMB seemed to be entirely due to inlneellulsr man without itan digestive functions A mliminlry study of the activin nf Mail-4:1. Ind humsn mteslinai B- thmidase on lame: and purified allolaclnsew-n-Gsi-(J -u 6)-D—G'|t:J has been Enid nut in am lebamory. Letting V be the init‘ni rate 51' hydrolysis for Ihc m" 5"!“- lhn rmiml’ [Vlmouiynlloisctosflwns about 2 For Mnxihctfl but kirltnmln fi-plutosidase it was well out tat limes linen The small ullolmou hytiuiysin; ulivily demanllralod in Ihe human iniulinsl hamnmnne was prdnbiy due to the inlrlsclllular fi-nlmlusidlsfi rather 1km the digestive mhntdu liens: y"! in: A Mun III- ' ,. .i'E I94 A. BURVALL. N.-G. ASP. A. DAHLQVIST ACKNOWLEDGEMENTS The Investigation was supported by tthwedish Nutrition Foundation and Svenska Mejcriernas Riksférbund (Milkfood AB). REFEREN CE Asr. N-G..‘Bunva_u, A. DAHLQVIST. A.. ngcm. P. h Lunnnyn. A. (I979). Oli BURVALL. A.. A52. N-G. & DAHLOVET. A. (I979). Olsgosaocharide formation during hydrolysis ul' halos: with Snccharomyce: lac-1i: lulu: (Maxilacl’W—Pan I: Quantitive aspects. Food Chemixrry. ‘, 243—50. DAHLQVIST. A. & A311. N-G. (I9TI). Analytical modems-Ir . 44, 654—7. Romans, H. R. & Pman. J. D. (1957). J. Agr. Fond C m.. 5. 1304. Samoan. M. 1. (I945). J. Biol. Chem. 160 61-9. ...
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This note was uploaded on 01/06/2012 for the course BIOL 1408 taught by Professor Markgarcia during the Fall '11 term at Collins.

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OligosaccharideFormation - _. _ :g-t—Oh—lt...

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