The Pilot Scale Production Of Fructo-oligosaccharides And The Design Of The Special Equipment

This dissertation includes the researches on the enzymatic properties of P -fructosyttransferase (FTS) from Aspergdlus nigerASOl ,the optimization of produang fructo-oligosaccharides (FOS) conditions by FTS, the processing of producing FOS in pilot scale of 1 ton per batch and the special equipment needed in the pilot scale factory.1 Selection of microorganismsAn Aspergillus niger ASO1 producing fructosyltransferase (FTS) was selected as the most suitable strain for FOS production from an Aspergillus niger,, an Aspetgillus oryzae and an Arthrobacter. The optimization of fermentation conditions by cross expenments are : sucrose 5%,pepton I .0o,malt juice (based on solid content) 0.7%, CMC 0.5% NaCI 0.3%.The enzyme activity of this Aspergillus niger is 12.586 u/mI after 72 hours incubation at 30C, but the best Ut/Uh (Ut :Transfer fructose activity of P -FTS;Uh:Hydrolysis activity of Ff5) ,is 13.05 after 24 hours incubation at 30C, the best enzyme productivity is 50.33 mglml, all the optimization of pH is 6.0.2.Observation of the Aspergillus nigerASolMicroscope and electron microscope ware used to obsetve the Asper~iIIus niger ASO1 ,the strain having the typical properties of Aspergillus niger. The black spore which covered the surface of the cultivate materials can be seen after 3 days incubation at 30 ~C ,the most suitable pH=7.0.3. oghbo Of cutore coodbons(1) Sucrose me the wt M catw,a ~ itor Tbe ompnicntw is bether than impanto nben.(2) The main bors irkat the m pbowton we X1 (sro %), X2 (optan edmp,Xi(MgSO4 ed FeC6o/o), X4 6ncuboon hme hrs) by the atNr wtmech.X4 is the M iW forr, th6 for in tUms X2X1,X3.(3) the otl edued by ~mente isY=53. 483+2. 792X,+l. l17Xz+1. 358X,+5. 342X.+2. 123X,'ro. 788X,X,+2. 085X2"1. 763XA+l. 910X'+1. 898L' Y= W peUCtjOnthe retw hbh sura % and bo incubatjon bme can in-- opprOduedon in epement -sion.(4) the resuha of oghmbeon Of cuhare conddions are that suerese 6.0%, PePtOn0.3s%,ng ng MgSo4 0.0se%, rwt 0.the%,.The epe pbouop of thisAop. was more than 100mgjmI ther 67 hr incutalon at 30C, pH=7.0.4. Iffimobtwn --is aod pboUCtjon Of high-PUty FOS by the immoblbo besthe lmmobjlbon material edum atoinate was the by tJsiro bePtimbeonsatUwt wtnd twn, the ds of th6 -- of tw odl ed ndumaIginate is 1;1 (Whote odlf 2% SOdium abhate). Th6 a~ of the imndIed mis 851 Uto gu.5% Of the crube epe aeq. und6 upper edbons, the haffdime Ofthe immoblwtW is 273.5 hrs. W6 pboued 43.5% FOS C/o) ther 48hrsreabo We 30t, 4% imrnodIbe m eded' be immodbe beds can beued 144hrs toeq tO ped FOS (42.5%, edhrs) and (41 .2%,144hrs).the contains of themateriais US6d tO Pboudng FOS wefe sure (50O/), MgSO. 0mp5. Studbe on the bendon pe Of immodIbehon FTS tO pboudng FOS inaghatjon trintOrThe peudro Of FOS SyrUp in 30L agboon ferentor was StUdied before th6 pilOtscate Wmeot' The Wrimenta1 resuha shwt that the mOSt sUhabe reaofoncondthon is 50% the conedon Of SUgar, 5% the immOdIbe bedS, 54hrs thereaofon hrne, thus the FOS(o/0) can be up tO 46es. be ch of be fermentalionare \ bopeI'atUre (30'C). pH(7.0). eqhatjOn pe (200rPm) \ Veoplate quatw(0~24hr rt 1: 0.5 wtin,24we hf,1: 1wtin, 48 hcr21 hr: 12VNAnin).W6 aISOStUded on the fetwnthen pe in 1000L Agbon bor The crossMboat dedn (33) were ued to drimpe the bo m6dium, aod theresuha Shwt that the moSt suhabe wton wtum cobe Of 6%sUgar,0.8%pePtOn,0.9O/ matt jube, 7.7OAs edkl Of the eretw edium. AjrCOchon we or24hr 1: 0.5wtin,24we hr1: 1~in, 48 hr ~ end 1:1.2VNhoin,W 5% inocolare size, 30C,120dmin apitatiop pe, 68hrs --, theeope aoo was aboUt 1000uI. We Pndued the 50.9%. FOS at conddions Of 5%the immob1bo m, 30'C, 48hrs.6. The anaI~ Of sugar comPOedon inThe coot Of the indiwtuaI boons Of hoedbataridesFOS) werecompered by hbh permance I'ud ChromatOgraPhy and gas chromatDgwty. -rhetwO medOds are aII amithe and suhabIe fbr atne anaWs, but the shOrtCOmes arethat the equiPment of HPLC is very exsopsbe and be PretIeatmat Of the samghe is...