Citric acid production using Aspergillus niger by solid substrate fermentation

254 Pages·2011·6.19 MB·English

by  Lu, Minyuan.

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CITRAICCID P RODUCTUISOINNA GS PERGILLUS
NIGEBRYS OLIDS UBSTRAFTEER MENTATION
A thespirse senitnpe adr tfiulafill meonftt h ree quirement
for dtehger oefeD octoofrP hilosionpP hryo cess
andE nvironmTeencthanlo laotg y
MasseUyn ivseirty
MINYUANLU
1995
ToM yM other
Abstract
Thea imo ft hiwso rwka st oi nviegsatstoel siudbr sattfeem rentaftoicroir nti acc id
producutsiiAonspnge grilnlgiuesir n,a na ttetmopp rto vsiydset ematic ainndf onnation
anu nderstantdhipenr go ceIosnfsi l.te ixaperimenptesor rfwmeerdse e tloea cnt
appropsruibrasatttaeen do rngiasTmh.u ksu,m raaa nda s traiAnsp eorfg inlilguesr ,
YanNgo .w2e rfeuo ndfva ourafboclrie t arciipcdr oducwthiioplnoe,t awtaosa p oor
subrsatdtuet eoi tesx cesnsiritovgece onn tTehnfetnne .e ntatwieorcneas r roiueutdn der
varicoounsd itii.ioenn.os c,us liuzmmeo ,i stcuorneta ennpdta t ricsliez e souftb rstahtee
4
too ptimtihzepesa er amresIt.ne oculubme tswie1ze0eansn d16 0s poregsk /u4m0a ra,
mositucroeen nttbse tween 76%15 a napdna drc tlsiei zes b4e atnw6de enw ere
mm
optilf moacri tarciipcdr oudctiIotwn a.sf u ontdh attho er ganismu pn tuatkrebisye nt
penteratiinnttgoh s eu bsttrhauttshe fe,re mentahtaido na rdeilraetcitwo inttshhh ei p
avabillseau rfaaercaeT. h es olsiudtb rsatef uownadts o havpeo ttehnaetb iialtlio t y
overcomaed vteehrfeesfc eot fh igcho ncentorfma ettiiaoolnn Assd .d itoif1o 05nm glkg
susbtroafFt ee2+ 2,5 m g/ksgu brsatotefC u2+7,5 mgs/ukbrgsa totefZ n2+an 1d0 5m g/kg
susbtroaftM en2+ hsaldit glshyt iamtuolerffeyc tsc irotinac c ipdr oducrtaitothneh ra n
inhiitboerfeyfc tBsa.s oendt hoep timciozneddi ttihkoein nse,t tihcseso lsoiufdb strate
frementaitnfli aoswnke sr set udTihemeda .x imoubms erved grraatmveai,xm iemturmi c
obsersvpeedcr iafiatcen o dv erparlold ucotfci ivtiartciyipc dr d ouctwieorn1e5 . g /k.gh,
122m g/.gbiomaans0ds. .4gh8/ k.ghr,e stpievcely.
Tod evetlhosepo lsiudb stfrreamteeno tnap trcioesesx,p eriwmeernpete sr fonniend
diffenrtet ypoefsr eactionrcsl,u ad giansg- sfolluiiddb iezdea,d g as-liquid-solid
fluiizdebde da,r otadtriunamgn adp ackbeeddE .x cefpottr h pea ckbeedrd e actor,
these sysftuoenmtdso b euw nesrtuea ibfloer ftnnehenet atdiuoteno h, a rcsohn ditions
ofa briaosnfi,rc t,il oonwm oistsuurpep olryc ,o mbinattihoenfssae tc oorfTs h.e
fernmteatiinto hne pabcerkde eadc wtaosor p timwiiztrehed s pteoac itflr o wr atbee,d
loadainnpdga rtsiiczBleae.s oendt hese opctoinmdiizttehikdeoi nnse,wt eircsest u died,
ii
andi twa sd etermtihnatethdf enn e entatailolno mwuecdhh i ghreasrto efc irtiacc id
productthiaonn o wbesreeri vnfe ldka ssi,. aem .a ximoubms ergvreadv imreattorefi c
19.g /k.ghan adn o verparlold ucotf0i .vi8gt2ly k. ghI.na na tetmpttou ndersmtaasnsd
and hteraantis nft ehre ssoulbrisadttf nneee nttaioenx,p eriwmeernceto sn duicnta e d
multyie-prla ackbeedrd e actor. bHeocwaoeufvts ehecre o, m pleoxfmi atstysr aenris nf
solsiudtb rsaftnneee nttaiotnh,e underosftt haainssdp ieincntgt h pirseo scwsa sra ther
limiNteevde.r thelmeusllsta,iy -petarhc ekb eeddr eacitmoprr otvheed mtarsasn sfer
coindsaebrlcyo pmarweidt thhs ei nglep alcakbyeeeddr wtihteh sbaemldeo adainndg ,
allopwreedc miesaese umrentt hgoerf a difeonrt sgc aistearsci,ibc di ,o maasnsd rhcs.t a
Ther esuslutgsg tehsattth meu lti-plaacykbeeerddr eacitsao sru aibtlree acftoorr
fruthienrv estiagnadht aitsoh npeso ,s siobfib leiiutnsyge fdo lra rsgcea plreo duction
ofc irtiacc iidns olsiudb stfrnenaetnet ation.
Thissy steimnaevtsitci goafst oilosinud b strfanetnee ntaftociriro tinac c ipdr oduction,
whicihst hfier srteo prtepdr,o viddeetsa iinlfeodnn aatniduo nnd erstoafnt dhiinsg
frementatteicohnn ology.
III
Acknolwedgements
Thea uthwoirs hteoas c knowltehfdelog leo wpienogp le:
Chiseufp ervAisssootrceP,i r aeosfsDorrI.,a Sn. Maddoxr efmoabrrl keha ignsuc ei da
ansdu pervHiisesin ocno.u ragpeamteinaetnn,edc n et hustiharsomu gthhcoeou utr osfe
thPihs. sDt.u wdaysg reaatplpyr eciated.
SuperviAssosrostc,ePi raeosfsoDrrJ,.o hDn. B rooks Parneodsf soDrrS., . MR.a o
BhamidairmriH,e atodhf e Dtehpera tmeonfPt or cessE navnidr onTmeecnhtnaoll ogy,
fotrh esiurp ervainsidin oetnsei trnt hpirsjo ect.
PreosfsRo.Lr.E artlhefe,ro meHre ad oDfe ptahreto mfPe rnotc aensEdsn vireonntmal
Technofloohrgi eysn cuoramgeentth rougthhpoeru jote ct.
Assioactee sPsrBooorfbC honDgr,J. o hMna wsoann Ddr B.ri aWni lkefsroot nh eir
guidaanncsdeu ppiontr htpi rsjo cet.
MrG.r ahMaamn derasnoMdnr R .o sDsa vifeotsrh eiro rastnu kdpi pndndeusrsit nhge
study.
MrJ.o hAnl geMrrB,. r ucCeo llainnMdsr D.o nM clean fwoirl latinhnedegx i cre llent
assiswtiatnahclt elh ree latteecdh nmiactatalen rldsa boreaqtouriyp mfbaernitc ation
requiretmheaanrtto dssu er itnhgpi rsjo ect.
MrJ.o hSny keMsr,sA nn-maJraiceka snoMdnr M.i kSeha ayafmot rh eliarb oratory
support.
MrM.i kSet evfeohnri asv ailatbosi ulpipttlhryyee qucihreemdi caltsh pidrsjuo ercitn.g
iv
Mr. WayMnael lefto rh iss uppoirntt her elatceodm putweorr .k
TheN ew ZealaMnidn istorfEy x ternaRle latiaonndsT radfeo rt hef inancsiuaplp oorft
thisst udy.
Mrs.N. Arnott & Mr.R .Arn ott,M rsB.. Willia&m Msr .A. WilliaMmrs.s, B . Sanson
& Mr. T.S ansofno rt hekiir ndneasnsdf rienid.ps h
Felloowfst heA ssocioaftC eh ineSsteu denotfMs a sseUyn iversFietnygY, u ,H ailong
Wanga ndL iO uyanfgo rt heiars sistaanncdke i ncdo ndseiration.
Postgradfuealtleo Mwss.,T aniNag apoM,r .D aviOdl dfieMlsd.P, i nthMiutnag karndee,
Ms.S irichLoema ngoMns,.B ongkoNto pponM,r .E dgaRru shizfhoasr h aritnhge tiirm e
andf riendship.
Chinefsrei enZdhsa,n gL i&n HuitiMnag, H ongC hen& XingjuXnu ,P atriXcika oP ing
Li& SusaQni ngS u,J iaPne ngG u & PeiZ hongX,i aoP inPga n& LianLgiK ongM,s .
Wen Luoa ndM r. ShirninF u fort heifrri endsshhiapr,i ng ktiinmden,e asnsds upport.
Theikri ncdo nsideraotnim oen,ws i llianngda vailaboiflh ietlaypr ed eeplayp preci.a ted
Littflrei enAdsst,e Yri LinJ,o hnL inaCnh ena ndJ ieGsui fors haritnhge niari vtei me.
My Fathesri,s taenrdb rothfeortrsh eciorn stasnutp poarntde ncouragetmhernotu ghout
thisst udy.
My father-iann-dlm aowt her-inf-olrta hwe siurp poarntd e ncouragement.
My wifJei nWga ngf ohr ert ypianngd a dvisionntg h et hesainsd,f ohre rl ovep,a tience
ande ncouragement.
v
Contents
Abstract i
. . .. . .. . . . . . . .. . . . . . . . . .. . .. . . . .. . . . . . . . . . .. . . . . . . . . .
Acknowledegnetms. .. ... .............. . . . . . . .. ............ .i.i.i
.
Conetnts . .. . . . . . ...... . .. . . v
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
LisotfF igure.s . . . . . . . . . . . . . . .. .. x .i i. . . . . . . . . . . . . . . . . . . . . .
. . . .
LisotfT able.s . . . . . . . . . . . . . . . ... x .v ii. . . . . . . . . . . . . . . . . . . . . . . . .
.
Nomencla.t u. re. . . . . . . . . . . . . . . .x i.x . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Chapte1r Introduction . . . . ... .. . 1
. . . . . . . . . . . . .. . . . . . . . . . . . . . .
Chapt2er LiteraRteuvriee w.. .... ... . . .. . .... .. 2
. . . . . . . . . . . . . .
21. Introduc.t i.o n. . . . . . . . ... 2 . . . . . . . . . . . . . . . . . . . .
.
2.2 HistoorfyC itrAicci dP roductio.n . . . .. ..2 . . . . . . . . .
. .
2.3 Organisms. ... . . .... . ... ... .3
. . . . . . . . . . . . . . . . .
2.4 NutritiSotnaatlou fst heM edium . . .. . . . 4
. . . . . . . . . . . .
2.14 .Carbohydrat.e. . ....... .... . .5
. . . . . . . . . . . .
2.4.Ni2t rogaenndp hosphat.e . . . ... ... ...6 .. .. . . . .
.
2.4.Me3t ailo ns. . . ......... .. . ... 7
. . . . . . . . . . . .
2.4.Ot4h eard diti.v. e s . . . . . . .. .8. . . . . . . . . . . . . . . .
2.5E nvironmeFnatcatlo .r s. . . . . . . .. .9. . . . . . . . . . . . . . .
.
25..1pH ... . . .... ... .. . 9
. . . . . . . . .. . . . . . . . . . .
2.25 .Temperat.ur.e. ................ ......... 10
2.5.Ae3r ation . . ....... . . 10
. . . . . . . . . . . . . . . . . . .
2.45 .Othefra ctor.s . .. . ... .. . . .1 1
. . . . . . . . . . . . . .
2.6 FundmaentaBli ochemiostfCr iyt rAicci d
Prodcution. .. . ............ 1 1
. . . . . . . . . . . . . . . . . .
2.7 FennentatTieocnh nolofgioeCrsi trAicci d
Producti.on . . ........ . .. ......1 4
. . . . . . . . . . . . . .
2.17 .Introduc.t.i.o..n.. ......... ......... . .1.4
2.7.L2iq usiudr fafceenn entat.io..n. ... ..... .. 14
. . .
2.7.Su3b mergfeedr mentat.i..o.n. .... . .. .. 16
. . . . .
VI
2.7S.o4l isdu bstrfaetrem enta.t .io n.. .. . . ... 1. 8 . . . . . .
2.8 TheG enerSaoll iSdu bstrFaetrem entat..i.o.n. .. ..... 20
2.18 .Introduc.t.i..o.n.......................2 0
2.8.F2il amentfouunsgi in s olisdu bstrate
fermteanti.o .n . . . . . . . ... 20. . . . . . . . . . . . . . . .
2.8.S3ub sttre.a............................. 21
2.48 .Environmefnatcatlo. r..s. ............ ..... 23
2.481.. Moistucroen te.n.t. ........... 23
2.48..2 pH ........................ 24
2.48..3 Temperatu..r.e. .............2 4
2.58 .Kinetic.s.............................. 26
2.8.R6ea ctofrosrs olisdu bstrfaetrem enta.t.i.o.n. ... 27
2.9 Summar.y. ..... ..........................,. 3 2
Chapt3er MateriaanlsdM ethods ............................. 33
31. Materi.a l.s . . . . . . . . . .. .3.3 . . . . . . . . . . . . . . . . . . . .
31..1Mi crboiologimceadli. a . . . . . . ...3 3 . . . . . . . . . . .
31.2. Chemical.s. ... ..................... 33
. . .
31.3. Raw substr.at..e.s. ..... . . . . ... 33 . . . . . . . . .
31.4. Gase.s. ........................... .. 33
.
31.5. Organi.s.m............................ 35
3.2 MediaS teriliz.a.t.i.o.n. . .................3. 6
. . .
3.3 CleanionfgG lasswa.r .e . . . . . . .. .3.6 . . . . . . . . . . . . . .
34. AvoidanocfeW allG rowtihn F laskasn dR eacto.r..s. .. 36
3.5 AnalytiMceatlh od.s . . . . . . . . ... 36 . . . . . . . . . . . . . . . .
35..1p H measurem.e n..t . . . . . . ...3 6 . . . . . . . . . . . . .
3.5.S2po rceo un.t. ..... .................... .36
3.5.T3re atmeonfst a mple..s. ................. 37
.
3.5..13 Treatmepnrto cefsosrr aw
substr.a.t..e. .............. 37
.
3.35..2 Treatmepnrto cefsosr
fermentastaimopnl e.s. ........3 8
.
Contents vii
3.45 .Determinaotfif ounn gal bio.ma.ss.. ........ 3.8
3.5.A5na lysoifss t arc.h . . . . . . ... 3.8 . . . . . . . . . . . . .
35..6D eterminaotfig olnu coamylaacstei vity
andf regel uco.s .e . . . . . . ... 3.9 . . . . . . . . . . . . .
3.75 .Determinaotfic oint raicci .d . . . ....... ..... 4.0
3.5.D8et erminaotfic oanr bo.n ............... .. .41
3.95 .Determinaotfic oanr bodni oxi.d .e . . .. ..4 1. . . . . .
3.150.D eterminaotfio oxny ge.n . . . . .. ..4 2 . . . . . . . . . .
3.151.De terminaotfit oenm perat. u.r e. . . .. .4.3 . . . . . . .
35..1M2ea suremeonfat i flro wr at.e . . . .. ..4 3 . . . . . . . .
3.153.M easuremeonfat i prr essu.re.. . . ........... 43
3.6 CultuCroen ditio.ns. . .......... ..............43 .
3..61P reparaotfis ounb stra.t e .. . . . .. ..4 3 . . . . . . . . . .
36..2P reparaotfii onno culfuomrf ermentat.i .o ns.. ..4 5 .
3.6.F3la scku ltur.e. . . .... .. .. ...........4.5. . .
36..4T ubec ultu.r e . . . . . . . .. .4.5 . . . . . . . . . . . . . . . .
3.6.G5as -soflluiidd izbeeddr eactcourl tu.r .e . .. .4. 6 . . .
36..6L iquid-solfildu-igdaisbz eedrd e actcourl tu.r e .. .4.9
36..7R otatidnrgu mr eactcourl tu.r e . . . .. ..4 9 . . . . . . . .
3.6.P8ac kebde dr eactcourl tu.r e . . . . .. .5.3 . . . . . . . . .
36..9M ulti-lpaaycekre bde dr eactcourl tu.r .e . .. ..5 6 . . .
3.7 DiscussoifoM ne thod.s. ....... .. .. .......... 59
. .
3.17 .Organi.s.m............................5 9
37..2R aws ubstr.a .t e. . . . . . .. ..5 9 . . . . . . . . . . . . . . .
3.7.T3re atmeonfst a mple.s. ...... ........... .5.9
37.4. Determinaotfif ounn gablio mas.s ... . ......5..9
3.7.D5et erminaotfic oanr bodni oxiudsei n0g. M2 NaOH
soluti.o n. . . . . . . . .. .6.0 . . . . . . . . . . . . . . . . . .
3.7.G6as -soflluiidd izbeeddr eactcourl tu.r .e . .. .6.0 . . .
3.7.L7iq uid-solfliudi-dgiazsbe eddr eactcourl tu.r .e. . .6 1
3.7.P8ac kebde dr eactcourl tu.r e . . . . .. .6. 1 . . . . . . . . .
37..9M ulti-lpaaycekre bde dr eactcourl tu. r.e . .. ..6 1. . .