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Metabolic engineering for biobutanol production:a review

生物丁醇代谢工程的研究进展



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ChineseJournalofBioprocessEngineering
Vol.11No.2
Mar.2013
doi:10.3969/j.issn.1672-3678.2013.02.009
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Metabolicengineeringforbiobutanolproduction:areview
DAIZongjie1,2,DONGHongjun2,ZHUYan2,ZHANGYanping2,LIYin2
(1.SchoolofLifeSciences,UniversityofScienceandTechnologyofChina,Hefei230026,China;
2.InstituteofMicrobiology,ChineseAcademyofSciences,Beijing100101,China)
Abstract:Butanolisanimportantbulkchemicalwithwideapplication.Butanolwasconsideredasanew
biofuelalternative.Duetotheenergyandenvironmentcrisis,biobasedbutanolproductionfrom
sustainablebiomasshasbeenpaidatentionworldwide.Metabolicengineeringasaneficienttoolhasbeen
usedwidelyforbiobutanolproductionwithdiferentmicroorganisms.Metabolicengineeringclassic
acetonebutanolethanol(ABE)fermentationwasasignificantdirectionforbeterbutanolproduction.The
rationaldesignbutanolproducingpathwayinmodelbacteriawasanotherchoiceforbutanolproduction.In
thisreview,theprogressesonmetabolicengineeringofbutanolproductioninrecentfiveyearswere
summarized,theproblemsinbutanolproductiontomeetthedemandsofindustrialproductionwerealso
discussed.
Keywords:biobutanol;metabolicengineering;metabolicpathway;biomanufacturing
  
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[1] DureP.Fermentativeproductionofbutanol:theacademic
perspective[J].CurOpinBiotechnol,2011,22(3):331336.
[2] MermelsteinLD,WelkerNE,BennetGN,etal.Expressionof
cloned homologous fermentative genes in Clostridium
acetobutylicumATCC824[J].NatBiotechnol,1992,10(2):190
195.
[3] NolingJ,BretonG,OmelchenkoMV,etal.Genomesequence
andcomparativeanalysisofthesolventproducingbacterium
Clostridiumacetobutylicum[J].JBacteriol,2001,183(16):
48234838.
[4] AtsumiS,CannAF,ConnorMR,etal.Metabolicengineeringof
Escherichiacolifor1butanolproduction[J].MetabEng,2008,
10(6):305311.
[5] LanEI,LiaoJC.Metabolicengineeringofcyanobacteriafor1
butanolproductionfromcarbondioxide[J].MetabEng,2011,13
(4):353363.
[6] LanEI,LiaoJC.ATPdrivesdirectphotosyntheticproductionof
1butanolin cyanobacteria[J].PNAS,2012,109(16):
60186023.
[7] BerezinaOV,ZakharovaNV,BrandtA,etal.Reconstructingthe
clostridialnbutanolmetabolicpathwayinLactobacilusbrevis
[J].ApplMicrobiolBiotechnol,2010,87(2):635646.
[8] SteenEJ,ChanR,PrasadN,etal.Metabolicengineeringof
Saccharomycescerevisiaefortheproductionofnbutanol[J].
MicrobCelFact,2008,7:36.
[9] NielsenDR,LeonardE,YoonSH,etal.Engineeringalternative
butanolproductionplatformsinheterologousbacteria[J]Metab
Eng,2009,11(4/5):262273.
[10] AtsumiS,HanaiT,LiaoJC.Nonfermentativepathwaysfor
synthesisofbranchedchainhigheralcoholsasbiofuels[J].
Nature,2008,451:8689.
[11] DelomonacoC,ClomburgJM,MilerEN,etal.Engineered
reversalofthebetaoxidationcycleforthesynthesisoffuelsand
chemicals[J].Nature,2011,476:355359.
[12] LehmannD,HonickeD,EhrenreichA,etal.Modifyingthe
productpaternofClostridiumacetobutylicum:physiologicalefects
ofdisruptingtheacetateandacetoneformationpathways[J].
ApplMicrobiolBiotechnol,2012,94:743754.
[13] JangYS,LeeJY,LeeJ,etal.Enhancedbutanolproduction
obtainedbyreinforcingthedirectbutanolformingroutein
Clostridiumacetobutylicum[J].mBio,2012,3(5):e0031412.
doi:101128/mBio.0031412.
[14] KuitW,MintonNP,LopezContrerasAM,etal.Disruptionof
theacetatekinase(ack)geneofClostridium acetobutylicum
resultsindelayed acetateproduction[J].ApplMicrobiol
Biotechnol,2012,94:729741.
[15] CooksleyCM,ZhangY,WangH,etal.Targetedmutagenesisof
the Clostridium acetobutylicum acetonebutanolethanol
fermentationpathway[J].MetabEng,2012,14:630641.
[16] LehmannD,RadomskiN,LütkeEverslohT.Newinsightsintothe
butyricacidmetabolismofClostridiumacetobutylicum[J].Appl
MicrobiolBiotechnol,2012,96:115.
[17] JiangY,XuC,DongF,etal.Disruptionoftheacetoacetate
decarboxylase gene in solventproducing Clostridium
36 
!

# àÑáa

&¢ˆ¢“)+N:;ÐM
acetobutylicumincreasesthebutanolratio[J].MetabEng,2009,
11(4/5):284291.
[18] BondWatsB B,BeleroseR J,ChangM C Y.Enzyme
mechanismasakineticcontrolelementfordesigningsynthetic
biofuelpathways[J].NatChemBiol,2011,7(4):222227.
[19] ShenCR,LanEI,DekishimaY,etal.Drivingforcesenable
hightiteranaerobic1butanolsynthesisinEscherichiacoli[J].
ApplEnvironMicrobiol,2011,77(9):29052915.
[20] ShenCR,LiaoJC.MetabolicengineeringofEscherichiacolifor
1butanoland1propanolproductionviatheketoacidpathways
[J].MetabEng,2008,10(6):312320.
[21] AtsumiS,LiaoJC.Directed evolution ofMethanococcus
jannaschicitramalatesynthaseforbosynthesisof1propanoland
1butanolbyEscherichiacoli[J].ApplEnvironMicrobiol,2008,
74(24):78027808.
[22] SteenEJ,ChanR,PrasadN,etal.Metabolicengineeringof
Saccharomycescerevisiaefortheproductionofnbutanol[J].
MicrobCelFact,2008,7:36.
[23] deBontJAM.Solventtolerantbacteriainbiocatalysis[J].
TrendsBiotechnol,1998,16(12):493499.
[24] KimBH,BelowsP,DataR,etal.Controlofcarbonandelectron
flowinClostridium acetobutylicum fermentations:utilizationof
carbonmonoxidetoinhibithydrogenproductionandtoenhance
butanolyields[J].ApplEnvironMicrobiol,1984,48(4):
764770.
[25] NakayamaS,KosakaT,HirakawaH,etal.Metabolicengineering
forsolventproductivitybydownregulationofthehydrogenasegene
clusterhupCBAinClostridiumsaccharoperbutylacetonicumstrain
N14[J].ApplMicrobiolBiotechnol,2008,78(3):483493.
[26] JonesSW,ParedesCJ,TracyB,etal.Thetranscriptional
programunderlyingthephysiologyofclostridialsporulation[J].
GenomeBiol,2008,9(7):R114.
[27] RenC,GuY,HuS,etal.Identificationandinactivationof
pleiotropicregulatorCcpA toeliminateglucoserepressionof
xyloseutilizationinClostridiumacetobutylicum[J].MetabEng,
2010,12(5):446454.
[28] XiaoH,GuY,NingY,etal.Confirmationandeliminationof
xylosemetabolismbotlenecksinglucosephosphoenolpyruvate
dependent phosphotransferase systemdeficient Clostridium
acetobutylicumforsimultaneousutilizationofglucose,xylose,and
arabinose[J].ApplEnviron Microbiol,2011,77(22):
78867895.
[29] XiaoH,LiZ,JiangY,etal.MetabolicengineeringofDxylose
pathwayinClostridiumbeijerinckitooptimizesolventproduction
fromxylosemotherliquid[J].MetabEng,2012,14(5):
569578.
[30] HenstraAM,SipmaJ,RinzemaA,etal.Microbiologyofsynthesis
gas fermentation for biofuel production[J].Cur Opin
Biotechnol,2007,18(3):200206.
[31] GrethleinAJ,WordenRM,JainM K,etal.Evidencefor
productionofnbutanolfromcarbonmonoxidebyButyribacterium
methylotrophicum[J].JFermentBioeng,1991,72(1):5860.
[32] LiouJSC,BalkwilD L,DrakeG R,etal.Clostridium
carboxidivoranssp.nov.,asolventproducingClostridiumisolated
fromanagriculturalsetlinglagoon,andreclassificationofthe
acetogenClostridiumscatologenesstrainSL1asClostridiumdrakei
sp.nov[J].IntJSystEvolMicrobiol,2005,55(5):20852091.
[33] KopkeM,HeldC,HujerS,etal.Clostridium ljungdahli
representsamicrobialproductionplatformbasedonsyngas[J].
PNAS,2010,107(29):1308713092.
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