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- 歷史
Freefattyacidsareusefulassourcemate
Freefattyacidsareusefulassourcematerialsfor
theproductionofbiodieselfuelandvarious
chemicalssuchaspharmaceuticalsanddietary
supplements.Previously,weattaineda9.2-fold
increaseinfreefattyacidproductivitybydisrupting
apredictedacyl-CoAsynthetasegene(faaA,
AO090011000642)inAspergillusoryzae.Inthis
study,weachievedfurtherincreaseintheproductiv-
itybyoverexpressingapredictedtransketolasegene
ofthepentosephosphatepathwayinthefaaAdis-
ruptant.TheA.oryzaegenomeispredictedtohave
threetransketolasegenesandoverexpressionof
AO090023000345,oneofthethreegenes,resultedin
phenotypicchangeandfurtherincrease(corre-
spondingtoanincreasedproductionof0.38mmol/g
drycellweight)infreefattyacidsat1.4-fold
comparedtothefaaAdisruptant.Additionally,the
biomassofhyphaeincreasedat1.2-foldbytheover-
expression.Asaresult,freefattyacidproduction
yieldperliterofliquidcultureincreasedat1.7-fold
bytheoverexpression.
Keywords:transketolasegeneoverexpression;
pentosephosphatepathway;freefatty
acid;productivity;Aspergillusoryzae
faaAdisruptant
Involvedintheincreasingglobaldemandoffossil
fuels,biofuelsgeneratedfromplantbiomasshavesig-
nificantlybeenfocusedduetolesseffectonglobal
environmentinthecarbonneutralcharacteristics.1,2)In
addition,somepharmaceuticalsanddietarysupple-
mentsproducedfromrarenaturalresourcesareexpen-
sive,andtheirproductionbytheheterologous
microbialbiosynthesisforloweringthepriceshasbeen
desired.3,4)
Microbiallipidproductionhasbeenresearchedin
cerevisiae,8−10)Yarrowialipolytica,11)etc.),filamentous
fattyacid(FFA),acylglycerol,fattyacidmethylester,
andalkanehavebeenthemajortargetsofthemicrobial
production.1)Amongthem,FFAcanbeusedassource
materialsofbiodieselfuelsandchemicals.1)
ThefilamentousfungusA.oryzaeiswellknownfor
highproductioncapabilityofhydrolasessuchasamy-
lase,protease,andlipase.17)Inaddition,A.oryzaeis
notonlyaproducerofsmallmoleculessuchasthe
kojicacid,18)butalsolistedasasafemicroorganism
fortheGRAS(i.e.GenerallyRecognizedasSafe)sta-
tusbytheUSFoodandDrugAdministration.19)Thus,
itimpliesA.oryzaeissuperiortotheotherfilamentous
fungiintheproductionofawiderangeofbiomole-
culesforindustrialapplication.Infact,ithasbeenused
forproducingsake(ricewine),shoyu(soysauce),and
miso(soybeanpaste)byconvertingstarch,proteins,
andlipidsderivedfromrice,wheat,andsoybean,to
manyvarietiesofsmallmoleculesthroughthecatabo-
lismandtheassimilation.
Inourresearch,A.oryzaehasbeenusedforaccom-
plishinghighproductionoflipidssuchasFFAand
acylglycerolassourcematerialsofbiofuelsandchemi-
cals.12,13)Todate,overexpressionofafattyacidsyn-
thase(FAS)geneinvolvedinthedenovofattyacid
biosynthesisenhancedtheFFAproductivityat2.1-
fold.12)Disruptionofanacyl-CoAsynthetasegene
faaAalsodrasticallyincreasedtheFFAproductivityat
9.2-fold.13)However,constructionofadoublemutant
havingboththeFASoverexpressionandthefaaAdis-
ruptionhasbeendifficultforunknownreasonandbeen
underconstruction.
Thepentosephosphatepathway(PPP)existscom-
monlyinabroadrangeoforganismsandgenerates
bothNADPHandpentosemolecules.Itbeginswith
theglucose6-phosphatemoleculesynthesizedatthe
firstreactionstepofglycolysis,thusPPPcompetesthe
usewiththeglycolysis.PPPconsistsofsevenreaction
stepsuntilitfinallymergesintoglycolysisagain
theproductionofbiodieselfuelandvarious
chemicalssuchaspharmaceuticalsanddietary
supplements.Previously,weattaineda9.2-fold
increaseinfreefattyacidproductivitybydisrupting
apredictedacyl-CoAsynthetasegene(faaA,
AO090011000642)inAspergillusoryzae.Inthis
study,weachievedfurtherincreaseintheproductiv-
itybyoverexpressingapredictedtransketolasegene
ofthepentosephosphatepathwayinthefaaAdis-
ruptant.TheA.oryzaegenomeispredictedtohave
threetransketolasegenesandoverexpressionof
AO090023000345,oneofthethreegenes,resultedin
phenotypicchangeandfurtherincrease(corre-
spondingtoanincreasedproductionof0.38mmol/g
drycellweight)infreefattyacidsat1.4-fold
comparedtothefaaAdisruptant.Additionally,the
biomassofhyphaeincreasedat1.2-foldbytheover-
expression.Asaresult,freefattyacidproduction
yieldperliterofliquidcultureincreasedat1.7-fold
bytheoverexpression.
Keywords:transketolasegeneoverexpression;
pentosephosphatepathway;freefatty
acid;productivity;Aspergillusoryzae
faaAdisruptant
Involvedintheincreasingglobaldemandoffossil
fuels,biofuelsgeneratedfromplantbiomasshavesig-
nificantlybeenfocusedduetolesseffectonglobal
environmentinthecarbonneutralcharacteristics.1,2)In
addition,somepharmaceuticalsanddietarysupple-
mentsproducedfromrarenaturalresourcesareexpen-
sive,andtheirproductionbytheheterologous
microbialbiosynthesisforloweringthepriceshasbeen
desired.3,4)
Microbiallipidproductionhasbeenresearchedin
cerevisiae,8−10)Yarrowialipolytica,11)etc.),filamentous
fattyacid(FFA),acylglycerol,fattyacidmethylester,
andalkanehavebeenthemajortargetsofthemicrobial
production.1)Amongthem,FFAcanbeusedassource
materialsofbiodieselfuelsandchemicals.1)
ThefilamentousfungusA.oryzaeiswellknownfor
highproductioncapabilityofhydrolasessuchasamy-
lase,protease,andlipase.17)Inaddition,A.oryzaeis
notonlyaproducerofsmallmoleculessuchasthe
kojicacid,18)butalsolistedasasafemicroorganism
fortheGRAS(i.e.GenerallyRecognizedasSafe)sta-
tusbytheUSFoodandDrugAdministration.19)Thus,
itimpliesA.oryzaeissuperiortotheotherfilamentous
fungiintheproductionofawiderangeofbiomole-
culesforindustrialapplication.Infact,ithasbeenused
forproducingsake(ricewine),shoyu(soysauce),and
miso(soybeanpaste)byconvertingstarch,proteins,
andlipidsderivedfromrice,wheat,andsoybean,to
manyvarietiesofsmallmoleculesthroughthecatabo-
lismandtheassimilation.
Inourresearch,A.oryzaehasbeenusedforaccom-
plishinghighproductionoflipidssuchasFFAand
acylglycerolassourcematerialsofbiofuelsandchemi-
cals.12,13)Todate,overexpressionofafattyacidsyn-
thase(FAS)geneinvolvedinthedenovofattyacid
biosynthesisenhancedtheFFAproductivityat2.1-
fold.12)Disruptionofanacyl-CoAsynthetasegene
faaAalsodrasticallyincreasedtheFFAproductivityat
9.2-fold.13)However,constructionofadoublemutant
havingboththeFASoverexpressionandthefaaAdis-
ruptionhasbeendifficultforunknownreasonandbeen
underconstruction.
Thepentosephosphatepathway(PPP)existscom-
monlyinabroadrangeoforganismsandgenerates
bothNADPHandpentosemolecules.Itbeginswith
theglucose6-phosphatemoleculesynthesizedatthe
firstreactionstepofglycolysis,thusPPPcompetesthe
usewiththeglycolysis.PPPconsistsofsevenreaction
stepsuntilitfinallymergesintoglycolysisagain
0/5000
Freefattyacidsareusefulassourcematerialsfortheproductionofbiodieselfuelandvariouschemicalssuchaspharmaceuticalsanddietary补充剂。以前，weattaineda9.2 折increaseinfreefattyacidproductivitybydisruptingapredictedacyl-CoAsynthetasegene(faaA,AO090011000642)inAspergillusoryzae.Inthis研究中，weachievedfurtherincreaseintheproductiv-itybyoverexpressingapredictedtransketolasegeneofthepentosephosphatepathwayinthefaaAdis-ruptant。TheA.oryzaegenomeispredictedtohavethreetransketolasegenesandoverexpressionofAO090023000345，oneofthethreegenes，resultedinphenotypicchangeandfurtherincrease(corre-spondingtoanincreasedproductionof0.38mmol/gdrycellweight) infreefattyacidsat1.4 折comparedtothefaaAdisruptant.Additionally,thebiomassofhyphaeincreasedat1.2-foldbytheover-表达式。因此 freefattyacidproductionyieldperliterofliquidcultureincreasedat1.7 折bytheoverexpression。关键词︰ transketolasegeneoverexpression;pentosephosphatepathway; freefatty酸; 生产力;AspergillusoryzaefaaAdisruptantInvolvedintheincreasingglobaldemandoffossil燃料，biofuelsgeneratedfromplantbiomasshavesig.nificantlybeenfocusedduetolesseffectonglobalenvironmentinthecarbonneutralcharacteristics.1,2)In另外，somepharmaceuticalsanddietarysupple-mentsproducedfromrarenaturalresourcesareexpen-平整、 andtheirproductionbytheheterologousmicrobialbiosynthesisforloweringthepriceshasbeendesired.3,4)Microbiallipidproductionhasbeenresearchedin酵母，8−10） Yarrowialipolytica，11) 等)、 丝状指出 (FFA)，甘，fattyacidmethylester，andalkanehavebeenthemajortargetsofthemicrobialproduction.1)Amongthem,FFAcanbeusedassourcematerialsofbiodieselfuelsandchemicals.1)ThefilamentousfungusA.oryzaeiswellknownforhighproductioncapabilityofhydrolasessuchasamy-lase,protease,andlipase.17)Inaddition,A.oryzaeisnotonlyaproducerofsmallmoleculessuchasthekojicacid，18) butalsolistedasasafemicroorganismfortheGRAS (i.e.GenerallyRecognizedasSafe) sta-tusbytheUSFoodandDrugAdministration.19)Thus，itimpliesA.oryzaeissuperiortotheotherfilamentousfungiintheproductionofawiderangeofbiomole-culesforindustrialapplication。事实上 ithasbeenusedforproducingsake （米酒），酱油 （酱油），和味噌 (soybeanpaste) byconvertingstarch，蛋白质，andlipidsderivedfromrice，小麦，andsoybean，到manyvarietiesofsmallmoleculesthroughthecatabo-lismandtheassimilation。Inourresearch,A.oryzaehasbeenusedforaccom-plishinghighproductionoflipidssuchasFFAandacylglycerolassourcematerialsofbiofuelsandchemi-cals.12,13)Todate,overexpressionofafattyacidsyn-这些 (FAS) geneinvolvedinthedenovofattyacidbiosynthesisenhancedtheFFAproductivityat2.1-fold.12) Disruptionofanacyl CoAsynthetasegenefaaAalsodrasticallyincreasedtheFFAproductivityat9.2 fold.13) 然而，constructionofadoublemutanthavingboththeFASoverexpressionandthefaaAdis-ruptionhasbeendifficultforunknownreasonandbeen建设。Thepentosephosphatepathway (PPP) existscom-monlyinabroadrangeoforganismsandgeneratesbothNADPHandpentosemolecules.Itbeginswiththeglucose6-phosphatemoleculesynthesizedatthefirstreactionstepofglycolysis thusPPPcompetestheusewiththeglycolysis。PPPconsistsofsevenreactionstepsuntilitfinallymergesintoglycolysisagain
正在翻譯中..
Freefattyacidsareusefulassourcematerialsfor
theproductionofbiodieselfuelandvarious
chemicalssuchaspharmaceuticalsanddietary
supplements.Previously，weattaineda9.2倍
increaseinfreefattyacidproductivitybydisrupting
apredictedacyl-CoAsynthetasegene（FAAA，
AO090011000642）inAspergillusoryzae.Inthis
研究，weachievedfurtherincreaseintheproductiv-
itybyoverexpressingapredictedtransketolasegene
ofthepentosephosphatepathwayinthefaaAdis-
ruptant.TheA.oryzaegenomeispredictedtohave
threetransketolasegenesandoverexpressionof
AO090023000345，oneofthethreegenes，resultedin
phenotypicchangeandfurtherincrease（任意一台
spondingtoanincreasedproductionof0.38mmol /摹
drycellweight）infreefattyacidsat1.4倍
comparedtothefaaAdisruptant.Additionally的
biomassofhyphaeincreasedat1.2-foldbytheover-
expression.Asaresult，freefattyacidproduction
yieldperliterofliquidcultureincreasedat1.7倍
bytheoverexpression。
关键词：transketolasegeneoverexpression;
pentosephosphatepathway; freefatty
酸;生产力;米曲霉
faaAdisruptant
Involvedintheincreasingglobaldemandoffossil
燃料，biofuelsgeneratedfromplantbiomasshavesig-
nificantlybeenfocusedduetolesseffectonglobal
environmentinthecarbonneutralcharacteristics.1,2）在
此外，somepharmaceuticalsanddietarysupple-
mentsproducedfromrarenaturalresourcesareexpen-
西伯，andtheirproductionbytheheterologous
microbialbiosynthesisforloweringthepriceshasbeen
desired.3,4）
Microbiallipidproductionhasbeenresearchedin
酵母，8-10）Yarrowialipolytica，11）等），丝状
fattyacid（FFA），酰基甘油，fattyacidmethylester，
andalkanehavebeenthemajortargetsofthemicrobial
生产0.1）Amongthem，FFAcanbeusedassource
materialsofbiodieselfuelsandchemicals.1）
ThefilamentousfungusA.oryzaeiswellknownfor
highproductioncapabilityofhydrolasessuchasamy-
化酶，蛋白酶，andlipase.17）Inaddition，A.oryzaeis
notonlyaproducerofsmallmoleculessuchasthe
曲酸，18）butalsolistedasasafemicroorganism
fortheGRAS（ieGenerallyRecognizedasSafe）台站
tusbytheUSFoodandDrugAdministration.19）因此，
itimpliesA.oryzaeissuperiortotheotherfilamentous
fungiintheproductionofawiderangeofbiomole -
culesforindustrialapplication.Infact，ithasbeenused
forproducingsake（ricewine），酱油（酱油）和
MISO（soybeanpaste）byconvertingstarch，蛋白质，
andlipidsderivedfromrice，小麦，andsoybean，以
manyvarietiesofsmallmoleculesthroughthecatabo-
。lismandtheassimilation
Inourresearch，A.oryzaehasbeenusedforaccom-
plishinghighproductionoflipidssuchasFFAand
acylglycerolassourcematerialsofbiofuelsandchemi-
cals.12,13 ）TODATE，overexpressionofafattyacidsyn-
合酶（FAS）geneinvolvedinthedenovofattyacid
biosynthesisenhancedtheFFAproductivityat2.1-
fold.12）Disruptionofanacyl-CoAsynthetasegene
faaAalsodrasticallyincreasedtheFFAproductivityat
9.2 fold.13）然而，constructionofadoublemutant
havingboththeFASoverexpressionandthefaaAdis-
ruptionhasbeendifficultforunknownreasonandbeen
underconstruction。
Thepentosephosphatepathway（PPP）existscom-
monlyinabroadrangeoforganismsandgenerates
bothNADPHandpentosemolecules.Itbeginswith
theglucose6，phosphatemoleculesynthesizedatthe
firstreactionstepofglycolysis ，thusPPPcompetesthe
usewiththeglycolysis.PPPconsistsofsevenreaction
stepsuntilitfinallymergesintoglycolysisagain
theproductionofbiodieselfuelandvarious
chemicalssuchaspharmaceuticalsanddietary
supplements.Previously，weattaineda9.2倍
increaseinfreefattyacidproductivitybydisrupting
apredictedacyl-CoAsynthetasegene（FAAA，
AO090011000642）inAspergillusoryzae.Inthis
研究，weachievedfurtherincreaseintheproductiv-
itybyoverexpressingapredictedtransketolasegene
ofthepentosephosphatepathwayinthefaaAdis-
ruptant.TheA.oryzaegenomeispredictedtohave
threetransketolasegenesandoverexpressionof
AO090023000345，oneofthethreegenes，resultedin
phenotypicchangeandfurtherincrease（任意一台
spondingtoanincreasedproductionof0.38mmol /摹
drycellweight）infreefattyacidsat1.4倍
comparedtothefaaAdisruptant.Additionally的
biomassofhyphaeincreasedat1.2-foldbytheover-
expression.Asaresult，freefattyacidproduction
yieldperliterofliquidcultureincreasedat1.7倍
bytheoverexpression。
关键词：transketolasegeneoverexpression;
pentosephosphatepathway; freefatty
酸;生产力;米曲霉
faaAdisruptant
Involvedintheincreasingglobaldemandoffossil
燃料，biofuelsgeneratedfromplantbiomasshavesig-
nificantlybeenfocusedduetolesseffectonglobal
environmentinthecarbonneutralcharacteristics.1,2）在
此外，somepharmaceuticalsanddietarysupple-
mentsproducedfromrarenaturalresourcesareexpen-
西伯，andtheirproductionbytheheterologous
microbialbiosynthesisforloweringthepriceshasbeen
desired.3,4）
Microbiallipidproductionhasbeenresearchedin
酵母，8-10）Yarrowialipolytica，11）等），丝状
fattyacid（FFA），酰基甘油，fattyacidmethylester，
andalkanehavebeenthemajortargetsofthemicrobial
生产0.1）Amongthem，FFAcanbeusedassource
materialsofbiodieselfuelsandchemicals.1）
ThefilamentousfungusA.oryzaeiswellknownfor
highproductioncapabilityofhydrolasessuchasamy-
化酶，蛋白酶，andlipase.17）Inaddition，A.oryzaeis
notonlyaproducerofsmallmoleculessuchasthe
曲酸，18）butalsolistedasasafemicroorganism
fortheGRAS（ieGenerallyRecognizedasSafe）台站
tusbytheUSFoodandDrugAdministration.19）因此，
itimpliesA.oryzaeissuperiortotheotherfilamentous
fungiintheproductionofawiderangeofbiomole -
culesforindustrialapplication.Infact，ithasbeenused
forproducingsake（ricewine），酱油（酱油）和
MISO（soybeanpaste）byconvertingstarch，蛋白质，
andlipidsderivedfromrice，小麦，andsoybean，以
manyvarietiesofsmallmoleculesthroughthecatabo-
。lismandtheassimilation
Inourresearch，A.oryzaehasbeenusedforaccom-
plishinghighproductionoflipidssuchasFFAand
acylglycerolassourcematerialsofbiofuelsandchemi-
cals.12,13 ）TODATE，overexpressionofafattyacidsyn-
合酶（FAS）geneinvolvedinthedenovofattyacid
biosynthesisenhancedtheFFAproductivityat2.1-
fold.12）Disruptionofanacyl-CoAsynthetasegene
faaAalsodrasticallyincreasedtheFFAproductivityat
9.2 fold.13）然而，constructionofadoublemutant
havingboththeFASoverexpressionandthefaaAdis-
ruptionhasbeendifficultforunknownreasonandbeen
underconstruction。
Thepentosephosphatepathway（PPP）existscom-
monlyinabroadrangeoforganismsandgenerates
bothNADPHandpentosemolecules.Itbeginswith
theglucose6，phosphatemoleculesynthesizedatthe
firstreactionstepofglycolysis ，thusPPPcompetesthe
usewiththeglycolysis.PPPconsistsofsevenreaction
stepsuntilitfinallymergesintoglycolysisagain
正在翻譯中..
freefattyacidsareusefulassourcematerialsfortheproductionofbiodieselfuelandvariouschemicalssuchaspharmaceuticalsanddietary此前，weattaineda9.2-fold补充。increaseinfreefattyacidproductivitybydisruptingapredictedacyl CoAsynthetasegene（岛，ao090011000642）inaspergillusoryzae.inthis研究weachievedfurtherincreaseintheproductiv—itybyoverexpressingapredictedtransketolasegeneofthepentosephosphatepathwayinthefaaadis—ruptant.thea.oryzaegenomeispredictedtohavethreetransketolasegenesandoverexpressionofao090023000345，oneofthethreegenes，导致phenotypicchangeandfurtherincrease（相关—spondingtoanincreasedproductionof0.38mmol/gdrycellweight）infreefattyacidsat1.4-foldcomparedtothefaaadisruptant。此外，该biomassofhyphaeincreasedat1.2-foldbytheover—Asaresult，freefattyacidproduction表达。yieldperliterofliquidcultureincreasedat1.7-foldbytheoverexpression。关键词：transketolasegeneoverexpression；磷酸戊糖路径；脂肪酸性条件；生产力；faaadisruptantinvolvedintheincreasingglobaldemandoffossil燃料，biofuelsgeneratedfromplantbiomasshavesig—nificantlybeenfocusedduetolesseffectonglobalenvironmentinthecarbonneutralcharacteristics 1,2）中。此外，somepharmaceuticalsanddietarysupple—mentsproducedfromrarenaturalresourcesareexpen—性，andtheirproductionbytheheterologousmicrobialbiosynthesisforloweringthepriceshasbeen需4）。microbiallipidproductionhasbeenresearchedin酵母，8−10）yarrowialipolytica，11）等），丝状脂肪酸（FFA）、甘油脂、fattyacidmethylester，andalkanehavebeenthemajortargetsofthemicrobial生产。1）其中，FFAcanbeusedassourcematerialsofbiodieselfuelsandchemicals 1）。thefilamentousfungusa.oryzaeiswellknownforhighproductioncapabilityofhydrolasessuchasamy—酶，蛋白酶，和17）此外，A.oryzaeis。notonlyaproducerofsmallmoleculessuchasthe曲酸，18）butalsolistedasasafemicroorganismforthegras（i.e.generallyrecognizedassafe）站—19）因此tusbytheusfoodanddrugadministration，itimpliesa.oryzaeissuperiortotheotherfilamentousfungiintheproductionofawiderangeofbiomole—culesforindustrialapplication。事实上，ithasbeenusedforproducingsake（黄酒），酱油（酱油），和味噌（soybeanpaste）byconvertingstarch，蛋白质，andlipidsderivedfromrice，小麦，大豆，以manyvarietiesofsmallmoleculesthroughthecatabo—lismandtheassimilation。inourresearch，a.oryzaehasbeenusedforaccom—plishinghighproductionoflipidssuchasffaandacylglycerolassourcematerialsofbiofuelsandchemi—迄今为止，overexpressionofafattyacidsyn CALS 12,13）—酶（FAS）geneinvolvedinthedenovofattyacidbiosynthesisenhancedtheffaproductivityat2.1—倍。12）Disruptionofanacyl CoAsynthetasegenefaaaalsodrasticallyincreasedtheffaproductivityat13）然而，constructionofadoublemutant 9.2倍。havingboththefasoverexpressionandthefaaadis—ruptionhasbeendifficultforunknownreasonandbeen正在建设中的。thepentosephosphatepathway（PPP）existscom—monlyinabroadrangeoforganismsandgeneratesbothnadphandpentosemolecules。itbeginswiththeglucose6 phosphatemoleculesynthesizedatthefirstreactionstepofglycolysis，thuspppcompetestheusewiththeglycolysis.pppconsistsofsevenreactionstepsuntilitfinallymergesintoglycolysisagain
正在翻譯中..
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