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392Vol.39,No.220153ACTAHYDROBIOLOGICASINICAMar.,2015收稿日期:2014-04-30;修订日期:2014-08-23基金项目:(2011CB200901)作者简介:(1982),;;;E-mail:lifj_hit@163.com通信作者:,E-mail:hanhuahu@ihb.ac.cndoi:10.7541/2015.58CO2李凤娟1万秀2方仙桃2胡锐2高大文1胡晗华2(1.,150090;2.,430072)EFFECTSOFNITROGENSOURCESANDCO2LEVELSONTHEGROWTHANDOILACCUMULATIONINNANNOCHLOROPSISLIMNETICALIFeng-Juan1,WANXiu2,FANGXian-Tao2,HURui2,GAODa-Wen1andHUHan-Hua2(1.StateKeyLaboratoryofUrbanWaterResourceandEnvironment,HarbinInstituteofTechnology,Harbin150090,China;2.InstituteofHydrobiology,ChineseAcademyofSciences,Wuhan430072,China):;;;Keywords:Nannochloropsislimnetica;Triacylglycerols;Biodiesel;NitrogenUtilization中图分类号:Q948.1文献标识码:A文章编号:1000-3207(2015)02-0436-05CO2,SOxNOx[1]NOx[2],,,,,[3],,(Nannochloropsis)(Eustigmatophyceae),[4],,,[5—8],[4]6[9],[10],,,(N.limnetica),,CO2,,11.1(N.limneticaKR1998/3)(InstituteofFreshwaterEcologyandInlandFisheries,Neuglobsow)LotharKrienitz,BG11,22℃60μmolphotons/(m2·s)1mmol/LNH4ClNaNO2NaNO3BG11NaNO33cm,60cmA730=0.15,2:CO2437300mL2%5%CO2(v/v),120μmolphotons/(m2·s)pH1.2A730pHpHNO2−NO3−(A220)[11]NH4+-(A420)[12][13]1.32468,,BlighDyer[14]6000r/min︰(1︰1),1—2min,3/101mol/L0.2mol/L,12000r/min5min,EP,,–20℃[15]Silicagel60F254(MerkKgaADarmstadt,Germany),︰︰(70︰30︰1,v/v/v),,37℃5—10min[16](Glyceryltrioleate)Sigma1.46000r/min8d,ddH2O,,[14]100mg:(1︰2),ddH2O,,,N2,,:1.5mLAglient,1mL1mol/L,N2,100℃1h,200μL,,200μL,,N2100μL,1μLUltraTrace(ThermoScientific,UnitedStates)(60m×0.25mm);250,FID℃280;℃:150,1min;150℃—220,5/min;220℃℃—280,℃20/min;280,3℃℃3min;,2mL/min22.1,CO25%,,55%CO2,[17],CO2()[18],2%CO2(0—0.28/d)(0.27—0.29/d)NH4Cl,2%CO2,pH6(1),CO2,CO2[19]pH7—8.5,2%CO2,pH6—7.5NH4ClpH,1NH4ClpHNaNO2NaNO311(A)2%CO2(B)(A730,)pH()Fig.1Growth(A730)andpHvalueinmediumofNannochloropsislimneticaculturedatdifferentnitrogensourcesaeratedwithair(A)and2%CO2(B)43839NaNO2,,,[20]NO3−NO2−,NH4+,NH4+,[21,22],[23],1mmol/LNaNO2,NOx[24]2.22,,,,,,4,[22],,,(2)2(A)2%CO2(B)()()Fig.2NitrogenandphosphateconcentrationsinmediumofNannochloropsislimneticaculturedatdifferentnitrogensourcesaeratedwithair(A)and2%CO2(B)2.3,,13(DGAT),,[25,26]28,4,(3)4,,200μmol/L2,,[27],17.6mmol/L,[28],2%CO2,,3(A)2%CO2(B)Fig.3ComparisonoflipidcompositionandcontentinNan-nochloropsislimneticaculturedatdifferentnitrogensourcesaer-atedwithair(A)and2%CO2(B)M.Triolein2:CO2439表1不同氮源下湖泊微拟球藻在通空气(A)与2%CO2(B)条件下总脂含量(占藻细胞干重)及其脂肪酸组成(摩尔百分比)Tab.1Totallipid(%drycellweight)contentandfattyacidcomposition(mol%)intotallipidsofNannochloropsislimneticaKR1998/3culturedatdifferentnitrogensourcesaeratedwithairor2%CO2NO2−NO3−NH4+TotallipidsandfattyacidsAir2%CO2Air2%CO2AirC14:06.44±1.729.12±0.352.78±3.338.79±0.338.50±4.48C14:10.59±0.400.35±0.380.49±0.590.26±0.220.40±0.38C16:029.56±2.3226.56±1.4126.62±2.6431.01±0.9028.72±2.86C16:132.90±1.4031.78±3.0532.80±2.4531.30±0.5934.86±3.78C18:01.28±0.411.72±1.211.87±0.791.50±0.031.10±0.76C18:114.44±0.4812.88±1.2216.18±1.7312.21±1.5813.48±2.89C18:22.46±0.492.84±0.952.99±1.392.55±0.342.67±0.37γ-C18:30.11±0.120.25±0.280.97±1.320.07±0.021.06±0.89α-C18:30.71±0.171.28±0.951.25±1.270.69±0.020.69±0.66C18:40.44±0.520.33±0.460.96±1.060.20±0.280.36±0.51C20:01.05±1.030.71±0.660.86±0.870.65±0.070.57±0.64C20:42.73±0.883.46±0.143.51±1.162.95±0.032.21±1.64C20:56.79±2.297.70±0.127.25±0.097.52±0.984.84±3.41C22:00.09±0.130.32±0.460.28±0.380.00±0.000.23±0.33C22:40.10±0.140.26±0.370.37±0.230.00±0.000.05±0.08C22:50.24±0.340.37±0.200.55±0.630.28±0.080.19±0.27C22:60.08±0.100.07±0.070.28±0.380.03±0.010.05±0.06(%drycellweight)40.339.144.834.141.9,8d134%—45%,[27]CO2C16:0C16:1C18:1,71%—77%C14:0C20:5(EPA),EPA[29]EPA5%—8%,,EPA[30],12%,,,,,:[1]KoornneefJ,RamirezA,vanHarmelenT,etal.TheimpactofCO2captureinthepowerandheatsectorontheemissionofSO2,NOx,particulatematter,volatileorganiccompoundsandNH3intheEuropeanUnion[J].AtmosphericEnviron-ment,2010,44(11):1369—1385[2]JiangR,HuangSB,FanLR.RemovalofNOxfromfluegasbyabiofilterunderaerobicconditions[J].ActaScientiaeCircumstantiae,2007,27(9):1469—1475[,,.NOx.,2007,27(9):1469—1475][3]ChistiY.Biodieselfrommicroalgae[J].BiotechnologyAdvances,2007,25:294—306[4]JinkersonRE,RadakovitsR,PosewitzMC.Genomicin-sightsfromtheoleaginousmodelalgaNannochloropsisgaditana[J].Bioengineered,2013,4:37—43[5]RadakovitsR,JinkersonRE,FuerstenbergSI,etal.Draftgenomesequenceandgenetictransformationoftheoleagi-nousalgaNannochloropsisgaditana[J].NatureCommuni-cations,2012,3:686[6]VielerA,WuG,TsaiCH,etal.Genome,functionalgeneannotation,andnucleartransformationoftheHeterokontoleaginousalgaNannochloropsisoceanicaCCMP1779[J].PLoSGenetics,2012,8:e1003064[7]LiF,GaoD,HuH.High-efficiencynucleartransformationoftheoleaginousmarineNannochloropsisspeciesusingPCRproduct[J].Bioscience,Biotechnology,andBiochemis-try,2014,78(5):812—81744039[8]RodolfiL,ChiniZittelliG,BassiN,etal.Microalgaeforoil:strainselection,inductionoflipidsynthesisandoutdoormasscultivationinalow-costphotobioreactor[J].Biotech-nologyandBioengineering,2009,102:100—112[9]AndersenRA,BrettRW,PotterD,etal.PhylogenyoftheEustigmatophyceaebasedupon18SrDNA,withemphasisonNannochloropsis[J].Protist,199