不同氮源对异养小球藻生物量和油脂积累的影响

366Vol.36,No.6201211ACTAHYDROBIOLOGICASINICANov.,2012收稿日期:2011-08-02;修订日期:2012-07-09基金项目:(08JCYBJC10600)作者简介:(1986),,;;E-mail:[e-mail]通讯作者:,,;E-mail:[e-mail]:10.3724/SP.J.1035.2012.01027朱义平宋东辉杨国兰(,,300457):,,,,NaNO3,SENaNO31.5mmol/L3.0mmol/L6.0mmol/L,0.75mmol/L1.5mmol/L3.0mmol/L10d::(1),,NaNO31.5mmol/L,2.65g/L51.21%,1.36g/L(2),,,(3),,38.78%47.02%,:;;;;中图分类号:Q142文献标识码:A文章编号:1000-3207(2012)06-1027-08,,,,50%,80%[1],,,,Lu,etal.[2],,,Shen,etal.[3],2.4g/LKNO3MB654mg/L/dXiong,etal.[4],4g/L8.28g/L,,,,,,10283611.1(Chlorellavul-garisFACHB-7),SE[5]:0.75g/LNaNO3,0.23g/LMgSO4·7H2O,0.23g/LK2HPO4,0.53g/LKH2PO4,0.03g/LNaCl,0.01g/LFeSO4·6H2O,2.0mL/LFe-EDTA,1.0mL/LA5solution[2]:20.0g4,pH6.07.0,60℃,,,20min,0.1%0.2%α-,30min,20min,HClpH4.55.0,0.4%,,60,℃1h4000r/min5min,,1.2SE,1.53.06.0mmol/LNaNO3;NaNO3,1.53.06.0mmol/L0.751.53.0mmol/L,pH6.0,120℃20min10%(1.6×106cell/mL)100mL,(20g/L)(25±1)℃150r/min,10d1.3[6]:μ=(lnNt2−lnNt1)/t:μ,;Nt2,t2(cell/L);Nt1,t1(cell/L)t,10d(240h),t=101.448h3.0mL(9.0×106/mL),4000r/min10min,3.0mL,4℃,752,665nm652nmOD,Wellburn,etal.[7]:C(a+b)(mg/L)=1.44A665+24.93A6521.5(9-diethylamino-SH-benzo[a]penoxazine-5-one)[810]:5.0mL(9.0×106/mL),250mLDMSO,500μL(0.1mg/mL),60℃1min,480nm580nm,580nm1.6-[11]0.3g,3.0mL(12),40℃,,10%,,4000r/min10min,,60℃,,:=(/)×100%22.1NaNO3,(1)6.0mmol/LNaNO3,240h,;NaNO33.0mmol/L,240h,,,,,,;(1.56.0mmol/LNaNO3),,,,(1),ab,,(2)3.0mmol/L6.0mmol/LNaNO3,6:1029,6mmol/L(21.46pg/cell),(1),(11),NaNO3144h,(3)6.0mmol/L,,,(1),,1NaNO3Fig.1GrowthcurvesofC.vulgarisunderdifferentNaNO3con-centrations2NaNO3Fig.2TotalchlorophyllcontentsofC.vulgarisunderdifferentNaNO3concentrations(4),,144h,(3),(4),,3Fig.3GrowthcurvesofC.vulgarisunderdifferentalaninecon-centrations4Fig.4TotalchlorophyllcontentsofC.vulgarisunderdifferentalanineconcentrations,NaNO3,,0.75mmol/L144h,3.0mmol/L(5)5Fig.5GrowthcurvesofC.vulgarisunderdifferenttyrosineconcentrations103036表1不同条件下小球藻比生长速率和单位细胞叶绿素含量变化Tab.1GrowthratesandchlorophyllcontentsofC.vulgarisunderdifferentnutrientlevelsNitrogensources1.5mmol/LNaNO33.0mmol/LNaNO36.0mmol/LNaNO31.5mmol/Lalanine3.0mmol/Lalanine6.0mmol/Lalanine0.75mmol/Ltyrosine1.5mmol/Ltyrosine3.0mmol/LtyrosineSpecialgrowthrates0.740.850.860.490.520.590.460.400.34Chlorophyllcontent,pg/cell15.1016.6821.4613.2116.8816.7212.3315.6229.89,,1.5mmol/L,2.57mg/L;3.0mmol/L,,4.28mg/L(6),3.0mmol/L,12.33pg/cell0.75mmol/L(1)6Fig.6TotalchlorophyllcontentsofC.vulgarisunderdifferenttyrosineconcentrations,(1),,0.740.86,,,,,,,,,,,2.2,,NaNO3(7)NaNO31.5mmol/L,2.65g/L-,,51.21%,1.36g/L(2),-,9.55×103a.u.7.07a.u.(7),NaNO36.0mmol/L,4.94g/L,13.65%,(2)NaNO3,,,,,,0.100.14g/L,,NaNO3710(2)1.5mmol/L,2.73a.u.(30.78%),3.0mmol/L6.0mmol/L,0.46g/L0.50g/L,1.25a.u.0.10a.u.,28.72%19.50%(27),,,,,1.5mmol/L6.0mmol/L,0.23g/L,2.422.86a.u.,3.0mmol/LNaNO3(2.82a.u.)1.5mmol/L(2.73a.u.)(7),38.78%47.02%(2),6:1031表2不同氮源对小球藻生物量和油脂含量的影响Tab.2EffectsofdifferentnitrogensourcesonbiomassaccumulationandlipidcontentofC.vulgarisNitrogensourceDrycellweight(g/L)Lipidcontent(%)Lipidweight(g/L)1.5mmol/LNaNO32.6551.211.363.0mmol/LNaNO34.5627.671.266.0mmol/LNaNO34.9413.650.671.5mmol/L0.4430.780.143.0mmol/L0.4628.720.146.0mmol/L0.5019.500.100.75mmol/L0.2547.020.121.5mmol/L0.2346.420.113.0mmol/L0.2338.780.097Fig.7EffectsofdifferentnitrogensourcesonbiomassaccumulationandfluorescenceintensityofC.vulgariscells0.230.25g/L,,,,(8),,,[12]3,,,30%60%[13,14]8(×400)Fig.8OpticalmicroscopephotographsofC.vulgarisunderdif-ferentnitrogensources(×400)(1)NaNO3(2)(3)(1)NaNO3asthenitrogensource(2)alanineasthenitrogensource(3)tyrosineasthenitrogensource103236,,,:NaNO31.5mmol/L,2.65g/L,51.21%(2),(C.protothecoides)(C.vulgaris),,,,[17][15,16][15][9],,,,Liu,etal.[9]25d56.6%,,10d,Qiao,etal.[18]23.7%,Liang,etal.[19]2g/L,54mg/L/d,2.65g/L,51.21%,Lu,etal.[12](Chlorellaprotothecoides),(4.26±0.18)g/L(2.14±0.11)g/L,2.65g/L1.36g/L,,,,,,,Greenspan,etal.[20],[21][22][9,20],,,,[2325],,,,,,,,[2],,,[26],,10,,,[26],,,,,,:[1]LiXF,XuH,WuQY.Large-scalebiodieselproductionfrommicroalgaChlorellaprotothecoidesthroughheterotro-phiccultivationinbioreactors[J].BiotechnologyandBioen-gineering,2007,98(4):764771[2]LuY,ZhaiY,LiuMS,etal.Biodieselproductionfromalgaloilusingcassava(ManihotesculentaCrantz)asfeedstock[J].JournalofAppliedPhycology,2010,22(5):573578[3]ShenY,YuanW,PeiZ.HeterotrophiccultureofChlorellaprotothecoidesinvariousnitrogensourcesforlipidproduc-tion[J].AppliedBiochemistryandBiotechnology,2010,160(6):1674684[4]XiongW,LiXF,XiangJY,etal.High-densityfermentationofmicroalgaChlorellaprotothecoidesinbioreactorformi-crobio-dieselproduction[J].AppliedMicrobiologyandBio-technology,2008,78(1):2936[5]HuangK.Studyonmethodsandmechanismforremovalnitrogenandphosphorusinwastewaterbyalgae.ThesisforMasterofScience[D].NanchangUniversity,Nanchang.2007[..6:1033,,.2007][6]YeLC,YeJA,XuGZ,etal.EffectsofnitrogensourcesongrowthofalgaChlorellavulgaris[J].FisheriesScience,2007,26(6):319322[,,,..,2007,26(6):319—322][7]AlanRW.Thespectraldeterminationofchlrorophyllsaandb,aswellastotalcarotenoids,usingvarioussolventswithspectrophotometersofd