微生物发酵降解植物甾醇侧链生产17_酮甾体研究进展

8520109ChineseJournalofBioprocessEngineeringVo.l8No.5Sep.2010do:i10.3969/.jissn.1672-3678.2010.05.015:2010-03-03:;(08ZQ026-023);:(1943),,,,:,Emai:[e-mail]17杨顺楷1,杨亚力1,吴中柳1,范林萍2,李小刚2(1.中国科学院成都生物研究所,成都610041;2.兰州大学生命科学学院,兰州730000):微生物发酵降解植物甾醇侧链,生产雄甾4烯3,17二酮(AD),雄甾1,4二烯3,17二酮(ADD),和9羟基AD甾体药物中间体的工业生物技术对改变制造甾体激素药物半合成原料薯蓣皂素短缺的现状,实现甾体激素药物半合成原料多元化,合理利用我国甾体植物资源具有重要意义重点评述了近期微生物法断植物甾醇侧链制ADADD和9羟基AD的研究现状,内容包括:1)微生物菌种选育;2)菌种相关的细胞生理,酶学性质和生物催化过程;3)相关酶的细胞定位及生物反应器;4)发酵工艺选择和甾醇原料的合理利用:生物降解;微生物发酵;植物甾醇:TQ4642:A:1672-3678(2010)05-0069-09Microbialfermentationofphytosterolsidechaincleavedforproductionof17ketosteroidsYANGShunkai1,YANGYali1,WUZhongliu1,FANLinping2,LIXiaogang2(1.ChengduInstituteofBiology,ChineseAcademyofSciences,Chengdu610041,China;2.SchoolofLifeSciences,LanzhouUniversity,Lanzhou730000,China)Abstract:Thestudyonthemicrobialdegrationofphytosterolsidechainandthedevelopmentofindustralbiotechnologyforproductionof17ketosteroids,.ie.,androst4ene3,17dione(AD),androst1,4diene3,17dione,and9hydroxyAD,areofsignificancetoreplacepartiallydiosgeninasitsshortagewiththem,andtoperformmultiplesourcesasonekindofrawmateralsofsemisyntheticproductioninsteroidpharmaceuticalindustry,andtoputplantsteroidresourcestorationaluseinChinafortoday.Thispaperreviewedmicrobialorganismforthebreedingandselectionmutan,tthestrainselectionofrelatedcytologicalphysiology,enzymepropertiesandbiocatalyticalprocessing,relatedenzymelocationincellandbioreacterselection,andfermentationalprocessselectionandrationaluseofsterolrawmaterials.Keywords:biodegration;fermention;phytosterol,,,43,17(AD),1,43,17(ADD)9AD,,,(1)[1-2],1Fig.1TimecourseofcellulaseproductionbyT.reesei,AD(D),,AD(D),9OHAD,FCl,9OH,C9,11,C9,C11C11,9(FCl),C11,/[3-5]1979,9OHAD,7,,367%[6]117Table1Fermentationproductionof17ketosteroids/(gL-1)/%7,9(11)3025NRRLB38054,8(14)ADD3,1730319505Moruxellasp1510NRRLB38055103NRRLB3805AD3503NRRLB3683ADD3010NRRLB3805AD2510NRRLB3683ADD2010NRRLB3805AD9050Mycobacteriumsp.VKMAc1815DET1AD7210NRRLMB3683AD9030NRRLMB3683AD80,,,,,[7],,,,[5],C17(AD(D),9OHAD)708,,40,2080ADADD,1000t/a,21,(),ADADD,200m3[8],50m3AD,11OHAD,C17,[9],9OHAD1!∀,NRRL-3805,(UV),1(2)9M.vaccaeZIMET11052,110539OHAD,9OHAD1(2)M.vaccae,,9OHAD,[10]II,(CICC)(Mycobacteriumfortuitum)CICC10279,,(),9OHAD,TLC,,9OHAD,9OHAD(IR,1HNMR,13CNMR),[11]9OHAD,()Donova[12]9OHAD,,,9OHADsp.Ac1815D:6/26/210,S9OHAD(6/2S)6/2SUV,,,6/21UV9OHAD27%9OHAD3d,#596%9OHAD,6/21UV,,SAD/9OHAD,,,sp.24MVS,9OHADsp.Ac1815DAD,9OHAD,91(2),1815DNaI50RGm10R()Rif100RGm10R(),AD9OHADC,,9OHAD9OHAD,,,(EMS)CUV,,AD/9OHAD,,24M,9OHADsp.,715:17,,9OHAD,99OHAD,,UV,(50%)1815D24MAD9OHADC,,9OHAD,9OHAD1(2),9OH,24MADD9ADD,1AD,AD99OHAD1Egorova[13]24M,9M.roseum,9[14](R.erythropolis)Stenone,9KshB[15]9OHStenoneC179(10),C179,,9OHADAD9,9OHADC179C1724MAD,(025g/L),AD9OHAD7%,5g/L,9OHAD50%,AD9OHAD24M,9,9,9993,:,9OHAD[12]2,ADADD9,AD(D),,(XAD,AmberliteEP60)AD(D),AD(D)ADAD(Schizosaccharomycespombe),,[12],Egorova[13]AD91(2)AD,9OHADDA,B,9(10),CO2H2O(2)AD91(2)?,231,2SDH1SDH2,AD1,2(AD∃ADD)9OHAD1,2(9OHADD),,91,2,AD[16-18]NRRLB3805VKMAc1815DNRRLB3683AD,,,,,,AD(D)72819hydroxylase;23ketosteroid1,2dehydrogenase;33ketosteroid1,2dehydrogenase;41enesteroidreductase;517hydroxysteroiddehydrogenase2spp.Fig.2SitosteroltransformationbyMycobacteriumspp.,,NRRLB3685AD,(MMS),12,,11(2),AD,AD,ADDADVKMAc1815D,AD,63%~68%,EMSC,AD,70%~75%,ADC17,3,1717AD(D),,1(2),,,VKMAc1817D9OHAD,1(St1DH),9OHAD,AD,(PMS),St1DHPMS9OHADVmax,ADADD,VKMAcSt1DH9OHAD,AD30,,ADD,1(2),,St1DH9OHADAD,Worcha[19]St1DH3St1DH,3,9OH3ATCC6842CO2H2O,HA12735:17St1DH:,9[20]NRRL8153,9OHADSt1DH[21]Ac1817D9OHAD,St1DH9OH3,VKMAc1817D,VKMAc1817D,9OH3St1DH,AD[13],,3,Sukhodolskaya[20]VKMAc1817D9OHADSt1DH,St1DH,St1DH,St1DH,St1DH(NH4)2SO4,DEAE,BioGelA05M,St1DH28,(58%2)&103,(N.coralline)St1DH605&104,(R.erythropolis)56&104,,Ac1817DSt1DH,St1DH,,9OHAD,Angelova[21]AD99,9C9,,,,,(),,9AD9OHAD,CN,9AD,pH,[21],75%,N,85%,,9OHAD,9OHADAD,2:9!191,43,17(9OHADD),9,10,B9OHADDC1AD,9OHADADC1,9;AD9,9OHAD,C19OHADD92[21]44.1,29OHAD,,,(),AD;,()AD9,,9OHAD,,,M.roseum,,9OHAD31(2),7489OHADB8119M.vccae23,171(2),AD9OHADSQ231(2),39[15,22]2IA,KshAKshBKshA(RG2),ADADD,CO2H2O(9)KshB(RG4),94.2Angelova[23]AD,,621!1,Houng[24](A.ochraceus)11,,9Angelova[21],,AD99!1,C/N,!1,9,!1(3∋1),1g/LAD,90%:!1,(60%),,,3%(),9!1,AD,,AD:AD,AD,AD6,AD,,,,,,,,C[23]4.3Donova[25],C17sp.1817D(300~350h),,14%(),,755:17,,9OHAD,10g/L1817D,60h9OHAD65%,C17,[8],,C17,C17,C171000t[25],60%,C17,,,C17,,!∀[5],,,,!∀C17,,C17(ADADD)MB3683,()(96%),70%NRRL3805ADADD,,(ADADD)25mmol/L55%;[5],,,,,AD,ADD,5,UV,9OHAD(),17,ADADD9OHAD,,,,,AD(D),9OHAD:[1],.[J].,2009,17(24):56.YangShunka,iYangYal.iProblemsappearindevelopingsterolsresources[J].FineandSpecialtyChemicals,2009,17(24):56.[2],,,.蔴皂素资源的[J].,2007,19(4):696700.YangYal,iYangWeil,iHuangDeying,eta.lStudyontheresourceofhecogenininJinsaRiverDryhotvalley[J].NaturalProductResearchandDevelopment,2007,19(4):696700.[3]SlijkhuisH,BerkelAE,RodenrijsEN,eta.lPreparationof9alphahydroxy17ketosreroidsusingMycobacteriumspeciesCBS482.86:US,5298398[P].19940329.[4],.[J].,2006,33(2):142146.ZhangYuqing,WangDongqin