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562001,Vol.21,No.1孙艳钱世钧(,100080)本文综述了以苯取代苯联苯和多环芳烃为代表的芳香族化合物的生物降解途径,其共同之处在于经过两步双加氧酶作用,生成二醇和开环两步双加氧酶分别为芳环羟基化双加氧酶和芳环断裂双加氧酶以甲苯途径为代表讨论了芳香族化合物的分子生物学研究情况代谢工程研究是九十年代兴起的芳香族化合物生物降解的研究内容,通过对甲苯途径的代谢工程研究明确了途径中的关键酶,并通过对关键酶的活性提高使整个途径的代谢流增加芳香族化合物生物降解双加氧酶分子生物学代谢工程1,,,:,,,,,1111[1]1(a),,,112:,,422~71(b)[2]C7,XB,[3]1131(c),[4]:1,23,4,,2--6--6--2,3-(HOPDA),(BA),,114(PolycyclicAromaticHydrocarbons,PAH)1(d)[5],1,2-,19223,NADH[2Fe-S],:11,NADHNADPHO2,,;21,,211[6],:NADHyFlavinyFe-SFdyFe-SRyFeyO2Fe-SFdFe-SRRieske-[2Fe-2S]-Fe1Fe-S,Fe-S1(P.arvillaC-1,Acine-tobactercalcoaceticus)37,500A(50,000)3B(20,000)3FAD,2Fe-2S3[2Fe-2S],3FebenC(xylZ)benAB(xylXY)IB[7](P.putidaML2NCIB12190)A(42,000)211,860A(54,500)2B(23,500)2FAD[2Fe-2S]R2[2Fe-2S]R,FebedA(bnzC)bedB(bnzB)bedC1C2(bnzAB)IIB[8](P.putidaF1)46,00011,900A(52,500)2B(20,800)2FAD[2Fe-2S]R2[2Fe-2S]R,FetodAtodBtodC1C2IIB[9](P.putidaNCIB981)36,30015,300A(55,000)2B(20,000)2FAD,[2Fe-2S][2Fe-2S][2Fe-2S],2FendoAndoBCIII[10](P.spLB400)A(51,000)3B(22,000)212,000,43,000[2Fe-2S]FebphAbphEbphFbphGIIB[11](Ñ)Fe-S,(FMN)IA,(FAD)IB(ben-dox);(Ò)Fe-S(ben-dox)(to-ldox);(Ó)Fe-S,Ó(nap-dox)4321111(FerredoxinReductase),,NADH,42~67kDa,Fe-S,,,38~44kDa[2Fe-2S]N-FMN,;C-21112[2Fe-2S](Ferredoxin),,[2Fe-2S],[2Fe-2S]2,2,44,Fe(Ó)Fe(Ò)[2Fe-2S]RieskeJ.S.RieskeRieske[2Fe-2S]Gibson,21113(Iron-sulfurPro-tein,ISP),,O2Fe2+,,Rieske[2Fe-2S],150~200kDa,5020kDaABA2B2A3B3ARieske[2Fe-2S],BAB212:-2,3-(C23O)[12]432kDa,Fe2+,2-,xylE:,O2,,Fe3+,-1,2-(C12O)[13]AB,[AB-Fe3+]n,[AA-Fe3+]n3311P.putidamt-2[14],TOLpWWO,TOL,,xylCBA,xylDLEGFJKIHbphtol[15]bphtol,bphAORFto-ldoxP.putidaKF707bphD()bphXtolF()tolCl2,3-60%(BphDTolF),35%BphDTolFP.putidaNClB8164P.putidaG7(nah)[16],,16kbnahnahnahR,,nahna-hAaAbAcAd44,Barkholderiasp.RP007[17]PAH,312TOL[18],xylRxylSXylR,,PuXylSPm,Pm,XylRPs,XylS,PmXylS,XylSPmXylRPs&Pu313,[19],LeetodC1C2BA,P.putidamt-2,P.putidaB13[20]:3-4-,24-(2or4-CBA)3,5-(3,5-DiC-BA),P.putidaB13,4,,[21],[22],(to-ldox)0107401091,,C23O,0183,C23O[1]ShiraiKetal.,1987.Agric.Biol.Chem.,51:121-128.[2]SmithMRetal.,1989.Appl.Microbiol.Biotechnol.,32:68-75.[3]SariaslaniFSetal.,1974.Biochem.J.,140:31-45.[4]SmithMRetal.,1989.Appl.Microbiol.Biotechnol.,30:395-401.[5]CarnigliaCE,1984.Adv.Appl.Microbiol.,30:31-71[6]MasonJRetal.,1992.Annu.Rev.Microbiol.,46:277-305.[7]NeidleELetal.,1991.J.Bacterial.,173:5385-5395.[8]GearyPJetal.,1990.MethodsEnzymol,188:52-60.[9]HaiyanJetal.,1999.Appl.Enviroment.Microbiol.,65:315-318.[10]MichealJLetal.,1999.J.Bacterial.,181:6200-6204.[11]JohnDHetal.,1995.J.Bacterial.,177:5834-5839.[12]HugoNJetal.,1998.J.Bio.Chem.,273:9622-9629.[13]SendaTKetal.,1996.J.Mol.Biol.,255:735-52.[14]HarayamaSetal.,1986.J.Bacterial.,167:251-255.[15]FurukawaKetal.,1993.J.Bacterial.,175:5224-5232.[16]YenMYetal.,1988.Crit.Rev.Microbiol.,15:247-268.[17]AndrewDLetal.,1999.J.Bacterial.,181:531-540.[18]HoltelAetal.,1990.Mol.Microbiol.,4:1551-1556.[19]LeeJYetal.,1995.Appl.Enviroment.Microbiol.,61:2711-2717.[20]RojoFetal.,1987.Science,238:1395-1397.[21]ReganLetal.,1993.ComputersChem.Engng.,17:627-637.[22]RoseSetal.,1998.Biotechnol.Bioengin.,58:240-249.45AdvanceontheStudiesofBiodegradationofAromaticcompoundsSunYanQianShijun(InstituteofMicrobiology,AcademiaSinica,Beijing100080)AbstractBiodegradationofaromaticcompoundssuchasbenzenealkylbenzenesbiphenylandpolycyclicaromatichydrocarbonsisreviewed.Theconversionofthemtocis-diolsandthering-cleavagebytwokindsofdioxygenasesareallinvolvedinthepathway.ThemolecularbiologyresearchabouttheTOL-pathwayisalsodis-cussedhere.Metabolicengineeringisintroducedintothearomaticcompoundsdegradationinearly90.s.Inthepathway,rate-controllingenzymecanbedeterminedandthefluxthroughthepathwayincreasedbyincreasingthetiteroftherate-controllingenzyme.KeywordsAromaticcompound,Biodegradation,Dioxygenase,Molecularbiology,Metabolicengineering(58),,19979,FDA,,90,,214/0,,,,100%,300%,3311,,,100,1/50,312,,,,313,,314,,/0/0315,,,,[1],,561999[2]EllenLickinget.al,BusinessWeek/China,2000.5:6746