微生物酶法拆分2芳基丙酸类药物研究.pdfVIP

南开大学硕士研究生毕业 (学位)论文 ABSTRACT ， Seventeen bacterial strains were cultured from the soil in the vicinityofN即roxenproducingfactory,fromwhich14strainspossessed lipase activity. A Bacillus strain was screened for asymmetric resolutionof (劝-Naproxen.Thisstrainwasstudiedforitsculture conditionsofgrowthandlipaseproduction,itsconversionconditionsof 2-Arylpropionicacids,thepositionsandthepropertiesoflipasefrom itsbacterialcell. Experimentalresultsshowedthattheopticalpurity (ee%)of (劝- Naproxenwasover89%andconversionratewasover30% inbacterial cellsPBSreaction system.After theBacillusbacterialcellswere processedbylysozymeandquartzsand,dissolvedlipasewasclarifiedby centrifugationand loadedonaphenyl-SepharoseCL-4B column.After purification.bya singlehydrophobic chromatography, thepurity of lipaseincreased330timesandtheactivityoflipasewasapproximately 43timeshigherthanthecrudeone.Thehydrolyticactivityoflipase usingNaproxenethylesterandpNPAassubstrateremainedessentially constant with bacterial cells PBS reaction system 山ring the purificationprocedure. D主fferent lipases from the eluting fractions by hydrophobic chromatographyWere purifiedfurtherbypreparative electrophoresis. Threelipasesof97KD 55KDand40KDwereobtainedwithhighpurityand the activitv of conversion ofNaproxen ethylester. isoelectric focusingelectrophoresispreliminarilyrevealedthattheplofthree purifiedlipasesof97KD,55KDand40KDwere5.7,7.3 and4.9一 5.7, respectively. Keywords:lipase;(劝-Naproxen;asymmetricresolution;hydrophobic chromatography;preparativeelectrophoresis 南开大学硕士研究生毕业 (学位)论文 前 言 对映体的存在是自然界中的一种普遍现象，在有机化学、药物化学和生物化学 等领域尤为突出u〔。对手性药物而言，并非两种异构体都具有相同的活性和药理特 征。例如， “心得安”的(S)一(-)异构体的疗效是(R)一(+)异构体的100倍;反应停 (Thalidomide)是一种外消旋的手性药物，其(R)一异构体具有镇静作用，而((S)- 异构体却具致畸作用，二十世纪50年代妊娠妇女服用此药后，多例出现了畸变胎