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1、第16卷第3期甘肃科学学报Vol.16No.32004年9月JournalofGansuSciencesSep.2004肺炎链球菌培养及其荚膜多糖的制备王剑虹,乔瑞洁,王欣茹,沈荣,赵萍(兰州生物制品研究所,甘肃兰州730046)摘要:肺炎链球菌荚膜多糖是肺炎疫苗的有效组分,采用自制培养基以15L立瓶和100L发酵罐对流行肺炎链球菌进行培养,探讨其生长和荚膜多糖产生情况1结果表明肺炎链球菌以上述两种方式大量培养均生长良好1采用7～9h的培养物上清液经超滤浓缩,以乙醇沉淀法制备荚膜多糖,所获多糖基本符合《欧洲药典》规定要求1立瓶培养的6B、
2、18C、19F及23F荚膜多糖产量分别达112mgöL、123mgöL、115mgöL及103mgöL,这些数据均高于欧洲专利Yavordios所述72mgöL的产量1用立瓶培养时多糖收量较低的1、5和14型(收量分别为40mgöL、43mgöL及25mgöL)肺炎链球菌进行发酵罐培养的多糖收量与立瓶培养的相似1关键词:肺炎链球菌;培养;荚膜多糖;纯化中图分类号:R962,R372文献标识码:A文章编号:100420366(2004)0320040205TheCultivationofStreptococcusPneumoniaeandP
3、reparationofCapsularPolysaccharideWANGJian2hong,QIAORui2jie,WANGXin2ru,SHENRong,ZHAOPing(LanzhouInstituteofBiologicalProducts,Lanzhou730046,China)Abstract:Thecapsularpolysaccharidewascurrentlyusedasaneffectiveantigeninpneumococcalvaccines.Streptococcuspneumoniaeserotypes1
4、,5,6B,14,18C,19Fand23Fwerecultivatedin15Lbottlesusingself2madeliquidmediumbyshaking;serotypes1,5and14Streptococuspneumoniaewerealsocultivatedina100Lbioreactorunderstirringconditionstoexplorethegrowthoftestedstrainsandtheproductionofcapsularpolysaccharide.Theresultsshowedt
5、hatthegoodbacterialgrowthwasobservedintheabovementionedcultivatedmodes.Thelogarithmicgrowthcanbeexhibitedaftercultivatedfor5～6hours.Thesupernatantsofculturewereusedtoextractpneumococcalpolysaccharidebyethanolprecipitation.Afterisolatingandpurifyingofbacterialpolysaccharid
6、e(PS),thequalitylevelsofthemostbacterialPSswerecoincidentwiththerequirementsofEuropeanPharmacopoeia.ThePSyieldsoftypes6B,18C,19Fand23Fwere112mgöL,123mgöL,115mgöLand103mgöLseparatively,whichhad1.52foldincreaseinyieldascomparedwiththosedescribedinthepublishedpatent[Yavordio
7、sandCousin(1983)EuropeanPatent0071515A1].ThePSsyieldsoftypes1,5and14strainsincultivationof15Lbottleswere40mgöL,43mgöLand25mgöLseparatively.ThePSsyieldsinbioreactorcultivationweresimilartothoseinbottlecultivation.Keywords:Streptococcuspneumoniae;cultivation;capsularpolysac
8、charide;purification肺炎链球菌(Streptococcuspneumoniae)感染是球菌性感染就占100万例1肺炎链球菌性肺炎的成导致人类发病和死亡的主要原因之一,最常见
