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1、短短芽砲杆菌JK-2胞外多糖分离与纯化条件优化及成分鉴定张秋芳,杨述省,史怀,彭奇S刘波*(1福建省农科院生物技术研究所,福建福州350003;2中国科学院福建物质结构研究所,福建福州350002)摘要:本研究对JK-2胞外多糖提取和分离条件进行优化,并采用柱层析、薄层层析及红外光谱等方法,确定其组成分。将浓缩上清液加入乙醇至18.75%,13000rpm离心16minJu,再加终浓度为60%乙醇,离心得粗多糖;加入氯仿:正丁醇体积比为2:1、提取4次、振摇10min,粗提液:捉取剂体积比1:2去除粗蛋白后,透析;用SephacrylS-200和S
2、uperdex-75柱进行分离,皆出现明显单峰;分子量为7.868X10*03,由单一组分D■果糖组成;含有竣酸、毗喃单体,但无乙酰酯。关键词:胞外多糖;短短芽砲杆菌JK・2;分离纯化;条件优化;成分鉴定IsolationandpurificationsofconditionoptimizationandingredientidentificationofextracellularpolysaccharidefromBrevibacillusbrevisJK-2ZHANGQiu-fang,YangShu-sheng,ShiHuai,PengQi2a
3、ndLIUBo"(BiotechnologyInstitute,FujianAcademyofAgriculturalSciences,Fuzhou,Fujian350003,China;2.FujianInstituteontheStructureofMatter,ChineseAcademyofSciences,Fuzhou350002,China)Abstract:IsolationandpurificationsofconditionoptimizationofextracellularpolysaccharidefromBrevibaci
4、llusbrevisJK-2wereconducted.Anditsingredientidentificationwasalsotestedbythecolumnchromatography,thethinlayerchromatographyandtheinfraredspectrum.Thesupemateoffermentwhichhadbeenfiltratedandconcentratedwasaddedtheethanolupto18.75%,13000rpm,16min.andthenaddedethanolreach60%,The
5、centrifugalresultsgainedthecrudepolysaccharide;Withthechloroformvolumeastwotimesasthebutanol,extracted4times,swayed10mineachtime,thecrudepolysaccharidewasremovedthecrudeprotein;andthendialyst;ItCarriesontheseparationwithSephacrylS-200andtheSuperdex-75gelatincolumn,allappearedo
6、bviouslysinglepeak,themolecularweightofitisabout7.868x104DaandcomposedofD・Fructose,carboxylicacids,pyranemonomer,butnotperoxyacetylgroup.Keywords:Exotracelluarpolysaccharide(EPS);BrevibacillusbrevisJK-2;Isolationandpurification;Conditionoptimization;Ingredientidentification基金项
7、目:福建省财政专项■福建省农业科学院科技创新团队建设基金(STIF-Y03),福建省H然科学基金资助项目(B0410024);福建省农科院预试课题(A2006YYS09)作者简介:张秋芳,女,硕士,副研究员,主要从事植物营养生理及生化物质分析等方而研究。E-mail:qfzhang@163.com□"通讯作者:刘波,男,博士,研究员,主要从事生物技术和生物防治研究,E-mail:fkliubo@163.com。刖s随着糖类研究的深入,大量实验结果揭示糖类是携带着最大信息量的一类具有生理活性的高分子物质。细菌胞外多糖(Exotracelluarpol
8、ysaccharide,EPS),是指细菌在生长、代谢过程屮分泌到细胞外的粘液或夹膜多糖。其成分和结构复杂,具有调节和增强