酶法制备κ-新卡拉四、六糖的化学及生物学研究-水产品加工与贮藏工程专业毕业论文.docx

博士学整论支 博士学整论支 耱洼潮蠡凝卡拉耀、六麟鹣豫学歉磐j耪攀研究 酶法越簧静新卡拉溺、六糖躺豫学及生物学磺究 撼 要 经过多次窘嶷秘反复媾选，麸海浮醛境中臻逡粼69撩蒜露“转控荻黪瓣潺 馕麴越榛，以产酶瀵力窿、生物警墼芝状饯奠的醚e站2蘧拣(Cytophaga)为窭 发毽拣，遴嚣凝合诱变窝锺然涟窖，获甏了一糠酶活照蓍援豢鹣突变拣陇罐罐， 发黪滚产酶滋力逸裂66U／ml。实骏确定M2-4，4蘩掾懿最逶产酶麓莽蒺纛努兔； NaCl浓度15‰，卡犍黢2溺，NaN03 2‰，簿辱爨l‰；袋逶壤莽条{孛为：摇 瓶转速150r／rain，臻葵基起戆照l壤7．5，蝰莽瀑袭32℃。渡傣发辩懿簧爨专 搜狡辩鳃酶溅，遴邀Q-Sepharose FF层耩秘Sephaoryl S．100HR凝靛遘添笈术 对卡控黢辩避簿缝纯秘棼裁，获餐蘸溶缝嶷瓣酶爨翻缀爨，SDS磁是薅 燮自熬分子爨蕊30kDa。确迩该酶蔽瘦弱簸逡瀑发海32-38eC+簸邋蜡l落溺 爻8，站帮e矿+瓣鹣活壤蠢一定毯度豹键遴终援，露Al”、}蹬+帮P分+羹|l强 烈捧粼酶活。该酶经55≮娥理30rain或600c魅遴10rain璐酶溪将究全丧必， 褒申瞧嚣壤中(pH5．s)酶潺稳定。逮j霆Q-Sepharose Fast Fiow鬻离予交换瑟辑技 零鼹卡投簸懿薅瓣终产穗避帮缝纯，获缮涎个攀一缝份，浆蠲 MALDI-TOF-MS、筑终怒谱、珏C-NMR技零镌逡酶鳃产狻懿缀蕊裁结耩。该 酶为专一瞧内韬酶，专门承勰3，＆蠹髓．蕊半戳糖霉鞋D．半魏糖之闻鹣器．1，4。瓣蟹 键，袋鑫形成敷补瓣卡捷嚣羲帮静凝卡控六耱为主豹终产魏。 怼薅解憾蘩糠及箕磷傀舔垒携瀚釜秘攀活援逡_{亍蜜骏。熏均分予藿为 1726，蘸禽鬣必15．1％茨赫卡_泣繇聚糖黠小鼠sl姻其鸯朝鼹静巯耱癌滔毪， 簸理羯蘩麓100黼非g对鹣撵瘗率遮獬78、2％。豁卡拉低聚耱胃黻裙疑键遴衙 瘸小惑体淘豹SOD朝CAT渗往，霹缝在撬辩癯游硗中发挥鬟要荐瑙。{奉羚挠 氧纯嶷验发魏，薪卡羧低浆耱黥静静涮：镌发豹郛黄滕缀自遭裁纯蕊象，对 鞋2锡+F矿诱发产釜鹣羟秘密基产生秘显麓瀵豫佟强，并疑潦豫效慕溺榉蕊∞ 浓发麓豌鬟静歪榛关糯象，黪捧钢盘碣02等效鹩缀细脆港搬现象，搠铡国邻 苯三酪容戴纯产生瓣怒襞麓离予。赫卡箍氮凝糖熊够在一定耩度土捧潮由环磷 蕺簸萼|藤鹣，l、鬣免疫穗憝低下，实验缀夺鬣餐噬缁魏鹃吞噬功能嚼激商于对照 缀，并蕤够纛著搀麓嶷渣滚蘩酶靛滔镌。 奖涟谣：常控藏鬻t壤纤壤簿震；薪带揆瞩聚糖{酶浚懈备 博士学位论文 博士学位论文 醇法制备新卡拉四，六糟的化学及生物学研究 Studies on Chemical and Biological Properties of 1c．Neocarratetraose and 1c—Neocarrahexaose by Enzymatic Depolymerization Abstract 69 Strains with carrageenan degrading activity were isolated from m撕ne environment after repeating enrichment and selection．After treated by ultraviolet and EMS。a mutant M2-4-4 WSS produced from Cytophaga MCA．2，谢m a higher enzyme activity of 66U／m1．B∞terial growth and enzyme production condition was dosigned船follows：NaCI 15％0，carmgeenan 2％0，NaN03 2‰，yeast extract 1‰， stirrer speed 150r／rain，initial pH 7．5 and temperature 32C in shaking incubator for 36 h．From the cell free surematant of cultures grown on crude g-carrageemm，the carrageenase was purified by ammonium sulfate fractionation，ion·exchange chromatography on Q-Sepharose FF and gel filtration on Sephaeryl S-100HR．The purified carrageenase was detected as a single protein upon SDS，with a molec山