智能凝胶载体固定化纤维素酶提取植物活性成分  

Extraction of Active Components from Plants Using Cellulase Immobilized on an Intelligent Gel Support

作　　者：周建芹 陈韶华[2] 

机构地区：[1]苏州大学药学院,江苏苏州215123 [2]苏州大学医学部实验中心,江苏苏州215123

出　　处：《华南理工大学学报（自然科学版）》2012年第1期125-130,137,共7页Journal of South China University of Technology(Natural Science Edition)

基　　金：江苏省科技支撑(社会发展)计划项目(BE2010649);江苏省自然科学基金资助项目(BK2010225)

摘　　要：为提高植物活性成分的提取率,同时实现酶的重复利用,以智能凝胶材料N-琥珀酰壳聚糖(NSC)为载体固定化纤维素酶.文中首先采用红外光谱仪和X射线衍射仪对NSC的结构进行了表征,并研究了其pH敏感性.随后探讨了NSC固定化纤维素酶(NSCC)的制备条件;最后以银杏叶为模式材料,用NSCC水解提取黄酮类化合物,并确定了NSCC水解的最适温度和pH值.结果表明:琥珀酸酐与壳聚糖中的氨基反应生成了NSC,且琥珀酸酐的引入使壳聚糖中的结晶区减少;NSC具有对溶液pH值敏感的可逆溶解性,NSCC也表现出类似的pH敏感性;NSCC的最优制备条件为pH 5.0、1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐用量10 mg、纤维素酶用量15 mg;NSCC水解能显著提高银杏叶粉中黄酮类化合物的提取率,提取率仅略低于游离酶,且NSCC可以重复使用;NSCC水解银杏叶粉的最适温度是45～55℃,最适pH值是5.0.In order to enhance the extraction of active components from plant materials and realize the reuse of enzyme,an intelligent gel,namely N-succinyl-chitosan（NSC）,was used as a support to immobilize cellulase.In the investigation,first,the structure of NSC was characterized via the FT-IR and the XRD,and the pH sensitivity of NSC was also discussed.Then,the conditions of NSC-immobilized cellulase（NSCC） were optimized.Finally,the extraction of flavonoids from Ginkgo biloba leaf powders was carried out in the presence of NSCC,and the optimal temperature and pH value of NSCC for the extraction were determined.The results indicate that the production of NSC is due to the reaction of succinyl groups and the —NH2 groups in chitosan,that the crystalline region of chitosan becomes small due to the introduction of succinyl groups,that both NSC and NSCC possess similar pH-sensitive reversible solubility,that NSCC can be obtained under the optimal immobilization conditions of pH 5.0,10 mg of 1-（3-dimethylaminopropyl）-3-ethylcarbodiimide hydrochloride（EDC） dosage and 15 mg of cellulase do-sage,that NSCC with good reusability remarkably improves the extraction yield of flavonoids from Ginkgo biloba leaf powders,which is only a little less than that by using free cellulase,and that the optimal temperature and pH value of NSCC for the extraction are respectively 45~55 ℃ and 5.0.

关 键 词：N-琥珀酰壳聚糖 纤维素酶 固定化 活性成分 提取 

分 类 号：Q814[生物学—生物工程]