转谷氨酰胺酶交联酶晶体的制备及其结构表征  

Preparation of Cross-linked Transglutaminase (MTG) Crystals and its Structure Characterization

作　　者：韩春然[1] 张娜[1] 王强[1] 张毅方 刘颖[1] 马永强[1] 

机构地区：[1]哈尔滨商业大学食品工程学院省高校食品科学与工程重点实验室,哈尔滨150076 [2]哈高科大豆食品有限责任公司,哈尔滨150078

出　　处：《中国食品学报》2011年第4期42-47,共6页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家高技术研究发展计划(863计划;2008AA10Z303);黑龙江省高校科技创新团队建设计划(No.2010td04)资助

摘　　要：采用双功能试剂戊二醛对转谷氨酰胺酶晶体进行化学交联,得到交联条件。对交联酶晶体用电镜扫描以及红外光谱和XPS进行结构表征。交联条件:戊二醛质量分数1%,交联pH6,交联温度4℃,交联时间40min。红外光谱显示,游离酶在1651cm-1出现强峰,而交联酶晶体在1634、1281和1241cm-1出现峰值,在α-螺旋区域没有检测到峰值。通过X-射线光电子能谱分析证实,交联酶晶体与游离酶相比,在285.1eV处的C1S的信号峰增强,在400.375eV处的N1S峰几乎消失,说明酶蛋白表面的游离氨基已经与戊二醛结合,表面蛋白结构发生了变化。Transglutaminase（MTG） crystals was cross-linked chemically by bifunctional reagent——glutaric dialdehyde,the cross-linking conditions were obtained,and the cross-linked enzyme was scanned by scanning electronic microscope and its structure was characterized by infrared spectrum. Cross-linking conditions were as follows,glutaric dialdehyde 1%,pH 6,4 ℃ for 40 min. The scanning picture showed that when the free MTG cross-linked into crystals,the space molecular structure changed,the size of the cross-linked crystals were uniform and anomalous polyhedron. The infrared spectrum showed that,strong peak appeared on 1 651 cm-1 in free MTG,while the strong peak appeared on 1 651 cm-1 in cross-linked crystals,peaks appeared on 1 281 cm-1 1241 cm-1 in cross-linked MTG,respectively,while there was no peak in alpha helix region.

关 键 词：转谷氨酰胺酶 交联 酶晶体 表征 

分 类 号：TS201.21[轻工技术与工程—食品科学]