摘要
目的:分离纯化苦瓜溶菌酶,同时进行酶学性质及其生物学性质研究,为苦瓜生物活性蛋白的研究提供依据。方法:应用盐提法、离子交换层析法(阴离子色谱柱Source Q和阳离子色谱柱POROS 50HS)、反相毛细管色谱和SDS-PAGE等方法分离纯化和鉴定。结果:分离纯化得到的溶菌酶经鉴定达到电泳纯,分子质量为20.1kDa,酶比活力2000U/mg,纯化倍数164.6;抗菌试验表明,苦瓜溶菌酶对总状毛霉(M ucor racem osus)、稻纹枯病菌(osus Rhizoctonia solani)、大肠杆菌(Escherichia coli)具有较强的抑制作用,对大肠杆菌完全抑制的最低浓度为0.59μmol/L,而对金黄色葡萄球菌(S taphy lococcus aureus)的抑制活性不明显。结论:分离表征到一种具有抗菌活性的新型植物溶菌酶,使用离子交换层析柱高效可行。
Objective： To isolate and purify a lysozyme from Momordica charantia L and study the enzymatic and biological characterizations of the lysozyme. Methods： salt extraction, ion exchange chromatography on high-performance liquid chromatography （HPLC） on Source Q and Poros 50 HS, reversed phase-HPLC and SDS-polyacrylamide gel electrophoresis were used for isolation, purification and identification. Results： The lysozyme showed a molecular mass of 20.1 kDa in SDS-polyacrylamide gel electrophoresis. Compared with the crude extract, the resulting enzyme solution was purified by 164.6 folds, and its specific activity was 2 000.0 U/mg. The lysozyme exerted antifungal action toward M ucor racem osus and osusRhizoctonia solani, and antibacterial action against Escherichia coli and S taphy lococcus aureus. The minimal inhibitory concentration against Escherichia coli was determined as 0.59 μmol/L, while its antibacterial action against S tapy lococcus aureus was insignificant. Conclusion： The method presented in this paper was feasible to purify the lysozyme from Momordica charantia L, and the newly purified lysozyme was proved to be a novel plant lysozyme.
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2009年第5期55-60,共6页
Journal of Chinese Institute Of Food Science and Technology
基金
福建省科技厅重点项目资助(2008-2011)
关键词
苦瓜
纯化
溶菌酶
抗菌蛋白
Momordica charantia L
Purification
Lysozyme
Antibacterial and antifungal protein
