摘要
目的:分离纯化鱿鱼眼透明质酸,为利用鱿鱼加工废弃物提供理论依据。方法:以北太鱿鱼眼为原料,通过正交试验考察酶解时间、温度、酶用量对透明质酸提取率的影响,优化酶法提取透明质酸工艺条件;以离子交换层析分离纯化,用紫外光谱和凝胶过滤层析鉴定透明质酸的纯度及其分子质量,并用红外光谱分析其结构。结果:1)枯草杆菌蛋白酶提取透明质酸的最适条件为酶解时间1h,温度60℃,酶用量4%,鱿鱼眼透明质酸的提取率达14.10%;2)利用H2O和NaCl溶液分步洗脱,DEAE-Sephadex A-25离子交换层析纯化得到2个透明质酸组分HA-1和HA-2,得率分别为5.22%和84.37%;3)HA-1和HA-2组分的相对分子质量分别为6.77×105、1.73×106,均显示为单一洗脱峰,无蛋白、核酸杂质;红外图谱显示其具有透明质酸标准品的吸收峰。结论:鱿鱼眼透明质酸酶法提取率14.10%,纯化得到的透明质酸分子质量分别为6.77×105和1.73×106。
Extraction technology of hyaluronic acid（HA） from northern pacific squid eyes was developed,the effect of zymolysis time,zymolysis temperature and the dose of enzyme on the yield of hyaluronic acid was examined by orthogonal test.The HA was purified by ion exchange chromatography,then the purity and molecular weight of HA were iden-tified by UV spectrum and gel-filtration chromatography.In addition,the composition analysis was studied by IR spectrum.The optimum technology was as follows：zymolysis time 1h,zymolysis temperature 60 ℃ and the dose of enzyme 4%.On the optimal condition,the yield of HA was 14.10%.As a result,loaded on a DEAE-Sephadex A-25 column and eluted with H2O and 0.95 mol/L NaCl in turn,two kinds of hyaluronic acid marked HA-1 and HA-2 were separated and their yields were 5.22% and 84.37%,respectively.Both of the two components had no protein and nucleic acid left and each of their purity was shown to be on a single elution peak.The average molecular weight of HA-1 and HA-2 were 6.77×105 and 1.73×106,respectively.The infrared spectrum of HA-2 was accordance with that of standard HA.
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2010年第4期88-93,共6页
Journal of Chinese Institute Of Food Science and Technology
基金
农业部948项目(2006G-41)
浙江省新苗人才计划项目(2008R40G2070044)
关键词
鱿鱼眼睛
透明质酸
酶法提取
纯化
squid eyes
hyaluronic acid
enzymatic extraction
purification
