墨鱼缠卵腺糖蛋白提取工艺优化及纯化

Optimization of Extraction Process and Purification of Nidamental Gland Glycoprotein from Cuttlefish

本文以墨鱼缠卵腺为实验原料,通过单因素实验及响应面试验优化墨鱼缠卵腺糖蛋白的提取工艺,随后对最优工艺下得到的糖蛋白进行分离纯化,电泳分析其纯度,并进一步研究其蛋白及总糖含量。结果表明,墨鱼缠卵腺糖蛋白提取最佳工艺为提取温度24℃,提取时间3.6 h,NaOH浓度0.4 mol/L,料液比1∶20 g/mL,超声功率200 W。在此条件下进行验证试验,得出墨鱼缠卵腺糖蛋白实际得率为16.13%±0.16%;经DEAE-52纤维素离子交换柱层析、Sephacryl S-300HR柱层析分离后得到纯品糖蛋白,电泳分析表明所纯化的糖蛋白纯度达到电泳纯,其中蛋白含量为65.53%±0.8%,糖含量为31.74%±1.1%,得率为墨鱼缠卵腺去外层薄膜原料的0.91%。

The nidamental gland of cuttlefish was used as experimental material,and the single factor experiments and response surface test were used to optimize the extraction process of nidamental gland glycoprotein from cuttlefish.Then the glycoprotein obtained under that was separated and purified. And its purity was analyzed by electrophoresis,its protein and toted sugar content were follow researched. The results showed that the best extraction process of nidamental gland glycoprotein was:extraction temperature 24 ℃,extraction time 3.6 h,the Na OH concentration 0.4 mol/L,the ratio of material to liquid 1∶ 20 g/mL and the ultrasonic power 200 W.Under the condition of the verification tests,the actual yield of nidamental gland glycoprotein was 16.13%± 0.16%. Purified by DEAE-52 celloulus ion exchange column chromatography and Sephacryl S-300 HR column chromatography was to obtain pure glycoprotein.The electrophoresis analysis showed that the purity of the purified glycoprotein reached electrophoresis purity,and the protein content was 65.53%± 0.8%,and the sugar contents was 31.74%± 1.1%. The yield was 0.91% of the nidamental gland to the outer film material.