重组梅花鹿来源Cu/Zn-SOD的分离纯化及酶学性质  

Purification and Characterization of Recombinant Cu/Zn-Cervus nippon Superoxide Dismutase

作　　者：许鑫琦 吕橄 李仁宽[1] 叶秀云[1] 林娟[1] Xu Xinqi;Lu Gan;Li Renkuan;Ye Xiuyun;Lin Juan(Fuzhou University, Fujian Provincial Key Laboratory of Marine Enzyme Engineering, Fuzhou 350116)

机构地区：[1]福州大学福建省海洋酶工程重点实验室,福州350116

出　　处：《中国食品学报》2018年第5期137-143,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：福建省高校产学合作重大项目(2013N5005)

摘　　要：超氧化物歧化酶(SOD)能特异性清除氧自由基,对保护细胞免受氧化损伤具有重要作用。本文构建了表达重组梅花鹿来源Cu/Zn-SOD的毕赤酵母工程菌GS115-p PIC9K-SOD,并对重组SOD进行分离纯化和酶学性质研究。通过10 ku中空纤维柱超滤、DEAE弱阴离子交换层析、高效凝胶色谱分离纯化,重组Cu/Zn-SOD的比活力为1 7647.1 U/mg,回收率为36.84%。对重组Cu/Zn-SOD的酶学性质研究表明,其最适反应pH 8.0,最适反应温度30℃;在pH 5~9之间较为稳定,保温1h后仍能保留80%以上的酶活力;60℃保温1 h酶活保持在90%以上,70℃保温20 min残余酶活力仍有70%以上。重组Cu/Zn-SOD在中性条件下对蛋白酶耐受性较好,然而易受胃酸的酸解导致酶活力丧失。Sika deer（Ceruse nippon） superoxide dismutase（SOD） is helpful for human health. In this paper, of Cervus nippon superoxide dismutase（SOD） was recombinantly expressed in Pichia pastoris strain GS115 and then determined enzymatic properties. The recombinant Cu/Zn SOD was purified through ultrafiltration and weak anion exchange chromatography（DEAE）, gel filtration to obtain specific activity of 17 647.1 U/mg. Research on the enzymatic properties showed that the optimum reaction pH of the SOD was about 8.0 and the optimal reaction temperature was 30 ℃. The SOD was relatively stable at pH 5-9 and below 70 ℃. Simulation study of human internal digestion found that this recombinant SOD was well tolerant for protease under neutral conditions but susceptible to gastric acid solution with severe loss of enzyme activity due to acid effect. This indicated that stomach acid is the main problem for oral intake of the SOD. This study is proposed to be basis for utilization of recombinant Cervus nippon SOD.

关 键 词：Cu/Zn-SOD 梅花鹿 毕赤酵母 纯化 酶学性质 

分 类 号：TS201.25[轻工技术与工程—食品科学]