CNBr-Papain-Sepharose 4B亲和填料的

doi:10.3969/j.issn.1673-1689.2021.02.013

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CNBr-Papain-Sepharose 4B亲和填料的
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                        10.3969/j.issn.1673-1689.2021.02.013
                    
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Preparation and Properties of CNBr-Papain-Sepharose 4B Affinity Media

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为探求从鱼类废弃组织及漂洗水中回收Cystatin的高效方法，首先自制了溴化氢-木瓜蛋白酶-琼脂糖凝胶（CNBr-Papain-Speharose 4B）亲和填料，确定了适宜的配基Papain偶联密度，之后测定了该亲和填料的热稳定性、酸碱稳定性、贮藏性能及使用的重复性，最后以该制备的填料对鱼糜漂洗水中的CPIs活性进行了初步回收。以对重组鲢鱼Cystatin吸附率为评价指标，确定适宜Papain偶联密度，即CNBr-Speharose 4B和Papain溶液最佳料液比为1∶7（g∶mL）。CNBr-Papain-Speharose 4B的适宜使用温度为4~40 ℃，且在pH 3.0~12.0之间稳定。CNBr-Papain-Speharose 4B在4 ℃下，6个月的贮藏期内，表现较好的稳定性。将填料装柱后进行验证实验，证明该填料的使用重复性能稳定，且最终成功高效地回收了鲢鱼鱼糜漂洗水中的CPIs。

Abstract:

To explore an efficient method of recycling Cystatin from fish waste tissue and rinsing water, the self-made affinity media CNBr-Papain-Speharose 4B was firstly prepared. The optimal coupling density of Papain was further determined. Then the thermal stability, acid-base stability, storage performance and repeatability were respectively detected. At last, the active CPIs in silver carp surimi rinse water was preliminary recycled by the self-made affinity media. The optimal coupling density of Papain was evaluated by the adsorption rate of recombinant carp cystatin. The best liquid-solid ratio of CNBr-Speharose 4B toPapain was indicated as 1∶7(g∶mL). The optimal temperature of CNBr-Papain-Speharose 4B was 4~40 ℃ during usage, and it was stable between pH 3.0~12.0. The storage detection suggested that CNBr-Papain-Speharose 4B was stableat 4℃ within 6 months. The affinity media's repeatability was verified to be stable. Finally, CNBr-Papain-Speharose 4B affinity media successfully recycled CPIs from silver carpsurimi rinse water with high efficiency.

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刘明宇,李树红,杨娟,林昊,杨珂,谭小千. CNBr-Papain-Sepharose 4B亲和填料的[J].食品与生物技术学报,2021,40(2):94-102.

LIU Mingyu, LI Shuhong, YANG Juan, LIN Hao, YANG Ke, TAN Xiaoqian. Preparation and Properties of CNBr-Papain-Sepharose 4B Affinity Media[J]. Journal of Food Science and Biotechnology,2021,40(2):94-102.

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在线发布日期: 2021-05-28
出版日期: 2021-02-15

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