| NAD-Sepharose 6MB亲和胶的制备及在2,5-二酮基-D-葡萄糖酸还原酶纯化中的应用 |
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| 引用本文： | 吉爱国,高培基.NAD-Sepharose 6MB亲和胶的制备及在2,5-二酮基-D-葡萄糖酸还原酶纯化中的应用[J].山东大学学报(医学版),2000,38(3):258-259. |
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| 作者姓名： | 吉爱国 高培基 |
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| 作者单位： | 1. 山东医科大学生化制药教研室
2. 山东大学微生物技术国家重点实验室 |
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| 摘 要： | 目的制备NAD亲和胶，用于分离纯化2，5-二酮基-D-葡萄糖酸还原酶。方法将NAD通过丁二酸二酰肼与溴化氰活化的Sepharose6MB胶联结，制得NAD亲和胶。从棒杆菌细胞中提取的2，5-二酮基-D-葡萄糖酸还原酶粗酶液用NAD亲和胶进行亲和层析纯化。结果NAD的结合率为71．67％；亲和层析回收率为77．27％；酶的比活提高了1．1倍。结论以NAD为配基的亲和胶用于纯化以NADPH为辅酶的2，5-DKG还原酶行之有效。
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| 关 键 词： | 色谱法，亲和 烟酰胺腺嘌呤二核苷酸 2，5-二酮基-D-葡萄糖酸还原酶 |
| 文章编号： | 1000-0496(2000)03-0258-02 |
| 修稿时间： | 2000年8月20日 |
PREPARATION OF NAD-SEPHARSE 6MB AND ITS APPLICATION IN PURIFICATION OF 2,5-DIKETO-D-GLUCONIC ACID REDUCTASE |
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| JI Ai-guo,GAO Pei-ji.PREPARATION OF NAD-SEPHARSE 6MB AND ITS APPLICATION IN PURIFICATION OF 2,5-DIKETO-D-GLUCONIC ACID REDUCTASE[J].Journal of Shandong University:Health Sciences,2000,38(3):258-259. |
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| Authors: | JI Ai-guo GAO Pei-ji |
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| Abstract: | Objective:To prepare NAD Sepharose 6MB affinity gel and investigate its utility in separation and purification of 2,5 diketo D gluconic acid (2,5 DKG)reductase. Methods:NAD was combined with Sepharose 6MB by succinate acyl hydrazides to get NAD Sepharose 6MB affinity gel.Crude 2,5 DKG reductase was separated from Corynebacterium sp.ATCC 31090. The prepared NAD affinity gel was used for purification of 2,5 DKG reductase from the crude enzyme. Results:The combination rate of NAD to Sepharose 6MB is 71.67%. The recovery of affinity chromatography is 77.27%.The specific activty of the enzyme after purification is 1.1 times higher than before. Conclusion:The NAD Sepharose 6MB affinity gel is effactive for purification of 2,5 DKG reductase. |
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| Keywords: | Chromatography affinity Nicotinamide adenine dinucleotide 2 5 diketo D gluconic acid reductase |
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