| 摘要: |
| 以海洋脂肪酶ADM47601为研究对象, 以树脂为载体, 进行了固定化酶的研究。筛选了21种树脂, 确定D316型阴离子交换树脂为载体。通过正交试验得到了固定化的最佳条件, 每克载体加脂肪酶10 000 U, pH7.5, 25℃, 150 r/min, 固定化时间3 h, 固定化酶活力回收率可达65.53%±1.06%。固定化脂肪酶最适作用pH为7.5, 在pH6.0～9.0的范围内能够保持85%的活力。最适作用温度40℃, 在50℃下贮存1 h后依然保持66%的活力。25℃保存半衰期为55 d, 操作8次仍然能维持60%以上的活性, 且在多种有机试剂中能够稳定的保持活性。固定化脂肪酶Km和Vmax值分别为4.86×10-5 mol/L 和1.31×10-5 mol/(L·min), 对底物亲和性较好。 |
| 关键词: 脂肪酶 树脂 固定化 酶学性质 |
| DOI：10.11759/hykx20160717001 |
| 分类号: |
| 基金项目:国家自然科学基金-联合基金(U1406402-5); 国际科技合作与交流专项(2014DFG30890); 国家实验室-鳌山科技计划(2015ASKJ02-06) |
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| Research on immobilization of lipase ADM47601 from marine microorganisms by resin |
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SUN Guo-long,MA Zi-bin,HAO Jian-hua,SHENG Jun,SUN Mi
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| Abstract: |
| The effect of different resins on immobilizing lipase ADM47601 was investigated in this study. A total of 21 types of resins were screened, and the anion-exchange resin D316 demonstrated the best immobilization. After the orthogonal test, the recovery ratio of enzyme activity reached up to 65.53% ± 1.06% under optimal conditions: 10000U lipase per 1g resin, pH7.5, 25℃, 150r/min and 3 h for immobilization. The optimal pH of immobilized lipase was 7.5, and it exhibited 85% of the maximum activity in the range of pH6.0–9.0. The optimal temperature was 40℃, and the activity was maintained at 66% after 1 h of storage at 50℃. At 25℃, the half-life was 55 days and the residual activity was still higher than 60% after utilizing the lipase 8 times. The immobilized lipase can maintain activity in various organic reagents, and its Km and Vmax values were 4.86 × 10-5mol/L and 1.31 × 10-5mol/(L·min), respectively, indicating high affinity to the substrate. |
| Key words: Lipase Resin Immobilization Enzymatic property |
