Effect of incubation on stabilization of lipase immobilized by epoxy resin
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摘要:目的
研究甘氨酸孵育对LXEP-120环氧树脂固定化脂肪酶稳定性的影响。
方法使用甘氨酸孵育环氧树脂固定化脂肪酶，消除固定化酶载体上过剩的环氧基团，对孵育的条件进行探索和优化，并且比较孵育前后固定化脂肪酶的酶学性质。
结果孵育环氧树脂固定化脂肪酶的最佳条件为：选用浓度为2.5 mol/L、pH7.0的甘氨酸溶液，在25 ℃条件下孵育24 h。孵育后的固定化脂肪酶的相对酶活力在80 ℃条件下处理6 h后仍保存60%左右，而相同条件下处理后的未经孵育的固定化脂肪酶的相对酶活力只剩下45%左右；与未孵育的固定化脂肪酶相比较，孵育后的固定化脂肪酶最适反应pH(8.0)、最适反应温度(45 ℃)与孵育前相同，pH耐受性、操作稳定性和储藏稳定性变化趋势在孵育前后基本保持一致。
结论孵育消除固定化酶载体上剩余的环氧基团是必不可少的一个技术环节，甘氨酸孵育工艺可以较大程度地提高固定化脂肪酶的热稳定性，对反应pH、pH稳定性、操作稳定性和储藏稳定性等性质影响较小。
Abstract:ObjectiveTo study the effect of glycine incubation on the stability of lipase immobilized by LXEP-120 epoxy resin.
MethodGlycine solution was used to incubate lipase immobilized by epoxy resin for removing the residual epoxy groups. The incubation conditions were explored and optimized, and the enzymatic properties of the immobilized lipase before and after incubation were compared.
ResultThe optimal incubation conditions were 2.5 mol/L and pH 7.0 glycine solution incubating for 24 h at 25 ℃. Following incubation, the immobilized lipase still retained about 60% of the original activity after treatment at 80 ℃ for 6 h, while the unincubated immobilized lipase retained only about 45% of the original activity. The optimal reaction pH (8.0) and optimal reaction temperature (45 ℃) of the immobilized lipase after incubation were the same as those of the unincubated immobilized lipase, and the pH tolerance, operation stability, and storage stability were the same as those before incubation.
ConclusionRemoving residual epoxy groups on immobilized enzyme through incubation is one necessary technical step. Glycine incubation can greatly improve the thermal stability of the immobilized lipase with little influence on reaction pH, pH stability, operation stability and storage stability.
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Keywords:
- epoxy resin /
- lipase /
- immobilization /
- glycine /
- stability
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图 1 氨基酸溶液孵育对固定化脂肪酶活力的影响
Figure 1. Effect of incubation with amino acid solution on immobilized lipase activity
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图 2 不同浓度甘氨酸溶液孵育对固定化脂肪酶活力的影响
Figure 2. Effect of incubation with different concentrations of glycine solution on immobilized lipase activity
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图 3 不同pH甘氨酸溶液孵育对固定化脂肪酶活力的影响
Figure 3. Effect of incubation of glycine solution with different pH on immobilized lipase activity
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图 4 孵育温度对固定化脂肪酶活力的影响
Figure 4. Effect of incubation temperature on immobilized lipase activity
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图 5 孵育时间对固定化脂肪酶活力的影响
Figure 5. Effect of incubation time on immobilized lipase activity
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