淀粉酶纳米花型时间
摘要： 目的：研制基于蛋白质杂化纳米技术和酶促反应的淀粉酶纳米花型时间-温度指示剂（time-temperature indicator，TTI），确定其活化能及适合指示的食品种类。方法：利用淀粉酶与铜离子之间相互作用，对淀粉酶进行固定化，形成淀粉酶@Cu纳米花，改变淀粉酶用量，筛选出花形最好的纳米花，并测试纳米花中淀粉酶的生物活性及保存稳定性；以15 mL 40 g/L可溶性淀粉溶液为底物，4.5 mL 1 g/L碘液为指示剂，分别加入10、20、30、40、50、60 mg的淀粉酶@Cu纳米花，制备出6 种配方的TTI；在5、15、25、35 ℃的恒温条件下，研究6 种TTI的性能，观察变色过程，测定动力学参数，计算反应活化能值。结果：淀粉酶质量浓度为0.5 mg/mL时，淀粉酶@Cu纳米花的花形最完整，淀粉酶活性提高到3.42 倍，保存稳定性显著提高；6 种TTI呈现出从深蓝色到无色的变化过程，活化能分别为14.84、21.00、28.85、33.03、32.55 kJ/mol和32.83 kJ/mol。结论：淀粉酶纳米花型TTI操作简单，指示效果明显，根据TTI与食品匹配原则，该系列TTI适用于监测由扩散控制、酶反应、脂肪氧化等原因引起腐败变质的食品质量变化。
关键词: 时间-温度指示剂；淀粉酶；纳米花；吸光度；活化能
Abstract: Objective: To develop a time-temperature indicator (TTI) with amylase nanoflowers based on protein hybrid nanotechnology and enzyme reaction and to determine the activation energy for TTI and its suitability for different food types. Methods: Based on the interaction between amylase and copper ions, amylase was immobilized to form amylase@Cu nanoflowers. The amylase concentration for nanoflowers with the best shape was selected, and the biological activity and stability of amylase in the nanoflowers were tested. Six formulations of TTI were prepared by adding 10, 20, 30, 40, 50 mg and 60 mg of amylase@Cu nanoflowers and using 15 mL of 40 g/L soluble starch solution as the substrate and 4.5 mL of 1 g/L iodine solution as the indicator. At constant temperatures of 5, 15, 25 and 35 ℃, the discoloration process was observed, the kinetic parameters were measured, and the activation energy for the reaction was calculated. Results: When the amylase concentration was 0.5 mg/mL, the flower-like shape of amylase@Cu nanoflower was the most complete, the amylase activity increased by 3.42 times, and the storage stability was significantly improved. The six TTIs showed a change process from dark blue to colorless, and the activation energy values were 14.84, 21.00, 28.85, 33.03, 32.55 and 32.83 kJ/mol, respectively. Conclusion: The amylase nanoflower-based TTI was simple to operate and had good indication performance. According to the matching principle between TTIs and foods, this series of TTIs were suitable for monitoring food deterioration and spoilage caused by diffusion control, enzymatic reaction, fat oxidation or other reasons.
Key words: time-temperature indicator; amylase; nanoflowers; absorbance; activation energy
中图分类号:
TS207.7
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