過氧化氫酶(catalase, CAT)在催化反應結束後有分離純化困難與重複使用率低的問題，為改善此缺點，本研究利用線性聚賴胺酸: l-PLL32與星狀聚賴胺酸:3s-PLL28、6s-PLL29來包覆過氧化氫酶，並以乳化反應與京尼平交聯反應製備成微米膠，再透過礦化作用在微米膠上沉析二氧化矽，形成包覆過氧化氫酶之二氧化矽/聚賴胺酸複合微米水膠。製備好之微米膠與包覆過氧化氫酶之二氧化矽/聚賴胺酸複合微米水膠會進行一系列結構鑑定。再進行酶之活性分析: 在包覆性上過氧化氫酶皆不會漏出；動力學分析中最大反應速率Vmax和米氏常數KM為包覆過氧化氫酶之六臂聚賴胺酸複合微米膠 (6s-PLL29/CAT)/ CTMOS=5% > 包覆過氧化氫酶之三臂聚賴胺酸複合微米膠(3s-PLL28/CAT)/ CTMOS=5% > 包覆過氧化氫酶之線性聚賴胺酸複合微米膠(l-PLL32/CAT)/ CTMOS=5%；循環穩定性分析上二氧化矽沉析量較多之複合微米水膠因過氧化氫酶被固定地較牢固，不易變形失活而有較佳的循環穩定性，此外循環穩定性分析與存放穩定性分析的結果皆顯示(6s-PLL29/CAT)/ CTMOS=5%的穩定性最佳，推測是因為6s-PLL29會形成緻密網狀結構包覆過氧化氫酶，使之不易失活的緣故；熱穩定性分析結果為各複合微米水膠之過氧化氫酶的活性隨溫度上升而下降。本研究透過礦化作用在聚賴胺酸/過氧化氫酶微米膠上沉析二氧化矽，不僅製程簡單且複合微米水膠之各方面效能表現亦佳，具有大量運用在工業上之潛力。

The destinations of enzyme immobilization are to separate enzymes from products and raises the reuse rate. This study exhibited enzyme immobilization by emulsion, crosslinking reaction and silica mineralization. We synthesized linear and star-shaped poly(L-lysine) to encapsulate catalase and formed PLL/CAT microgels through emulsion. Then deposited silica on PLL/CAT to produce Silica/PLL/CAT hybrid materials. Linear poly(L-lysine) was l-PLL32 and star-shaped poly(L-lysine) were 3s-PLL28 and 6s-PLL29 confirmed by 1H NMR and GPC. The structure and conformation of (PLL/CAT)/Silica were checked by a series of examination. The leakage ratio analysis showed that catalase was immobilized in PLL/Silica firmly. Kinetic analysis indicated the Vmax of (6s-PLL29/CAT)/ CTMOS=5% was highest. Reusability revealed that the more silica on the (PLL/CAT)/Silica microgels, the better the reusability. Because SiO2 would reinforce the encapsulation of catalase, catalase could maintain its structure and activity well. Besides, reusability and storage stability showed the stability of (6s-PLL29/CAT)/ CTMOS=5% was the best. It could be attributed to the fact that 6s-PLL29 would form a dense structure to encapsulate catalase and consequently, it could maintain its conformation and activity. This study showed us a simple process to immobilize catalase in hybrid materials through electrostatic interaction, crosslinking reaction and mineralization, making microgels had the potential to be used in many aspects.

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