利用PAA進行生物矽化包覆Rhodosporidium toruloides D型胺基酸氧化酶以增進其穩定性

Stabilization of D-amino acid oxidase from Rhodosporidium toruloides by encapsulation in polyallylamine-mediated biomimetic silica

吳建成(Jian-cheng Wu)

指導教授 ： 游吉陽

大同大學/工程學院/生物工程學系所/碩士(2010年)

摘要

將RtDAO (D-Amino acid oxidase Rhodosporidium toruloides )和magnetic nanoparticle（MNP）同時包覆在polyallylamine hydrochlorid（PAA）經由生物矽化反應所形成的氧化矽中。包覆量為每克的氧化矽可包覆100 mg RtDAO（10%（w/w））；平均包覆效率為74%。包覆後RtDAO其Tm由液相之44.3℃增加至57.7℃，包覆後RtDAO之熱穩定性明顯提升。於50 mM的H2O2耐受性檢測中，固定相RtDAO其半衰期由液相之78 min增加為148 min。使用生物矽化法包覆酵素及MNP，不僅反應流程簡單而且僅需數分鐘即可完成反應，此外包覆後的酵素也具有重複使用性。因此使用生物矽化包覆DAO具有許多潛在的利用性。

關鍵字

酵素固定化 ； D型胺基酸氧化酉每 ； 生物矽化

並列摘要

D-Amino acid oxidase from Rhodosporidium toruloides (RtDAO) and Fe3O4 magnetic nanoparticles（MNP）, were encapsulated simultaneously within biomimetic silica, which was mediated by polyallylamine hydrochlor（PAA）. The encapsulation capacity for this enzyme was greater than 101 mg per gram of biomimetic silica; the average encapsulation efficiency was approximately 74%. The Tm value was increased from 44.3℃ of the free RtDAO to 57.7℃, clearly indicating the thermal stability was improved by encapsulation. In the presence of 50 mM hydrogen peroxide, encapsulated RtDAO had a half-life of 148 min, an approximately 2-fold increase in resistance to hydrogen peroxide as compared to 78-min half-life of the free form. The encapsulation process is simple and can be completed within minutes; besides, the resultant enzymes can be recovered easily under magnetic field. Such preparation of encapsulated D-amino acid oxidase could be exploited for many potential applications.

並列關鍵字

D-Amino acid oxidase ； Biomimetic silica ； Immobilized enzymes

參考文獻

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