Studies of the Trehalose-Producing Related Enzymes and the Production of Trehalose from Starch (II)

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簡歷

基本資料

Project title

Code/計畫編號

NSC92-2313-B019-034

Translated Name/計畫中文名

海藻糖生成相關酵素與海藻糖生產之研究(2/3)

Project Coordinator/計畫主持人

Tsuei-Yun Fang

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Food Science

Website

https://www.grb.gov.tw/search/planDetail?id=850955

Year

2003

Start date/計畫起

01-08-2003

Expected Completion/計畫迄

31-07-2004

Bugetid/研究經費

1051千元

ResearchField/研究領域

食品科技(農)

在第一年度計畫中，已利用聚合脢鏈反應，並配合其他基因重組技術，將Sulfolobus solfataricus的TreX、TreY 及TreZ 三種基因成功選殖並表現於大腸桿菌中。我們部分純化了肝醣枝切脢(GDE)與海藻糖甘糊精生成脢，也探討兩酵素的一些特性;已知GDE 的酵素最適作用溫度約為75℃，最適作用pH 為 5; TDFE 的酵素最適作用溫度約為65 至75℃，最適作用pH為 5。在本(第二)年度計畫中，我們繼續GDE、TDFE 與TFE 的純化，並進行酵素特性探討。利用熱處理、離子交換層析與膠體過濾層析已獲得純化之GDE。純化後之GDE，其酵素最適作用溫度與pH 仍為75℃及pH5。GDE 在40-80 的溫度下保甘兩小時仍有良好之酵素活性，在pH 值5-10 的範圍，具有良好的pH 穩定性。許多金屬離子，例如Ca2+、Cu2+等，對GDE 具有提高活性的效果。利用熱處理、超過濾與膠體過濾層析，已獲得純化之TDFE。純化後之TDFE，其酵素最適作用溫度與pH 仍為75℃及pH5。TDFE 在45-80 的溫度下保甘兩小時仍有良好之酵素活性，在pH 值4.5-11 的範圍，具有良好的pH 穩定性。金屬離子，例如 Cu2+、Zn2+等，對TDFE 具有抑制活性的效果。利用熱處理與離子交換層析已獲得純化之TFE。純化後之TFE，其酵素最適作用溫度為65-70℃ In the first year of this proposal, we had successfully cloned TreX, TreY, and TreZ genes from Sulfolobus solfataricus ATCC35092 and expressed recombinant GDE, TDFE and TFE in E. coli BL21 (DE3)-CodonPlus-RIL, respectively. We had partially purified and characterized GDE and TDFE. The partially purified GDE exhibited optimum activity at 75.degree.C and pH 5. The partially purified TDFE exhibited optimum activity at 65 ~ 75.degree.C and pH 5. In the second year of this proposal, we continued the purification and characterization of GDE, TDFE and TFE. The GDE was purified sequentially by using heat treatment, ion exchange, and gel filtration. The purified GDE showed an apparent optimal pH of 5 and an optimal temperature at 75.degree.C. The enzyme was stable at pHs ranging from 5 to 10, and the activity remained unchanged after a 2-h incubation at 80.degree.C. Metal ions, such as Ca2+ and Cu2+, can enhance the activity of GDE. The TDFE was purified sequentially by using heat treatment, ultrafiltration, and gel filtration. The purified TDFE showed an apparent optimal pH of 5 and an optimal temperature at 75.degree.C. The enzyme was stable at pHs ranging from 4.5 to 11, and the activity remained unchanged after a 2-h incubation at 80.degree.C. Metal ions, such as Cu2+ and Zn2+, can inhibit the activity of GDE. The TFE was purified sequentially by using heat treatment and ion exchange chromatography. The obtained GDE showed an apparent optimal temperature at 65-70.degree.C.

Keyword(s)

Trehalose
Glycogen debranching enzyme
Trehalosyl dextrins forming enzyme
Trehalose forming enzyme
Sulfolobus solfataricus

DSpace CRIS

Studies of the Trehalose-Producing Related Enzymes and the Production of Trehalose from Starch (II)

基本資料

Description