β-半乳糖甘酶之固定化與其性質之研究

The Study of Preparation and Properties of Immobilized β-galactosidase

10.29987/CCUHKJE.201206.0005

王星堯(S. Y. Wang)；吳誌明(J. M. Wu)

固定化酵素 ； 半乳糖甘 ； 細胞通透 ； 酵母菌 ； 洋菜膠 ； SDS/chloroform ； immobilized ； galactosidase ； permeability ； yeast ； agar ； SDS / chloroform

華岡工程學報

29期（2012 / 06 / 01）

23 - 28

繁體中文

本研究使用能表現半乳糖甘酶β-galactosidase（β-gal）之Pichia Pastoris酵母菌，使用SDS/chloroform進行Pichia Pastoris酵母菌細胞通透（permeabilization），再利用洋菜膠（agar）將其進行固定。實驗分別以5種試劑進行酵母菌細胞通透測試，結果顯示以01.94×10^-3 ％ SDS /3.80 ％ chloroform通透效果最佳，其β-gal活性可達7.5×10^3 unit/mg cell，在將通透過後的酵母菌細胞用洋菜膠進行固定化測試，結果顯示以2% 洋菜膠之固定化效果最好。對此固定化酵素進行各種反應條件探討，最佳條件如下：在pH＝7.5，溫度28℃，反應20分鐘的情況下酵素活性有最高值3.4×10^2 unit/mg cell。

β-D-galactosidase from Pichia pastoris ZGZA.Permeabilization of Pichia pastoris cells in relation to β-gal activity was carried out using SDS/chloroform to avoid the problem of β-gal activity. The cell Permeabilization were tested by using different ratio of SDS/chloroform and get a best ratio (1.94 × 10-3 % SDS / 3.80% of chloroform) to give highest β-gal activity (7.5 × 103 unit / mg cell).Next, the cell was immobilized on agar. Different process parameters (biomass load, pH, temperature, and incubation time) were optimized to enhance the highest β-gal activity .The highest β-gal activity was observed Immobilized of 2% agar with 9.94 mg DW yeast biomass after 20 min of incubation period at optimum pH of 7.5and temperature of 28℃.