Translated Titles

Purification and Characterization of α-acetolactate Decarboxylase from Bacillus subtilis


宋麗麗(Li-Li Song);闞振榮(Zhen-Rong Kan)

Key Words

α-乙酰乳酸脫羧酶 ; 純化 ; 酶學性質 ; α-acetolactate decarboxylase ; purification ; enzymatic properties



Volume or Term/Year and Month of Publication

2005卷12期(2005 / 12 / 20)

Page #

27 - 30

Content Language


Chinese Abstract

對枯草芽孢桿菌α-乙酰乳酸脫羧酶採用硫酸銨分級沉澱、熱處理、DEAE-Sepharose Fast Flow離子交換層析進行純化,經SDS-PAGE鑒定為單蛋白帶,酶蛋白比活提高了15.5,回收率為20.3%。對酶學性質分析表明,分子量約為32kDa的單亞基酶;對熱(40℃)敏感;最適pH值和穩定pH值均為7;以α-乙酰乳酸為底物時,α-乙酰乳酸脫羧酶的動力參數Km為232.6mmol/L幾和Vmax為7.1μmol/(μg•min);Mg(上標 2+),Mn(上標 2+),Zn(上標 2+),Fe(上標 2+),Cu(上標 2+)等二價金屬離子能活酶的活性;加入EDTA後,酶活降低,用原子吸收分光光度計檢測,發現只含有Zn(上標 2+)(含量97.6nmol/mg蛋白)。

English Abstract

α-acetolactate decarboxylase from Bacillus subtilis was purified to homogeneity from cell extracts by ammonium sulfate- fractionation, heat treatment and DEAE-Sepharose Fast Flow column chromatography. The enzyme protein activity increased 15.5 and recycling rate was 20.3%. The molecular mass of this enzyme was about 32kDa. Its optimal activity was pH 7.0 and it was stable at pH 7.0 and temperature below 40℃. Kinetics parameter of α-acetolactate decarboxylase was Km=232.6mmol/l and Vmax=7.1μmo1/(μg min). Mg(superscript 2+), Mn(superscript 2+), Zn(superscript 2+), Fe(superscript 2+) and Cu(superscript 2+) had enabled effect on enzyme activity. The accession of EDTA could reduce the enzyme activity, and only Zn(superscript 2+) could be detected by atomic absorption spectrophotometer.

Topic Category 工程學 > 化學工業