亲和反胶团选择性萃取分离酵母乙醇脱氢酶

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亲和反胶团选择性萃取分离酵母乙醇脱氢酶

张天喜; 刘会洲; 陈家镛

Laboratory of Separation Science and Engineering, Institute of Chemical Metallurgy Chinese
Academy of Sciences, Beijing 100080, China

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2001-09-28 发布日期:2001-09-28
通讯作者: 张天喜

Selective Affinity Separation of Yeast Alcohol Dehydrogenase by Reverse Micelles with
Unbound Triazine Dye

ZHANG Tianxi; LIU Huizhou; CHEN Jiayong

Laboratory of Separation Science and Engineering, Institute of Chemical Metallurgy Chinese
Academy of Sciences, Beijing 100080, China

Received:1900-01-01 Revised:1900-01-01 Online:2001-09-28 Published:2001-09-28
Contact: ZHANG Tianxi

摘要/Abstract

摘要： The reversed micelles were formed with cationic cetyltrimethylammonium bromide (CTAB) as
surfac tant and n-hexanol as cosolvent in the CTAB (50mmol.L-1)/hexanol (15% by
volume)/hexane system. Cibacron Blue 3GA (CB) as an affinity ligand in the aqueous phase
was directly introduced to the reversed micelles with electrostatic interaction between
anionic CB and cationic surfactant. High molecular weight (Mr) protein, yeast alcohol
dehydrogenase (YADH, Mr = 141000) from baker’s yeast, has been purified using the affinity
reversed micelles by the phase transfer method. Various parameters, such as CB
concentration, pH and ionic strength, on YADH forward and backward transfer were studied.
YADH can be transferred into and out from the reversed mi celles under mild conditions
(only by regulation of solution pH and salt concentration) with the successful recovery of
most YADH activity. Both forward and backward extractions occurred when the aqueous phase
pH＞pI with electrostatic attraction between YADH and CTAB. The recovery of YADH activity
and purification factor have been improved with addition of a small amount of affinity CB.
The recovery of YADH activity obtained was ～99% and the purification factor was about 4.0
-fold after one cycle of full forward and backward extraction. The low ionic strength in
the initial aqueous phase might be responsible for the YADH transfer into the reversed
micellar phase.

关键词: reversed micelles;yeast alcohol dehydrogenase;protein purification;affinity technology; cetyltrimethy lammonium bromide

Abstract: The reversed micelles were formed with cationic cetyltrimethylammonium bromide (CTAB) as
surfac tant and n-hexanol as cosolvent in the CTAB (50mmol.L-1)/hexanol (15% by
volume)/hexane system. Cibacron Blue 3GA (CB) as an affinity ligand in the aqueous phase
was directly introduced to the reversed micelles with electrostatic interaction between
anionic CB and cationic surfactant. High molecular weight (Mr) protein, yeast alcohol
dehydrogenase (YADH, Mr = 141000) from baker’s yeast, has been purified using the affinity
reversed micelles by the phase transfer method. Various parameters, such as CB
concentration, pH and ionic strength, on YADH forward and backward transfer were studied.
YADH can be transferred into and out from the reversed mi celles under mild conditions
(only by regulation of solution pH and salt concentration) with the successful recovery of
most YADH activity. Both forward and backward extractions occurred when the aqueous phase
pH＞pI with electrostatic attraction between YADH and CTAB. The recovery of YADH activity
and purification factor have been improved with addition of a small amount of affinity CB.
The recovery of YADH activity obtained was ～99% and the purification factor was about 4.0
-fold after one cycle of full forward and backward extraction. The low ionic strength in
the initial aqueous phase might be responsible for the YADH transfer into the reversed
micellar phase.

Key words: reversed micelles, yeast alcohol dehydrogenase, protein purification, affinity technology, cetyltrimethy lammonium bromide

张天喜, 刘会洲, 陈家镛. 亲和反胶团选择性萃取分离酵母乙醇脱氢酶[J]. CIESC Journal.

ZHANG Tianxi, LIU Huizhou, CHEN Jiayong. Selective Affinity Separation of Yeast Alcohol Dehydrogenase by Reverse Micelles with
Unbound Triazine Dye[J]. .

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亲和反胶团选择性萃取分离酵母乙醇脱氢酶

Selective Affinity Separation of Yeast Alcohol Dehydrogenase by Reverse Micelles with
Unbound Triazine Dye

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亲和反胶团选择性萃取分离酵母乙醇脱氢酶

Selective Affinity Separation of Yeast Alcohol Dehydrogenase by Reverse Micelles with Unbound Triazine Dye

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Selective Affinity Separation of Yeast Alcohol Dehydrogenase by Reverse Micelles with
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