食品科学 ›› 2011, Vol. 32 ›› Issue (21): 214-217.doi: 10.7506/spkx1002-6630-201121044

• 生物工程 • 上一篇    下一篇

混合反胶团提取α-淀粉酶

高树刚1,宋伟明2,安 红1   

  1. 1.淮海工学院化工学院 2.齐齐哈尔大学化学化工学院
  • 收稿日期:2018-06-20 修回日期:2018-06-20 出版日期:2011-11-15 发布日期:2011-11-11

Purification of α-Amylase Using CTAB/Tween-60 Mixed Reverse Micellar System

GAO Shu-gang1, SONG Wei-ming2, AN Hong1   

  1. (1. Department of Chemical Engineering, Huaihai Institute of Technology, Lianyungang 222005, China; 2. College of Chemistry and Chemical Engineering, Qiqihar University, Qiqihar 161006, China)
  • Received:2018-06-20 Revised:2018-06-20 Online:2011-11-15 Published:2011-11-11

摘要: 以十六烷基三甲基溴化铵(CTAB)和脱水山梨醇单硬脂酸酯聚氧乙烯醚(Tween-60)为混合表面活性剂溶于正丁醇-异辛烷中构成反胶团系统,萃取纯化α-淀粉酶。研究不同萃取条件下,α-淀粉酶的萃取率。其中反胶团相组成为:ρ(CTAB+Tween-60)=4g/L;n(CTAB):n(Tween-60)=2.0:1.0;V(正丁醇):V(异辛烷)=1.0:1.0。水相组成为:α-淀粉酶配制的粗酶液,此时c(NaCl)=0.04mol/L,水相pH11.04。结果表明:萃取温度40℃、V(有机相):V(水相) =2.0:1.0、振荡时间10min时,α-淀粉酶萃取率可达91%;反萃取水相组成为c(NaCl)=2.5mol/L、pH4.5、V(水相):V(有机相)=1.0:2.0,反萃取振荡时间10min、温度50℃时,α-淀粉酶反萃取率可达65%。反胶团相可重复使用,当V(水相):V(有机相)=1.0:2.75时,反胶团第2次α-淀粉酶萃取率达到71%。

关键词: 十六烷基三甲基溴化铵(CTAB), 吐温60(Tween60), 反胶团, α-淀粉酶, 萃取

Abstract: Reverse micellar system consisting of the surfactants cetyltrimethyl ammonium bromide (CTAB) and Tween-60 in dissolved in n-butanol-isooctane mixture was used for α-amylase purification by reversed micellar extraction. Extraction efficiency of α-amylase was investigated under different extraction conditions using a reverse micellar phase consisting of 4 g/L (CTAB +Tween-60, 2.0:1.0, n/n) in n-butanol-isooctane mixture (1.0:1.0, V/V) and an aqueous phase made up of crude α-amylase solution containing 0.04 mol/L NaCl. The extraction efficiency of α-amylase was 91% when the extraction temperature, ratio of organic phase to aqueous phase and oscillation time were 40 ℃, 2.0:1.0 and 10 min, respectively. The reverse extraction efficiency of α-amylase was 65% after 10 min oscillation at 50 ℃ when the aqueous phase was comprised of aqueous phase and organic phase at a ratio of 1.0:2.0 (V/V) at pH 4.5 containing 2.5 mol/L NaCl. The reverse micelles could be repeatedly used and the extraction efficiency of α-amylase at an aqueous phase-to-organic phase ratio of 1.0:2.75 remained 71% in the second use.

Key words: cetyltrimethylammonium bromide (CTAB), Tween-60, reverse micelle, α-amylase, extraction

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