食品科学 ›› 2010, Vol. 31 ›› Issue (15): 50-54.doi: 10.7506/spkx1002-6630-201015011

• 基础研究 • 上一篇    下一篇

方波脉冲电穿孔法提高酵母菌细胞通透性的条件优化

冀照君,孙 波* ,迟玉杰,徐 宁   

  1. 东北农业大学食品学院
  • 收稿日期:2009-11-24 修回日期:2010-06-08 出版日期:2010-08-15 发布日期:2010-12-29
  • 通讯作者: 孙 波 E-mail:bosun1962@163.com
  • 基金资助:

    黑龙江省“十一五”科技攻关计划项目(GA06B402-86-6)

Optimization of Square-wave Electroporation for Increasing the Permeability of Yeast Cells

JI Zhao-jun,SUN Bo*,CHI Yu-jie,XU Ning   

  1. College of Food Science, Northeast Agricultural University, Harbin 150030, China
  • Received:2009-11-24 Revised:2010-06-08 Online:2010-08-15 Published:2010-12-29
  • Contact: SUN Bo E-mail:bosun1962@163.com

摘要:

为了采用电穿孔法提高酵母菌细胞的通透性,将通过离心得到的酵母菌制成1 × 108CFU/mL 的菌悬液,采用不同的方波电脉冲条件对其进行处理。结果表明:在0.6mol/L 的蔗糖等渗电脉冲介质中,当酵母菌细胞的处理条件为电场强度7kV/cm,脉冲宽度50μs,脉冲数14,间隔时间2s 时,细胞的存活率平均为81.16%,电穿孔率平均为65.14%。这表明电穿孔法可以有效增强酵母菌细胞的通透性。

关键词: 方波脉冲, 电穿孔法, 酵母菌, 通透性

Abstract:

Yeast (Saccharomyces cerevisiae CICC 1012) cells were statically cultured in YEPD medium at 30 ℃ for 24 h and then separated by centrifugation at 3000 r/min after the end of culture and suspended in an electric pulse medium (containing sucrose, Tris-HCl and MgCl2, at pH 7.5) at a cell concentration of 1×108 CFU/mL before square-wave electric pulse treatment under different operating conditions. Electric field strength of 7 kV/cm, pulse duration of 50μs, pulse number of 14, time interval of 2 s, and sucrose concentration of 0.6 mol/L were found optimum. The percentage survival of yeast cells was 81.16% and the percentage of electroporated yeast cells was 65.14% under these optimum conditions. This study suggests that the permeability of yeast cells can be increased by square-wave electroporation.

Key words: square-wave electric pulse, electroporation, yeast, permeability

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