毛细管气相色谱法在产腈水解酶菌株筛选中的应用研究

Application of Capillary Gas Chromatography in Screening Nitrilase-Producing Strain

利用毛细管气相色谱法对发酵转化液中丙烯酸浓度的测定进行了系统研究,考察了pH值、萃取剂及萃取条件等因素对检测体系的影响,建立了毛细管气相色谱法分析产腈水解酶菌株筛选体系中丙烯酸浓度的分析方法.色谱条件为:30 m×0.32 mmi.d.,膜厚0.25μm,PEG-20 M固定液,氢焰检测器,熔融石英毛细管柱,柱温为程序升温,初始温度100℃,保留1 min后以70℃/min升到200℃.检测器温度为230℃,汽化室温度为210℃,载气(N2)的流速为7.6 mL/min.该方法检测丙烯酸和丙烯腈的相对标准偏差(RSD)(n=5)分别为0.79%,1.21%.平均回收率(n=5)分别为99.83%,100.38%.该方法操作简便,能准确测定转化体系中产物丙烯酸和底物丙烯腈的含量.

A method is described for the determination of acrylic acid produced by nitrilase in transformation liquid by gas chromatography.The influcence factors,such as pH,extraction agents,extraction conditions and other factors were studied in this paper.It was found that the optimal conditions for determining acrylic acid were as follows： PEG-20M capillary column（30 m×0.32 mm i.d.,0.25 μm,）,temperature programming（the initial temperature of 100 ℃,after 1 min rise to 200 ℃ by 70 ℃/min）,the detector and vaporization chamber temperature were 230 ℃ and 210 ℃,carrier（N2） flow rate was 7.6 mL/min.The relative standard deviation of this method（RSD）（n=5） were 0.79% and 1.21%.The average recoveries（n=5） were 99.83% and 100.38%.