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一株产腈水合酶菌株酶活高效表达条件的研究
引用本文:李志东,李娜,张洪林,王战勇,邱峰.一株产腈水合酶菌株酶活高效表达条件的研究[J].精细化工,2006,23(12):1201-1204,1210.
作者姓名:李志东  李娜  张洪林  王战勇  邱峰
作者单位:辽宁石油化工大学,环境工程系,辽宁,抚顺,113001
摘    要:以抚顺腈纶厂废水中筛选的一株Nocardia sp为出发菌株,通过逐渐增加培养基中丙烯腈的体积浓度重复继代培养,得到1株酶活高效表达的Nocardia sp菌,酶活比出发菌株提高了15倍。确定了菌株培养的最优条件:葡萄糖(25 g/L),诱导剂脲(0.06 g/L)、Co2+(0.02 g/L)、丙烯酰胺(10%),反应条件pH(7)、温度(20℃),底物丙烯腈体积分数(<5%),产物丙烯酰胺的质量分数(<20%)。正交实验表明,反应温度和脲的加入量为反应过程中的显著因素。

关 键 词:腈水合酶  诺卡氏菌  驯化选育  产酶条件
文章编号:1003-5214(2006)12-1201-05
收稿时间:2006-05-10
修稿时间:2006-05-102006-07-17

Study of the Conditions for a Nitrile Hydratase Producing Strain to Produce Enzyme Efficiently
LI Zhi-dong,LI Na,ZHANG Hong-lin,WANG Zhan-yong,QIU Feng.Study of the Conditions for a Nitrile Hydratase Producing Strain to Produce Enzyme Efficiently[J].Fine Chemicals,2006,23(12):1201-1204,1210.
Authors:LI Zhi-dong  LI Na  ZHANG Hong-lin  WANG Zhan-yong  QIU Feng
Affiliation:Department of Environment Engineering, Liaoning University of Petroleum and Chemical Technology, Fushun 113001, Liaoning , China
Abstract:A bacterial strain of Nocardia sp which was filtered from waste water of Fushun Acrylonitrile Factory having nitrile hydratase activity and acrylonitrile tolerance was developed by repeated subculturing in the broth containing acrylonitrile with successively slightly increased acrylonitrile concentration.Enzyme activity of the domesticated strain was 15 times higher than that of original Nocardia sp.The optimum conditions of cultivation were glucose 25 g/L,urea 0.06 g/L,acrylonitrile<5%,acrylamide<20%,Co~(2+) 0.02 g/L,pH=7.0 and temperature 22 ℃.An orthogonal experiment of L_9(3~4)was devised,which can make certain that temperature and urea are the prominent factors.
Keywords:nitrile hydratase  Nocardia sp  domestication  nitrile hydratase producing conditions
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