用于腈纶表面改性的Corynebacterium nitrilophilus腈水合酶的摇瓶发酵优化  

Fermentation Optimization in Shake Flasks of Corynebacterium nitrilophilus NHase Applied in Surface Modification of Polyacrylonitrile Fibers

作　　者：高慧慧[1,2] 陈晟[1,2] 吴敬[1,2] 陈坚[1,2] 

机构地区：[1]江南大学食品科学与技术国家重点实验室,无锡214122 [2]江南大学生物工程学院工业生物技术教育部重点实验室,无锡214122

出　　处：《中国生物工程杂志》2011年第8期54-60,共7页China Biotechnology

基　　金：国家"863"计划资助项目(2009AA02Z204)

摘　　要：通过毛细管效应、扫描电镜、染色实验等方法,考察了Corynebacterium nitrilophilus腈水合酶(nitrilre hydratase,NHase)在腈纶表面改性中的应用。结果表明,C.nitrilophilus腈水合酶处理后的腈纶纤维的润湿性及染料可染性分别比对照提高了43.5%和85.7%,说明该腈水合酶具有较好的腈纶纤维表面改性性能。为了提高用于腈纶表面改性的C.nitrilophilus腈水合酶的产量,采用单因素实验及正交实验在摇瓶上对碳源、氮源、诱导剂、金属离子等进行考察,获得较优的摇瓶发酵生产腈水合酶的条件:碳源采用葡萄糖,15 g/L;氮源采用酵母粉与氯化铵复配,浓度分别为3 g/L、1 g/L;诱导剂尿素的最适剂量为10 g/L;由于该腈水合酶是钴型酶,所以需在发酵过程中添加氯化钴,浓度为0.07 g/L。经过摇瓶优化,酶活由最初的16.2 U/ml提高到45.7 U/ml,提高了2.8倍。The application of polyacrylonitrile fibers surface modification by Corynebacterium nitrilophilus NHase was studied through capillary effect,scanning electron microscopy and dyeing experiments.The results indicated that C.nitrilophilus NHase was effective in surface modification of polyacrylonitrile fibers,which enhanced wettability and dyeing ability of polyacrylonitrile fibers by 43.5% and 85.7%.In order to improve the production of C.nitrilophilus NHase,Single-factor experiments and Orthogonal experiment were used to optimize the culture condition in shake flakes.The optimized results were as follows：15 g/L glucose was used as carbon source;Nitrogen was the combination of yeast extract and ammonium chloride,whose concentration was 3 g/L,1 g/L respectively;The optimal concentration of inducer urea was 10 g/L;The concentration of cofactor cobalt was 0.07 g/L.The NHase activity reached 45.7 U/ml compared with the initial activity of 16.2 U/ ml,which was 2.8 times after optimization in shake flakes.

关 键 词：C.nitrilophilus 腈水合酶 腈纶表面改性 发酵优化 

分 类 号：Q819[生物学—生物工程]