文献类型：期刊文献

中文题名：N^+注入诱变选育纤维素酶高产菌株及发酵产酶营养因子优化研究

英文题名：Screening of High-producing Cellulase Strain by Low Energy N^+ Implantation and Optimization of Fermentation Nutrient Factors

作者：张宁[1,2,3,4] 蒋剑春[1,2,3,4] 杨静[1,2,3,4] 卫民[1,2,3,4] 赵剑[1,2,3,4] 童娅娟[1,2,3,4]

第一作者：张宁

机构：[1]中国林业科学研究院林产化学工业研究所;[2]生物质化学利用国家工程实验室;[3]国家林业局林产化学工程重点开放性实验室;[4]江苏省生物质能源与材料重点实验室

年份：2014

卷号：48

期号：4

起止页码：28-34

中文期刊名：生物质化学工程

外文期刊名：Biomass Chemical Engineering

基金：国家自然科学基金资助项目(31100429)

语种：中文

中文关键词：N+离子注入;里氏木霉;纤维素酶

外文关键词：N＋ion implantation; Trichoderma reesei; cellulase

分类号：TQ352;Q815

摘要：应用低能氮离子(N+)注入技术对纤维素酶产生菌里氏木霉(Trichoderma reesei)进行诱变选育,在能量为10 keV,注量为150×1014和200×1014N+/cm2的条件下分别筛选得到3株纤维素酶高产菌株,连续5代遗传稳定性实验结果表明,所得到的高产菌株遗传稳定性较好,羧甲基纤维素酶活力均提高到3.300 IU/mL以上,较出发菌株(2.698 IU/mL)提高了20.0%以上。采用Plackett-Burman实验设计法和旋转中心组合设计法系统地研究高产菌株150-1-1发酵营养因子组成,得到了纤维素酶产量随葡萄糖、麸皮和微晶纤维素等营养因子的变化规律及相应的响应面分析图。实验结果表明,葡萄糖、麸皮和微晶纤维素浓度与纤维素酶活存在显著的相关性,当葡萄糖浓度为4.9 g/L,麸皮浓度为23.0 g/L,微晶纤维素浓度为7.7 g/L时,150-1-1纤维素酶滤纸酶活力达到2.439 IU/mL,较优化前(2.000 IU/mL)提高了22.0%。
Cellulase producing strain Trichoderma reesei was mutated by low energy N＋implantation. When the implantation energy was 10 keV,and the implantation doses were 150 × 1014and 200 × 1014N＋/cm2,three high yield mutant strains were obtained. Their cellulase yield reached 3. 300 IU /mL,and improved over 20. 0% than that of original strain（ 2. 698 IU /mL）.Furthermore,mutant strains had better heredity stability after five passages. Plackett-Burman and central composite rotatable design experiment were applied to optimize the concentration of nutrient factors for cellulase production. The changing patterns of glucose,wheat bran and microcrystalline cellulose were obtained,and the corresponding response surface analysis diagram were also obtained. Experimental results showed that glucose,wheat bran and microcrystalline cellulose had an individual significant influence on cellulose production. The optimum concentration of nutrient factors for cellulase production was 4. 9 g /L of glucose,23. 0 g /L of wheat bran,and 7. 7 g /L of microcrystalline cellulose. Under the optimized conditions,the cellulase yield of T. reesei150-1-1 reached 2. 439 IU /mL,with a 22% increase than that before optimization.