摘要
为获得可有效降解植物细胞壁中纤维素的高产纤维素酶菌株,将具有安全可靠、不产生毒素、生长速度快且发酵周期短等优势的黑曲霉作为出发菌株。利用重离子^(12)C^(6+)束流按照0,40,70,100,130,160,190 Gy的辐照剂量对黑曲霉原始菌株进行辐照,经平板初筛后得到两株优良菌株;然后添加α-纤维素进行了8周自适应性进化,通过连续5代发酵该菌株生长性能稳定;将得到的每代菌株进行胞内蛋白分子量的对比,得到一株最优菌株CJH-JWSFZh-W122,将其摇瓶发酵测定纤维素酶活性,其产纤维素酶的滤纸酶活(FPA)到了223.5 U/mL,内切葡聚糖酶活(CMC)达到了440.8 U/mL,分别比原始菌株提高了6.07%和8.01%。突变株在连续5代发酵后仍保持了良好的发酵性能。对突变菌株CJH-JWSFZh-W122和原始菌株进行了RAPD分析和SDS-PAGE电泳分析,证实了该菌株存在基因突变。
In order to obtain high-yielding cellulase strains that can effectively degrade cellulose in plant cell walls,Aspergillus niger,which has the advantages of safety and reliability,non-toxicity,fast growth and short fermentation period,was used as the starting strain.The original strain,Aspergillus niger,was irradiated by heavy ion beam^(12)C^(6+)(irradiation doses:40,70,100,130,160 and 190 Gy),and two excellent strains were obtained after primary screening on the plate;8 weeks of self-adaptive evolution,the strain has stable growth performance through 5 consecutive generations of fermentation;the obtained strains of each generation were compared for the molecular weight of intracellular proteins,and an optimal mutant strain CJHJWSFZh-W122 was obtained,which was determined by shaking flask fermentation Cellulase activity,the filter paper activity(FPA)of cellulase production reached 223.5 U/mL,and the carboxymethyl cellulase activity(CMCase)reached 440.8 U/mL,which were 6.07%and 8.01%higher than the original strains,respectively.The excellent fermentation performance was still maintained after 5 consecutive generations;RAPD analysis and SDS-PAGE electrophoresis analysis were applied to compare the mutant strain CJH-JWSFZh-W122 and original strain,which confirmed the gene mutation of this strain.
作者
刘浩
陈皓
周翔
田雪娇
马良
胡伟
闫永利
韩玲
LIU Hao;CHEN Hao;ZHOU Xiang;TIAN Xuejiao;MA Liang;HU Wei;YAN Yongli;HAN Ling(College of Food Science and Engineering,Gansu Agricultural University,Lanzhou 730070,China;Institute of Modem Physics,Chinese Academy of Sciences,Lanzhou 730000,China;University of Chinese Academy of Sciences,Beijing 100049,China)
出处
《原子核物理评论》
CAS
CSCD
北大核心
2022年第3期382-390,共9页
Nuclear Physics Review
基金
内蒙古科技重大专项子课题项目(E039211SZ1)
甘肃省科技计划项目(20YF3NA016)
关键词
黑曲霉
纤维素酶
重离子束
Α-纤维素
适应性进化
Aspergillus niger
cellulase
heavy ion beam
alpha-cellulose
adaptive evolution
