天津科技大学学报2018，Vol.33Issue(2)：20-26,7.DOI:10.13364/j.issn.1672-6510.20160416
常压室温等离子体(ARTP)诱变及高通量筛选维生素B12高产菌株
Mutation by Using Atmospheric Pressure and Room Temperature Plasmas and High-throughput Screening Method for Improving Vitamin B12 Production
摘要
Abstract
Atmospheric pressure and room temperature plasma (ARTP) mutagenesis system was used to induce Pseudomonas denitrificans.In order to obtain mutant strains with enhanced ability to produce VB12,the optimum conditions for mutation were determined,which were 75 s as the mutagenesis time and 100 W as the output power.A method was constructed with flow cytometry and riboswitch to efficiently identify positive mutant strains which were cultivated with 48-well microplates and detected by microplate reader as high-yielding mutants.Thus,a screening system for vitamin B12 high throughput was built.Through four rounds of ARTP mutation and high-throughput screening,PA320-M4-1B1 mutants (103.2 ± 2.1)mg/L were obtained,which increased the contents of VB12 by 43.8％ compared with the control strain PA320 (71.9 ± 1.8)mg/L,when the strains were cultivated on a rotary shaker at 30 ℃ for 6 d.关键词
常压室温等离子体(ARTP)/维生素B12/核糖开关/高通量筛选Key words
ARTP/vitamin B12/riboswitch/high-throughput screening分类
生物科学引用本文复制引用
蔡莹瀛,夏苗苗,董会娜,张同存,张大伟..常压室温等离子体(ARTP)诱变及高通量筛选维生素B12高产菌株[J].天津科技大学学报,2018,33(2):20-26,7.基金项目
天津市自然科学基金资助项目(16JCYBJC23500) （16JCYBJC23500）
天津市杰出青年科学基金资助项目(17JCJQJC45300) （17JCJQJC45300）
