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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2016, Vol. 36 Issue (9): 47-53    DOI: 10.13523/j.cb.20160906
技术与方法     
常压室温等离子体(ARTP)诱变茂源链霉菌菌株
田淑翠1, 牛延宁1, 常忠义1, 高红亮1, 步国健2, 金明飞1
1 华东师范大学生命科学学院 上海 200241;
2 泰兴市东圣食品科技有限公司 泰兴 225411
The Streptoverticillium mobaraense Mutagenesis Using Atmospheric Pressure Plasma at Room Temperature (ARTP) Method
TIAN Shu-cui1, NIU Yan-ning1, CHANG Zhong-yi1, GAO Hong-liang1, BU Guo-jian2, JIN Ming-fei1
1 Life Science College of East China Normal University, Shanghai 200241, China;
2 Taixing Dongsheng Food Science and Technology Limited Company, Taixing 225411, China
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摘要：

采用常压室温等离子体（ARTP）诱变技术处理茂源链霉菌（Streptoverticillium mobaraense）菌株HS47的孢子，选育微生物谷氨酰胺转胺酶（MTG）高产菌株。菌株的致死率强度和正突变率高低结果表明，在电源功率为120W，处理距离2mm，工作气流量10slpm时，等离子体氦气对茂源链霉菌HS47孢子的最佳处理时间为30s。将诱变后的孢子液稀释涂布后，利用96孔板高通量筛选方法对单菌落进行初筛，选出高产的突变株进行两轮试管复筛，筛选过程中保持对菌株的分离纯化，最终获得一株高产菌株M-8，其MTG酶活由2.8U/ml提高到5.1U/ml，较出发菌株HS47提高了82%。该菌株的摇瓶发酵实验证明，其酶活的提高是单位菌株分泌的MTG有所增加的结果。经过8次传代，证实该菌株具有良好的遗传稳定性。这为谷氨酰胺转胺酶的工业化生产提供了菌种支持和理论支持。
关键词： 常温室压等离子体诱变;  高通量筛选;  茂源链霉菌;  谷氨酰胺转胺酶    
Abstract:

To get microorganism transglutaminase (MTG) high yield strains, the spores of Streptoverticillium mobaraense strain HS47 were mutagenized using atmospheric pressure plasma at room temperature (ARTP) method.The fatality rate strengthand positive mutation rate result of the strains reveal that when the power is 120W, handling distance is 2mm, working flow is 10slpm,the optimum time for the plasma heliumto mutagenized spores of S.mobaraense HS47 was 30s.Spread the mutagenic spores diluent to tablets and screen single colonies using 96-well plates high-throughput screening method. Pick out the high yield mutant strains to go on with two rounds of test tube screening. Among the process, keep the separation and purification of strains. Finally, one high yield strain M-8 was found. Its MTG enzyme activity was 5.1U/ml,increased by 82% compared to the starting strain HS47 whose MTG enzyme activity was only 2.8U/ml.The reaults of shaking flask fermentation of strain M-8 show that its increase in the enzyme activity results from the increase of MTG secretion of unit strain.The MTG enzyme activity of M-8 of eight batches have no significant difference, so the strain has good genetic stability. The found of M-8 provides strain support and theoretical support for industrialized production of MTG.
Key words: High-through screening    Streptoverticillium mobaraense    Transglutaminase    ARTP mutagenesis
收稿日期: 2016-03-14 出版日期: 2016-09-25
ZTFLH:  Q819  
基金资助:

中央高校基本科研业务费专项资金资助项目（78210203）
通讯作者: 金明飞     E-mail: mfjin@bio.ecnu.edu.cn
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引用本文:

田淑翠, 牛延宁, 常忠义, 高红亮, 步国健, 金明飞. 常压室温等离子体(ARTP)诱变茂源链霉菌菌株[J]. 中国生物工程杂志, 2016, 36(9): 47-53.

TIAN Shu-cui, NIU Yan-ning, CHANG Zhong-yi, GAO Hong-liang, BU Guo-jian, JIN Ming-fei. The Streptoverticillium mobaraense Mutagenesis Using Atmospheric Pressure Plasma at Room Temperature (ARTP) Method. China Biotechnology, 2016, 36(9): 47-53.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20160906    或    https://manu60.magtech.com.cn/biotech/CN/Y2016/V36/I9/47

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