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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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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.
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Received: 14 March 2016
Published: 25 September 2016
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