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| A Rapid Method of Screening for Thermostable Transglutaminase from Streptomyces mobaraensis |
| GUO Wei-ting1, ZHANG Hui1, ZHA Dong-feng2, HUANG Han-feng2, HUANG Jing1, GAO Hong-liang1, CHANG Zhong-yi1, JIN Ming-fei1, LU Wei3 |
1. School of Life Science, East China Normal University, Shanghai 200241, China;
2. Economic Crops Center Henan Shangqiu Agriculture Bureau, Shangqiu 476000, China;
3. Taixing Dongsheng Food and Technology Limited Company, Taixing 225411, China |
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Abstract Objective:The aims are to obtain thermostable MTG from Streptomyces mobaraensis mutants library producing by nitrosoguanidine (NTG) radom mutagenesis and high-throughput screening.Methods:A rapid method of screening for thermostable MTG from Streptomyces mobaraensis is established by optimization of high-throughput MTG activity assay, screening temperature and time, respectively.Streptomyces mobaraensis mutants library is obtained by optimization of the concentration of NTG. A mutant 12-82 with thermostable MTG was chosen by preliminary screening and second screening from the 5200 mutants library. Results:The spores from Streptomyces mobaraensis were treated by NTG (2mg/ml, pH8.0) for 60 min, then the fermented supernatant was incubated at 70℃ for 7.5 min after fermentation in 96 well microtiters. Finally, MTG activity was determined at 37℃ air bath for 10 min.5 mutants with thermostable MTG were selected from 5200 mutants.When incubated at 50℃ for 60min and 70℃ for 1.5min, the relative activity of 12-82 was almost 20% higher than the parent strain.When incubated at 80℃ for 2min, the relative activity of 12-82 was 11.9%. Conclusion:5 mutants with thermostable MTG were selected by the screening method of NTG random mutagenesis combined with high-throughput screening.Among them, the relative activity of the mutant 12-82 was 10%~20% higher than that of the parent strain when incubated at 50℃,70℃ and 80℃, respectively.This established rapid method to obtain thermostable MTG is practical and the increased thermostability of MTG will be advantageous for its application in food processing at high temperature.
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Received: 29 April 2015
Published: 25 August 2015
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Cite this article:
GUO Wei-ting, ZHANG Hui, ZHA Dong-feng, HUANG Han-feng, HUANG Jing, GAO Hong-liang, CHANG Zhong-yi, JIN Ming-fei, LU Wei . A Rapid Method of Screening for Thermostable Transglutaminase from Streptomyces mobaraensis. China Biotechnology, 2015, 35(8): 83-89.
URL:
https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150812 OR https://manu60.magtech.com.cn/biotech/Y2015/V35/I8/83
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