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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2014, Vol. 34 Issue (12): 78-83    DOI: 10.13523/j.cb.20141211
    
Establishing Pre-screening Method to Select Thymidine Phosphorylase Strains and Its Application
CHEN Bao-zhen1, LI Hong-mei1, WANG Wei-jie1, CHEN Qun-yi2
1. School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China;
2. Xiamen Yi He Li Biotech Co., Ltd., Xiamen 361007, China
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Abstract  

A suitable screening method to select thymidine phosphorylase high-product strains was quickly established, which was applied to the UV mutagenesis breeding. The results showed that adding 25mmol/L or less thymidine in the liquid medium did not affect the cell growth. The thymidine can be converted into thymine by the catalysis of thymidine phosphorylase in the bacterial and the OD290nm of product thymine is much higher than thymidine under the same concentration which can lead to significant increase of the fermentation broth. Based on the above principles, effective pre-screening method was established. Furtherly, the UV mutagenesis time was optimized as follows: initial mutagenesis time was 20s, after stopping 5min irradiate 10s, 15s or 20s to create mutant library by controlling the mortality rate about 80%. Through pre-screening and re-screening, the thymidine phosphorylase activity of EB27, EA42 could reach 1.025 U/mg wet cells and 0.916 U/mg wet cells, which was increased by 50% and 35% than that of the original strain. The high-product strains remained stable after transferred for 5 times.

Key words： Lactobacillus brevis      Pre-screening method      UV mutagenesis      Thymidine phosphorylase     
Received: 16 September 2014      Published: 25 December 2014
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Cite this article:

CHEN Bao-zhen, LI Hong-mei, WANG Wei-jie, CHEN Qun-yi. Establishing Pre-screening Method to Select Thymidine Phosphorylase Strains and Its Application. China Biotechnology, 2014, 34(12): 78-83.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20141211     OR     https://manu60.magtech.com.cn/biotech/Y2014/V34/I12/78


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