Promotion of prodigiosin production by introducing poly (A) tails into O-methyltransferase encoding gene
SUN Yang1, WANG Lijun1, YANG Taowei1, FU Weilai1, YI Ganfeng3, RAO Zhiming1,2*
1(Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China) 2(School of Biotechnology,Jiangnan University,Wuxi 214122,China) 3(Da Bei Nong Aquatic Science and Technology Group,Zhao'an 363500,China)
Abstract: Serratia marcescens can efficiently synthesize prodigiosin at 30 ℃ but lose this ability at 37 ℃. The expression of the key enzyme, O-methyltransferase (PigF), in the prodigiosin synthesis gene cluster was significantly reduced. In this study, six different poly (A/T) tails were added to the 3'end of pigF gene to improve mRNA stability of pigF gene, and the effect on the expression of O-methyltransferase and the synthesis of prodigiosin in S. marcescens was confirmed. The result showed that the transcriptional level of pigF was increased by 71% because the addition of “AAAAAA” can protect mRNA from degradation. Furthermore, it was found that the activity of PigF increased by 41.8% after adding “AAAAAA”. In shake flask fermentation of engineered strain JNB5-1/pigFFP, the titer of prodigiosin reached 6.21 g/L, which was 15.9% higher than that of the start strain JNB5-1. The results indicate that introduction of poly (A/T) tails can effectively increase the expression of PigF, thereby enhancing the synthesis of prodigiosin in S. marcescens.
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2021, Vol. 47 
