食品与发酵工业 >

2021 , Vol. 47 >Issue 9: 49 - 56

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.025669

研究报告

氧甲基转移酶编码基因ploy (A)加尾促进粘质沙雷氏菌合成灵菌红素能力研究

  • 孙杨 ,
  • 王丽君 ,
  • 杨套伟 ,
  • 付维来 ,
  • 易敢峰 ,
  • 饶志明
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  • 1(工业生物技术教育部重点实验室(江南大学),江苏 无锡,214122)
    2(江南大学 生物工程学院,江苏 无锡,214122)
    3(大北农水产科技集团,福建 诏安,363500)
博士研究生(饶志明教授为通讯作者,E-mail:raozhm@jiangnan.edu.cn)

收稿日期: 2020-09-15

  修回日期: 2020-10-20

  网络出版日期: 2021-06-03

基金资助

国家自然科学基金项目(31870066;21778024)

收起

Promotion of prodigiosin production by introducing poly (A) tails into O-methyltransferase encoding gene

  • SUN Yang ,
  • WANG Lijun ,
  • YANG Taowei ,
  • FU Weilai ,
  • YI Ganfeng ,
  • RAO Zhiming
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  • 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)

Received date: 2020-09-15

  Revised date: 2020-10-20

  Online published: 2021-06-03

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摘要

粘质沙雷氏菌能在30 ℃ 高效合成灵菌红素,但在37 ℃甚至更高温度则丧失合成能力。研究表明,在37 ℃甚至更高温度下,粘质沙雷氏菌中灵菌红素合成关键酶氧甲基转移酶(O-methyltransferase,PigF)的表达量显著下降。因此,通过人工在pigF基因3′端添加6条不同的poly(A/T)尾来提高pigF基因mRNA稳定性,并观察其对粘质沙雷氏菌中PigF表达量及灵菌红素合成的影响。结果发现,添加“AAAAAA”可以保护mRNA免受降解,从而使得pigF转录水平提高71%。其次,添加“AAAAAA”后,PigF比酶活力提高了41.8%。最后通过摇瓶发酵,实验结果表明,与30 ℃培养的原始菌株相比,工程改造的JNB5-1/pigFFP菌株的灵菌红素产量达到6.21 g/L,提高了15.9%。该研究表明,通过引入poly(A/T)尾提高了PigF的表达量,进而提高了粘质沙雷氏菌合成灵菌红素的能力。

关键词： 粘质沙雷氏菌; 灵菌红素; 氧甲基转移酶(PigF); 温度

本文引用格式

孙杨 , 王丽君 , 杨套伟 , 付维来 , 易敢峰 , 饶志明 . 氧甲基转移酶编码基因ploy (A)加尾促进粘质沙雷氏菌合成灵菌红素能力研究[J]. 食品与发酵工业, 2021 , 47(9) : 49 -56 . DOI: 10.13995/j.cnki.11-1802/ts.025669

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.

Key words： Serratia marcescens; prodigiosin; O-methyltransferase (PigF); polyadenylation; temperature

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