四氢嘧啶吸收缺陷突变株高效制备四氢嘧啶

摘要

[Objective] To further enhance ectoine (1, 4, 5, 6-tetrahydro-2-methyl-4- pyrimidinecarboxylic acid) synthesis efficiency. [Methods] We cloned Halomonas salina DSM 5928T teaABC gene of ectoine-specific transporter TeaABC by walking PCR and constructed ectoine absorption defective mutant H. salina DSM 5928 (teaABC) by Red recombination technology. [Results] The total concentration of ectoine and productivity in a 10 L fennentor were 9. 10 (±0.08) g/L and 9.93 (±0.09) g/L. d. [Conclusion] The ectoine absorption defect mutant H. salina DSM 5928T (teaABC-) compromised the negative feedback regulation of ectoine synthesis, which can significantly improve the efficiency of ectoine production.%[目的]为进一步提高四氢嘧啶(1,4,5,6-四氢-2-甲基-4-嘧啶羧酸；1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid；ectoine)合成效率,[方法]利用步移PCR方法克隆了Halomonas salinaDSM 5928T四氢嘧啶特异性转运蛋白(ectoine-specific transporter; TeaABC)编码基因teaABC,Red重组技术构建了四氢嘧啶吸收缺陷突变株H.salina DSM 5928T(teaABC -).[结果]H.salina DSM 5928T(teaABC -)10 L发酵罐的四氢嘧啶发酵,四氢嘧啶总浓度9.10(±0.08) g/L,合成效率为9.93(±0.09) g/L.d.[结论]四氢嘧啶吸收缺陷突变株H.salina DSM 5928T(teaABC-),解除了四氢嘧啶吸收对其合成的负反馈调节,从而显著提高了四氢嘧啶合成效率.