四氢嘧啶吸收缺陷突变株高效制备四氢嘧啶

【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 salina DSM 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。

Construction of ectoine absorption defective mutant for efficient ectoine production

[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 5928T(teaABC-) by Red recombination technology.[Results] The total concentration of ectoine and productivity in a 10 L fermentor 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.